CN115612114B - Copolymer membrane and method for enzymatic self-assembly synthesis and application thereof - Google Patents

Abstract

The copolymer film and its enzymatic self-assembly synthesis method provided by the invention are prepared at the interface of two liquid phases through an enzymatic reaction of a film-forming monomer containing a phenolic hydroxyl group, a film-forming monomer containing at least two amino groups and a catalyst. A copolymer film was obtained. The method has simple operation steps, mild reaction conditions, and a wide selection of film-forming monomers. The film-forming monomers can be selected according to application requirements to prepare copolymer films with corresponding functions. The preparation method of the present invention can be easily upgraded from laboratory to industrial mass production, and has good application prospects.

Description

Copolymer membranes and their enzymatic self-assembly synthesis methods and applications

Technical field

The invention belongs to a new type of polymer material and belongs to the fields of modern chemistry, biology, material science and its applied science and engineering. Specifically, it relates to a type of copolymer film and its enzymatic self-assembly synthesis method and application at the liquid-liquid interface.

Background technique

There are millions of organic compounds discovered so far, but the types of monomers that can actually be used as raw materials for synthetic polymers are quite limited, with hundreds of species at most. Therefore, how to use two or more monomers in different ways and Copolymerization is carried out in different proportions to obtain a wide variety of copolymers with different properties to meet different use requirements [Wang Huaisan, Kou Xiaokang. Polymer Chemistry Tutorial [M]. Beijing: Science Press, 2007: 213] still It is an important research and development direction. The design and enzymatic synthesis of new functional polymer materials, the synthesis and application of nanopolymers are all at the forefront of polymer research and development.

Two-dimensional materials and quasi-two-dimensional materials are used in cutting-edge fields such as quantum information, artificial intelligence, integrated circuits, life and health, brain science, aerospace technology, as well as sensors, logic switches, pharmaceutical preparations, medical diagnosis, electronic information, flexible wearable devices, There are broad application prospects in the fields of smart clothing, food safety testing, environmental protection, and smart packaging. Among them, films with a thickness on the order of nanometers (1-100nm) can be called "nano-thin films" and will express the properties of nanomaterials.

Large-scale synthesis of high-quality films with broad tunability will advance the development of artificial solids with engineered functions. However, many "bottom-up" polymer membrane preparation technologies still have obvious shortcomings in aspects such as controllability. The thickness control of many film layers, especially the thickness and uniformity of nano-films, is still a difficulty in process implementation. Uncontrollable reactions make it difficult to achieve large-scale industrial production, and there are even fewer methods that can be used to prepare ultra-thin multilayer films of organic polymer composites.

The stability of the membrane is very important for the application of the membrane. LB membranes (prepared by Langmuir-Blodgett film-forming technology, monolayer or multi-layer membranes with molecules arranged in an orderly manner) are uniformly and completely regularly arranged at the molecular level. However, the mechanical strength of the LB film deposited by small molecules is not high (only a few Newtons of force can scratch the LB film deposited on the substrate), and its heat resistance, solvent resistance and environmental resistance are poor. Practical applications are subject to certain limitations. In order to overcome these weaknesses, an effective method is to introduce groups that can further undergo polymerization reactions into film-forming organic small molecule compounds to make them polymerized [He Pingsheng. Polymerization in a two-dimensional state: monomolecular films and Langmuir -Polymerization of Blodgett membrane [M]. Hefei: University of Science and Technology of China Press, 2008: 58.]. However, LB film requires that the film-forming organic compound monomer molecules be amphiphilic and have appropriate molecular chain lengths, which greatly limits the selection range of LB film-forming monomers and brings problems to the design of LB film-forming materials. difficulty. Moreover, since the polymerization of monomers into polymers is a transition from van der Waals interaction distance to chemical bond distance, the shortening of the distance between molecules leads to the shrinkage of the formed monomolecular film polymer or polymeric LB film, and the overall order of the film is lost. To ensure [He Pingsheng. Polymerization in two-dimensional state: polymerization of monomolecular membranes and Langmuir-Blodgett membranes [M]. Hefei: University of Science and Technology of China Press, 2008: 61.].

Supramolecular materials are a new type of modern material in the development stage. They are based on supramolecular chemistry and utilize non-covalent bonds between molecules (such as hydrogen bond interactions, electron donor-acceptor interactions). Materials prepared by interaction, ionic interaction, etc.) [Zhang Zhijie. Physical Chemistry of Materials [M]. Beijing: Chemical Industry Press, 2020: 53.]. A self-assembled film is an ordered supramolecular system formed by spontaneous adsorption of active molecules on heterogeneous interfaces through chemical bonds.

Developing more types of copolymer membranes and their simple preparation methods to meet specific application needs has become the direction of researchers' efforts.

Contents of the invention

The invention provides a method for synthesizing a copolymer film, a copolymer film prepared by the method and its application.

The present invention uses a catalyst to catalyze the polymerization and self-assembly of two types of film-forming monomers at two mutually immiscible liquid interfaces (liquid-liquid interface) to form an organic copolymer film. One type of film-forming monomers contains phenol. Another type of film-forming monomer is an organic compound containing at least two amino groups. The organic copolymer film can introduce other molecules and/or atoms to the surface of the film through secondary and/or multiple chemical and/or enzymatic reactions to obtain the expected chemical composition, molecular structure, physical properties and chemical properties, and achieve membrane performance to meet application requirements.

In order to improve the above technical problems, the present invention provides a method for synthesizing a copolymer film:

A film-forming monomer containing a phenolic hydroxyl group, a film-forming monomer containing at least two amino groups and a catalyst are dissolved in a water phase and/or a liquid phase that is immiscible with water, and polymerized at the interface of the two phases to form a film to obtain the result. The copolymer film.

Preferably, after the film-forming monomer containing phenolic hydroxyl group and the film-forming monomer containing at least two amino groups are dissolved in a solvent, a catalyst is added and mixed to obtain an aqueous phase, and the aqueous phase is contacted with a liquid phase that is immiscible with water. , polymerized to form a film at the interface of the two phases to obtain the copolymer film.

Preferably, the film-forming monomer containing phenolic hydroxyl groups is dissolved in a solvent, a catalyst is added and mixed to obtain an aqueous phase, the film-forming monomer containing at least two amino groups is dissolved in a liquid phase that is immiscible with water, and the aqueous phase is Enzymatic reaction occurs when the copolymer comes into contact with a liquid phase that is immiscible with water, and polymerizes to form a film at the interface of the two phases to obtain the copolymer film.

Preferably, the film-forming monomer containing at least two amino groups is dissolved in a solvent, a catalyst is added and mixed to obtain a water phase, the film-forming monomer containing phenolic hydroxyl groups is dissolved in a liquid phase that is immiscible with water, and the water phase is The copolymer film is brought into contact with a liquid phase that is immiscible with water, and is polymerized to form a film at the interface of the two phases to obtain the copolymer film.

Preferably, the catalyst is dissolved in the water phase, the film-forming monomer containing phenolic hydroxyl groups and the film-forming monomer containing at least two amino groups are dissolved in a liquid phase that is mutually immiscible with water, and the water phase and the film-forming monomer that are mutually incompatible with water are dissolved. When the miscible liquid phases contact, an enzymatic reaction occurs, and the copolymer film is polymerized to form a film at the interface of the two phases.

According to an embodiment of the present invention, the aqueous phase is an aqueous solution with a water content of not less than 50% in the total mass; film-forming monomers may be dissolved, and a certain amount of organic solvents soluble in water may be contained, and the organic solvent It can be methanol, ethanol, isopropyl alcohol, acetone, methyl formate, ethyl acetate, acetonitrile, tetrahydrofuran, N,N-dimethylformamide, 1,4-dioxane, dimethyl sulfoxide, dimethyl sulfoxide, etc. At least one of ethylene glycol butyl ether and diethylene glycol;

According to an embodiment of the present invention, the solvent is selected from water or a buffer solution, and the buffer solution and the buffer pair are pH buffers suitable to support the polymerization reaction and are not limited to any specific pH buffer. The buffer solution is preferably sodium acetate-acetic acid buffer solution, disodium hydrogen phosphate-citric acid buffer solution, potassium hydrogen phthalate-sodium hydroxide buffer solution, tartaric acid-sodium tartrate buffer solution, sodium citrate-citric acid buffer solution Solution, trisodium phosphate-phosphate buffer solution, sodium malonate-malonic acid buffer solution, sodium succinate-succinic acid buffer solution, phthalic acid-hydrochloric acid buffer solution, disodium hydrogen phosphate-sodium dihydrogen phosphate buffer solution , disodium hydrogen phosphate-potassium dihydrogen phosphate buffer solution, dipotassium hydrogen phosphate-sodium hydroxide buffer solution, Tris-hydrochloric acid buffer solution, boric acid-borax buffer solution, glycine-sodium hydroxide buffer solution.

According to an embodiment of the present invention, the liquid phase that is immiscible with water may be oil that is liquid at normal temperature, oil that is solid at a set temperature, or liquid metal. The oil that is liquid at room temperature may be at least one of vegetable oil, mineral oil and synthetic water-insoluble liquid. The vegetable oil may be peanut oil, soybean oil, flax oil, castor oil, rapeseed oil, corn oil, olive oil, flax oil, cinnamon oil, essential oil, etc. The mineral oil can be petroleum (crude oil), condensate, gasoline, kerosene, diesel, lubricating oil, transformer oil, engine oil, liquid paraffin, paraffin, and coal tar. The artificially synthesized water-insoluble liquids include various silicone oils, aromatic hydrocarbons (such as benzene, toluene, xylene, dichlorotoluene, bromobenzene, etc.), alkanes (such as pentadecane, tetradecane, tridecane, decacane, etc.). Dioxane, undecane, nonane, isooctane, hexane, chloroform, tetrachloromethane, carbon tetrachloride, carbon disulfide, etc.), cycloalkanes (such as cyclohexane, cyclopentane, cycloheptane, etc.) ), ethers (such as petroleum ether, butyl ether, etc.), esters (such as butyl oleate, butyl acetate, butyl palmitate, etc.), ketones (such as 2-nonanone, methyl isobutyl ketone, 3-hexanone etc.), at least one of organic acids (such as octanoic acid, etc.). The oil is solid at a set temperature, such as at least one of paraffin, cocoa butter, coconut oil, palm oil, and animal oil. The animal oil can be derived from pigs, cattle, sheep, horses, chickens, whales, insects (such as beeswax, insect wax, etc.), etc. The liquid metal, such as gallium, mercury, and low melting point alloys.

According to embodiments of the present invention, the catalyst may be at least one of a peroxidase and/or an oxidoreductase with laccase activity and/or an artificial enzyme with the aforementioned catalytic activity;

According to an embodiment of the present invention, the peroxidase may be selected from the group consisting of manganese peroxidase (MnP, EC 1.11.1.13), lignin peroxidase (LiP, EC 1.11.1.14) and at least one of chloroperoxidase (CPO, EC 1.11.1.10) and plant peroxidase (Plant peroxidase); the plant peroxidase can be horseradish peroxidase (Horseradish peroxidase) , HRP, EC 1.11.1.7), soybean peroxidase (SBP, EC 1.11.1.7), rice peroxidase (Rice peroxidase), cotton peroxidase (Cottonperoxidase), runner bean peroxidase Runner beans peroxidase, Garbanzo beans peroxidase, Guar beans peroxidase, Pea peroxidase, etc.

According to an embodiment of the present invention, the oxidoreductase with laccase activity is an enzyme classification EC 1.10 specified by the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology (IUBMB). 3.2 Included are any laccase (Laccase) from plants, animals, fungi, bacteria, yeast, or obtained through biotechnology, and/or any fragments derived from them that exhibit laccase activity, and/or exhibit similar activity Enzymes such as catecholoxidase (CO, EC 1.10.3.1) or any fragment derived from it that exhibits catechol oxidase activity, monophenol monooxygenase (EC 1.14.18.1) or At least one of any fragment from which it is derived that exhibits monophenol monooxidase activity, bilirubin oxidase (BOD, EC 1.3.3.5) or any fragment from which it is derived that exhibits bilirubin oxidase activity kind.

According to an embodiment of the present invention, the plant-derived laccase is selected from the group consisting of Anacardiaceae, Podocarpaceae, Aesculus sp., Acerpseudoplatanus, Catharanthus roseus Flower (Catharanthus roseus), carrot (Daucus carrota), Dichomitius squalens), ginkgo (Gingko biloba), apple (Malus pumila), Panasonic orchid (Monotropahypopithys), avocado (Persea americana), peach (Prunus persica), potato (Solanumtuberosum) ), rosemary (Rosmarinus officinalis), and Vinca minor extracts.

According to an embodiment of the invention, the laccase is preferably a fungal laccase. The fungal laccase is selected from the group consisting of Agaricus bisporus, Aspergillus nidulans, Botrytiscinerea, Ceriporiopsis subvermispora, Cerrena unicolor, Chaetomium thermophile, Cladosporium cladosporiodes, Coprinus cinereus, Coriolus hirsutus, Coriolusversicol, Fomes fomentarius ), Ganoderma lucidum, Glomerella cingulate, Heterobasidion annosum, Lactariuspiperatus, Lentinus tigrinus, Melanocarpus albomyces, Myceliophthora thermophila, Neurospora crassa, Panaeolus papilionaceus, Panaeolus sphinctrinus, Pleurotus ostreatus, Pleurotus spores (Podospora anserina), Polyporusbrumalis, Polyporus pinsitus, Polyporus versicolor, Pycnoporus cinnabarinus, Pyricularia orizae, Rhizoctonia praticola, Rhizoctonia solani ( Rhizoctonia solani), Rigidoporus lignosus, Russuladelica, Schizophyllum commune, Steccherinum ochraceum, Thelephora terrestris, Thielavia Laccases from Thielavia arenaria, Trameteshirsuta, Tramates versicolor and their variants.

Preferably, the fungal laccase is a laccase of Polyporus brumalis (the deposit number of the strain is CCTCC NO: M 2020809).

The laccase may also be an enzyme produced by a method comprising culturing a host cell transformed with a recombinant DNA vector carrying DNA encoding the function of said laccase in a culture medium under conditions that allow laccase expression sequence and then recover the laccase from the culture.

Compared with other enzymes, fungal laccase has the following advantages: (1) It can catalyze a wide range of substrates, and adding enzyme enhancers can catalyze more substrates; (2) Compared with bacterial laccase, it has a high redox potential ; (3) It is easier to realize industrial preparation; (4) The cost is lower; (5) No coenzyme is required; (6) The oxidant uses molecular oxygen, so there is no need to add hydrogen peroxide in the reaction; (7) Water is the only by-product The product will not form a useless enzyme-substrate complex; (8) It has glycosylation modification and the enzyme has good stability; (9) It is not highly sensitive to metal ions in the medium; (10) It is Many organic solvents have a certain tolerance.

According to an embodiment of the present invention, the structure of the compound containing phenolic hydroxyl groups is shown in Formula I:

Wherein, each R ₁ is the same or different and is independently selected from H, halogen, CN, NO ₂ , OH, SH, COOH, the following groups that are unsubstituted or substituted by one, two or more R _a1 : C _1-40 alkyl, C _2-40 alkenyl, C _2-40 alkynyl, C _3-40 cycloalkyl, C _3-40 cycloalkenyl, C _3-40 cycloalkynyl, C _6-20 aryl , 5-20 membered heteroaryl group, 3-20 membered heterocyclic group, -OR _1-2 , -SR _1-3 , -NR _1-4 R _1-5 , -C(O)R _1-6 , - OC(O)R _1-7 , -S(O) ₂ R _1-8 , -OS(O) ₂ R _1-9 , -P(O)R _1-10 R _1-11 , -N＝NR _{1 -12} ;

A ₁ exists or does not exist; when A ₁ exists, it is selected from the group consisting of unsubstituted or substituted by one, two or more R _b1 C _6-20 aryl groups, 5-20 membered heterocyclic groups connected to the benzene ring. Aryl, 5-20 membered heterocyclyl; or A ₁ is selected from chemical bonds, unsubstituted or optionally substituted by one, two or more R _c1 O, C(O), C(O)O, S , S(O) ₂ , N, C _1-6 alkylene, CH=N, N=N, CH=NN=CH, CH=CH-CO-CH ₂ -CO-CH=CH;

m is an integer from 0 to 5.

Each R _a1 , R _b1 , R _c1 is the same or different, and is independently selected from H, halogen, CN, OH, SH, oxo (=O), NO ₂ , COOH, -OR _1-2 , -SR _{1 -3} , -NR _1-4 R _1-5 , -C(O)R _1-6 , -OC(O)R _1-7 , -S(O) ₂ R _1-8 , -OS(O) ₂ R _1-9 , P(O)R _1-10 R _1-11 , C _{1-40 alkyl, C 2-40 alkyl ,} unsubstituted or optionally substituted by one, two or more R _1-12 Base, C _2-40 alkynyl group, C _3-40 cycloalkyl group, C _3-40 cycloalkenyl group, C _3-40 cycloalkynyl group, C _6-20 aryl group, 5-20 membered heteroaryl group, 3- 20-membered heterocyclyl;

Each of R _1-2 , R _1-3 , R _1-4 , R _1-5 , R _1-6 , R _1-7 , R _1-8 , R _1-9 , R _1-10 , R _{1- 11.} R _1-12 are the same or different, and are independently selected from H, halogen, CN, OH, SH, oxo (=O), NO ₂ , COOH, C _1-40 alkyl, C _2-40 alkenyl , C _2-40 alkynyl, C _3-40 cycloalkyl, C _3-40 cycloalkenyl, C _3-40 cycloalkynyl, C _6-20 aryl, 5-20 membered heteroaryl, 3-20 Heterocyclic group.

According to an embodiment of the present invention, Formula I has the structures shown in Formulas I-1 to I-13:

Wherein, R ₁ , m and R _c1 have the definitions mentioned above; B is selected from O or N substituted by R ₁ ; D, D ₁ , D ₂ and D ₃ are the same or different, and are independently selected from chemical bonds, O, _CH2 , C(O) or N-CO- _CH3 that is unsubstituted or optionally substituted by one, two or more _Rc1 ; E and _E1 are the same or different, independently selected from chemical bonds, O, C(O), C(O)O, S, S(O) ₂ , N, Se, P, C _1-6 substituents that are unsubstituted or optionally substituted with one, two or more R _c1 Alkyl group, CH=CH, CH=N, N=N, CH=NN=CH, CH=CH-CO-CH ₂ -CO-CH=CH; n is an integer from 0 to 5; F ₁ and F ₂ are the same Or different, independently selected from N, CH or O ⁺ ; R ₁ ', R ₁ ″ and R ₁ have the same definition.

Preferably, the compound of formula I-1 is selected from the group consisting of catechol (CAS: 120-80-9), 3-methylcatechol (CAS: 488-17-5), 3,4-dihydroxy Toluene (CAS:452-86-8), 4-ethylcatechol (CAS:1124-39-6), 4-tert-butyl-1,2-benzenediol (CAS:98-29-3 ), 3,5-di-tert-butyl-1,2-benzenediol (CAS:1020-31-1), 3-methoxycatechol (CAS:934-00-9), 3,4 -Dihydroxybenzaldehyde (CAS:139-85-5), 3,4-dihydroxystyrene (CAS:6053-02-7), 2',4'-dihydroxyacetophenone (CAS:89-84 -9), hydroxytyrosol (CAS:10597-60-1), dopamine (CAS:51-61-6) and its salts, 3-methacryloyldopamine (CAS:471915-89-6), epinephrine (CAS:51-43-4), norepinephrine and its salts, isoproterenol hydrochloride (CAS:51-30-9), methyl protocatechuate (CAS:2150-43-8) and other esters Class, ferrotitanium reagent (CAS:149-45-1), 3-bromo-5-tert-butyl-1,2-benzenediol, ethyl 3,4-dihydroxybenzoate (CAS:3943-89- 3) Esters such as ethyl 2,3-dihydroxybenzoate, 4-nitrocatechol (CAS: 3316-09-4), urushiol, Suillin (CAS: 103538-03-0 ), Grifolin (CAS: 6903-07-7), Ilicicolin B (CAS: 22581-07-3), Resorcinol (CAS: 108-46-3), 4-Chlororesorcinol (CAS: 95 -88-5), 2,6-dihydroxytoluene (CAS:608-25-3), 2,4-dihydroxybenzaldehyde (CAS:1995-1-2), 2',4'-dihydroxybenzene Ethyl ketone (CAS: 89-84-9), 2',6'-dihydroxyacetophenone (CAS: 699-83-2), 4-hexylresorcinol (CAS: 136-77-6), Cardol, terbutaline sulfate (CAS: 23031-32-5), hydroquinone (CAS: 123-31-9), tert-butylhydroquinone (CAS: 1948-33- 0), 2-chlorohydroquinone (CAS: 615-67-8), homogentisic acid (CAS: 451-13-8), 3,6-dihydroxyphthalonitrile (CAS: 4733-50 -0), 2,6-dimethoxyhydroquinone (CAS:15233-65-5), calcium dobesilate (CAS:20123-80-2), cannabidiol (CAS:25654-31 -3), Ocinaline sulfate (CAS:5874-97-5), cannabidiol (CAS:13956-29-1), phloroglucinol (CAS:6099-90-7), 2,4,6 -Esters such as trihydroxybenzaldehyde (CAS:487-70-7), ethyl 2,4,6-trihydroxybenzoate (CAS:90536-74-6), 6-hydroxydopamine hydrochloride (CAS:28094 -15-7), 6-hydroxydopamine hydrobromide (CAS: 636-00-0), 2,4,5-trihydroxyphenylalanine (CAS: 23358-64-7), pyrogallol ( CAS:87-66-1), 2',3',4'-trihydroxyacetophenone (CAS:528-21-2), 2,3,4-trihydroxybenzaldehyde (CAS:2144-08- 3), esters such as methyl gallate (CAS: 99-24-1), 4-bromophenol (CAS: 106-41-2), p-hydroxyphenylethylamine (CAS: 51-67-2), 4 -Methylphenol (CAS:106-44-5), 4-chlorophenol (CAS:106-48-9), 4-methoxyphenol (CAS:150-76-5), 2,3-dimethyl Oxyphenol (CAS:5150-42-5), 4-ethylphenol (CAS:123-07-9), vanillin (CAS:121-33-5), isovanillin (CAS:621-59- 0), vanillyl alcohol (CAS: 498-00-0), guaiacol (CAS: 90-05-1), o-acetaminophen (CAS: 614-80-2), acetaminophen (CAS: 103-90-2), 3,4,5-trimethoxyphenol (CAS:642-71-7), 4-(2'-thiazolyl azo)resorcinol (CAS:2246-46-0 ), esters such as ethyl 2,3,4-trihydroxybenzoate, benserazide hydrochloride (CAS: 14919-77-8), etc.

Preferably, the compound of I-2 is selected from 3',4-dihydroxyflavone (CAS:4143-64-0), 6,7-dihydroxyflavone (CAS:38183-04-9), 7,8 -Dihydroxyflavone (CAS:38183-03-8), acacetin (CAS:480-44-4), coine (CAS:480-40-0), kaempferol (CAS:491-54-3) , lycopene (CAS: 520-12-7), isorhamnetin (CAS: 480-19-3), myerin (CAS: 520-34-3), syringantin (CAS: 4423-37- 4), Rhododendron (CAS: 491-71-4), Rubin (CAS: 480-15-9), Rhododendron (CAS: 529-51-1), Tamarixanthin (CAS: 603 -61-2), 5,7-dihydroxy-2-(4-hydroxyphenyl)-6,8-dimethoxy-4H-1-benzopyran-4-one (CAS: 4323-80 -2), fisetin (CAS: 528-48-3), scaprosin (CAS: 18398-74-8), apigenin (CAS: 520-36-5), galangalin (CAS: 548 -83-4), begaine (CAS:491-67-8), quercetin dihydrate (CAS:6151-25-3), 6-methoxylutein (CAS:520-11- 6), 3-O-methylquercetin (CAS: 1486-70-0), morin (CAS: 480-16-0), luteolin (CAS: 491-70-3), quercetin (CAS: 117-39-5), myricetin (CAS: 529-44-2), basil (CAS: 437-64-9), 5,7-dihydroxy-6,8,4'-trimethyl Oxyflavonoids (CAS: 10176-66-6), hemanoxin (CAS: 56003-01-1), kaempferol (CAS: 520-18-3), scutellarin (CAS: 529-53-3 ), 7,3',4'-trihydroxy-3,8-dimethoxyflavone, 3,7,3',4'-tetrahydroxy-8-methoxyflavone, 3,7,8,3 ',4'-pentahydroxyflavone, etc.

Preferably, the compound of I-3 is selected from the group consisting of texaxanthin (CAS: 897-46-1), daidzin (CAS: 486-66-8), and corystine A (CAS: 491-80 -5), 4',6,7-trihydroxyisoflavone (CAS:17817-31-1), genistein (CAS:446-72-0), oropol (CAS:480-23-9 ), cardine (CAS: 20575-57-9), pulin (CAS: 552-59-0), etc.

Preferably, the compound of I-4 is selected from esculetin (CAS: 305-01-1), 4-methyl esculetin (CAS: 529-84-0), etc.

Preferably, the compound of I-5 is selected from the group consisting of hesperetin (CAS: 520-33-2), naringenin (CAS: 67604-48-2), xanthophyllin (CAS: 20725-03-5), Citrinin (CAS: 480-41-1), epiafcatechin (CAS: 24808-04-6), acaitin (CAS: 4382-33-6), dihydroquercetin (CAS: 480 -18-2), dihydromyricetin (CAS:27200-12-0), eriodictyol (CAS:552-58-9), fisetinol (CAS:490-49-3), (+ )-Rhobidanol (CAS: 17445-90-8), gallocatechin (CAS: 3371-27-5), 4',7-dihydroxyisoflavane (CAS: 531-95-3), afurol Tea essence (CAS: 2545-00-8), (+)-catechin (CAS: 154-23-4), epigallocatechin (CAS: 970-74-1), epicatechin, phoenix Descendol, Fisetinol, protogeranidine monomer, protofesserdine monomer, protopterinol monomer, leucocyaninedine monomer, protocyanidine monomer, delphinium monomer, nasturtium Catechin, mesquitin, acetate, 7,8,4'-trihydroxyflavanol, 3,4-dihydroxyflavanol, etc.

Preferably, the compound of I-6 is selected from the group consisting of anthocyanins (CAS: 528-58-5), delphinidin chloride (CAS: 528-53-0), and pelargonidin (CAS: 134-04-3 ), cinnamon acetate chloride (CAS: 23130-31-6), petunia pigment (CAS: 1429-30-7), malvacin (CAS: 643-84-5), etc.

Preferably, the compound of I-7 is selected from p-phenylphenol (CAS: 92-69-3), azo violet (CAS: 74-39-5), 2,4-dihydroxybenzophenone ( CAS:131-56-6), phenethylresorcinol (CAS:85-27-8), xanthohumol (CAS:6754-58-1), resveratrol (CAS:501-36- 0), rosmarinic acid (CAS: 20283-92-5), phloretin (CAS: 60-82-2), nordihydroguaiaretic acid (CAS: 500-38-9), tetrahydroxy Stilbene, piceatannol (CAS:10083-24-6), curcumin (CAS:458-37-7), bisphenol A (CAS:80-05-7), 2',2'-dihydroxyconjugate Benzene (CAS: 1806-29-7), 4,4'-dihydroxybiphenyl (CAS: 92-88-6), phenolphthaloline (CAS: 81-90-3), thiobisdichlorophenol (CAS: 97-18-7), dichlorophenol (CAS: 97-23-4), 2,2'-dihydroxybenzophenone (CAS: 835-11-0), 4,4'-dihydroxydiphenyl Methyl ketone (CAS: 611-99-4), 4-nitrobenzene azo (CAS: 607-96-5), 2,2'-dihydroxy-4-methoxybenzophenone (CAS: 131 -53-3), honokiol (CAS: 35354-74-6), magnolol (CAS: 528-43-8), diethylstilbestrol (CAS: 5635-50-7), diethylstilbestrol (CAS: 6898-97-1), 3,3',5,5'-(tetraisopropyl)biphenyl 4,4'-diphenol (CAS: 2416-95-7), 2,2'-dihydroxybiphenyl Phenyl ether (CAS: 15764-52-0), dobutamine hydrochloride (CAS: 49745-95-1), witch hazel tannin (CAS: 469-32-9), 2,,3,3' ,4,4',5-hexahydroxydiphenylsulfone, 3,3',4,4',5,5'-hexahydroxydiphenylsulfone, 2,2',3,3',4,4'- Hexahydroxydiphenylsulfone, hexahydroxybiphenyldioic acid, etc.

Preferably, the compound of I-8 is selected from the group consisting of 2,3-dihydroxynaphthalene (CAS: 92-44-4), 1,2-dihydroxynaphthalene (CAS: 574-00-5), 1,3-dihydroxynaphthalene (CAS: 574-00-5), Dihydroxynaphthalene (CAS: 132-86-5), 1,5-dihydroxynaphthalene (CAS: 83-56-7), 1,6-dihydroxynaphthalene (CAS: 575-44-0), 1,7 -Dihydroxynaphthalene (CAS:575-38-2), 1,4-dihydroxynaphthalene (CAS:571-60-8), 2,7-dihydroxynaphthalene (CAS:582-17-2), 6, Sodium 7-dihydroxy-2-naphthalene sulfonate (CAS: 135-53-5), 3,5-dihydroxy-2-naphthoic acid (CAS: 89-35-0), calcium carboxylic acid (CAS: 3737- 95-9), calcium magnesium reagent (CAS: 3147-14-6), eletrochrome blue black R (CAS: 2538-85-4), chromium 2R (CAS: 4197-07-3), dicarboxylic acid Naphthoic acid (CAS: 130-85-8), beryllium reagent II (CAS: 51550-25-5), chromotropic acid (CAS: 148-25-4), chromotropic acid disodium salt (CAS: 5808-22-0 ), calcium pigment (CAS: 3810-39-7), eletrochrom blue black B (CAS: 3564-14-5), eletrochrom blue SE (CAS: 1058-92-0), etc.

Preferably, the compound of I-9 is selected from shikonin (CAS:517-89-5) and the like.

Preferably, the compound of I-10 is selected from the group consisting of aloe purpurin (CAS: 481-72-1), anthranol (CAS: 1143-38-0), 1,5-dihydroxyanthraquinone (CAS: 117 -12-4), 1,4-dihydroxyanthraquinone (CAS:81-64-1), 1,8-dihydroxyanthraquinone (CAS:117-10-2), 10-acetyl-3,7 -Dihydroxyphenazine (CAS:119171-73-2), anthracinol (CAS:117-12-4), chrysophanol (CAS:481-74-3), rhein (CAS:478-43-3 ), 1,2,3-trihydroxyanthraquinone (CAS:602-64-2), purpurin (CAS:81-54-9), alizarin red (CAS:130-22-3), alizarin Complex indicator (CAS:3952-78-1), alizarin (CAS:72-48-0), alizarin red S (CAS:130-22-3), mitoxantrone hydrochloride (CAS:70476- 82-3), fluorescein (CAS: 518-44-5), emodin (CAS: 518-82-1), lac chromic acid (CAS: 60687-93-6), quinone alizarin (CAS: 81 -61-8), α-mangostin (CAS: 6147-11-1), etc.

Preferably, the compound of I-11 is selected from gallic blue (CAS: 1562-85-2), celestite blue (CAS: 1562-90-9), etc.

Preferably, the compound of I-12 is selected from 1,8,9-trihydroxyanthracene (CAS: 480-22-8) and the like.

Preferably, the compound of I-13 is selected from p-hydroxybenzoic acid (CAS: 99-96-7), m-hydroxybenzoic acid (CAS: 99-06-9), vanillic acid (CAS: 121-34-6 ), isovillic acid (CAS: 645-08-9), 4-hydroxy-2-methoxybenzoic acid (CAS: 90111-34-5), 3-hydroxy-5-methoxybenzoic acid (CAS: 19520 -75-3), salicylic acid (CAS:69-72-7), p-aminosalicylic acid (CAS:65-49-6), m-aminosalicylic acid (CAS:89-57-6), 3 -Methyl salicylic acid (CAS: 83-40-9), 5-methyl salicylic acid (CAS: 89-56-5), 3-methoxysalicylic acid (CAS: 877-22-5) , 4-methoxysalicylic acid (CAS:2237-36-7), 5-methoxysalicylic acid (CAS:2612-02-4), 6-methoxysalicylic acid (CAS:3147- 64-6), 3,5-diiodosalicylic acid (CAS: 133-91-5), syringic acid (CAS: 530-57-4), paeonol (CAS: 552-41-0), cumin Such as acid (CAS: 126633-45-2), ginkgolic acid (CAS: 22910-60-7), diflunisal (CAS: 22494-42-4), gentisic acid (CAS: 490-79-9) , Pyrocatechuic acid (CAS: 303-38-8), resorcinol acid (CAS: 89-86-1), 3,5-dihydroxybenzoic acid (CAS: 99-10-5), 2,6- Dihydroxybenzoic acid (CAS:303-07-1), gallic acid (CAS:149-91-7), ferulic acid (CAS:1135-24-6), p-coumaric acid (CAS:501-98- 4), tyrosine, sinapinic acid (CAS:530-59-6), 4-hydroxymandelic acid (CAS:1198-84-1), 2-hydroxyphenylglycine (CAS:25178-38-5), coffee acid (CAS: 331-39-5), bisphenolic acid (CAS: 126-00-1), protocatechuic acid (CAS: 99-50-3), danshensu (CAS: 76822-21-4), Dopa, methyldopa (CAS:555-30-6), 2,4,6-trihydroxybenzoic acid (CAS:83-30-7), etc.

According to an embodiment of the present invention, Formula I also includes substances in which there are two or more structural units represented by Formulas I-1 to I-13, such as golden pine biflavonoids (CAS: 521-34-6), Cinnamon tannin (CAS: 86631-38-1), 2,2'-dihydroxy-1,1'-binaphthyl (CAS: 602-09-5), 6,6'-dithiodinaphthol ( CAS: 6088-51-3), hypericin (CAS: 548-04-9), pyrantel pamoate (CAS: 22204-24-6), 2,3-dihydropyrobioflavone (CAS :34421-19-7), taikobioflavone (CAS: 1617-53-4), isoginkgobiflavonoids (CAS: 548-19-6), desmethyl ginkgobiflavonoids (CAS: 521-32-4 ), 5'-methoxyginkgobislavone (CAS:77053-35-1), ginkgo biloba (CAS:481-46-9), proanthocyanidin B1 (CAS:20315-25-7), proanthocyanidin B2 (CAS: 29106-49-8), proanthocyanidin B3 (CAS:23567-23-9), proanthocyanidin B4 (CAS:29106-51-2), proanthocyanidin B5 (CAS:12798-57-1), proanthocyanidin B6 (CAS:12798- 58-2), procyanidin B7 (CAS: 12798-59-3), proanthocyanidin B8 (CAS: 12798-60-6), (-)-epigallocatechin gallate (CAS: 989-51-5) , epicatechin gallate (CAS: 1257-08-5), catechin gallate (CAS: 130405-40-2), epigallocatechin gallate (CAS: 4233-96-9) , Protodelphinidin B2-3'-O-gallic acid, dehydrocatechin B, tetrabromotannins, procyanidine B-2,3'-O-gallate, procyanidine B-2,3,3'-di-O-gallate, protodelphinidin B1 (CAS: 78362-04-6), protodelphinidin B2 (CAS: 87392-61-8), protodelphinidin Delphinidin B3 (CAS:78362-05-7), protodelphinidin B4 (CAS:68964-95-4), etc.

According to an embodiment of the present invention, the compounds of formula I also include: saxifragetin (CAS: 477-90-7), wogonin (CAS: 632-85-9), irisin (CAS: 548- 77-6), licorice flavonoids (CAS: 59870-68-7), ellagic acid (CAS: 476-66-4), catalexin (CAS: 82-08-6), Caesin J (CAS: 99217 -67-1), oxidized hematoxylin (CAS: 475-25-2), pyrogallol (CAS: 569-77-7), Gnetin A (CAS: 82084-87-5), pyrogallic acid red (CAS:32638-88-3), hematoxylin (CAS:517-28-2), catechol violet (CAS:115-41-3), 1-naphtholphthalein (CAS:596-01-0 ), Arsenazo I (CAS:520-10-5), Arsenazo III (CAS:1668-00-4), Phenolphthalein (CAS:77-09-8), Phenol Red (CAS:143-74- 8), Tetraiodophenol sulfonphthalein (CAS: 4430-24-4), safflower yellow (CAS: 36338-96-2), fluorescein (CAS: 2321-07-5), xylenol blue (CAS :125-31-5), 4',5'-dibromofluorescein (CAS:596-03-2), rose acid (CAS:633-00-1), thymol blue (CAS:76- 61-9), Alizarin Violet (CAS: 2103-64-2), Glycyrrhizin (CAS: 60008-03-9), Cyperin, Cyanine, Delphinium, HeimiolA, Balanicarpolα-H, Hydrodicatechin A, fulvic acid (CAS: 479-66-3) and its salts, mycophenolic acid (CAS: 24280-93-1), alizarin blue S (CAS: 66675-89-6), Apomorphine hydrochloride (CAS:41372-20-7), (4E)-6-(4,6-dihydroxy-7-methyl-3-oxo-1,3-dihydroisobenzofuran-5 -yl)-4-methyl-4-hexenoic acid 2-(morpholin-4-yl)ethyl ester (CAS: 1322681-36-6), (+)-Boldine base (CAS: 476-70 -0), pseudomorphine (CAS:125-24-6), myostatin (CAS:57-94-3), silibinin (CAS:22888-70-6), isoniazid Foamycin hydrazone (CAS: 13292-53-0), daunorubicin (CAS: 20830-81-3), rifampicin (CAS: 13292-46-1), 3-formylrifamycin SV (CAS: 13292-22-3), rifamycin SV, rifaximin (CAS: 80621-81-4), doxorubicin hydrochloride (CAS: 25316-40-9), epirubicin hydrochloride (CAS:56390-09-1), Rifapentine (CAS:61379-65-5), Doxorubicin (CAS:54193-28-1), Rifamide (CAS:2750-76-7) , vancomycin and its salts, norvancomycin hydrochloride (CAS: 213997-73-0), flavan-3-en-3-ol, teicoplanin (CAS: 61036-62-2), Malibatol A , Malibatol B (CAS:204644-72-4), Suffruticosol A, Suffruticosol B, Vaticanol C, etc.

According to an embodiment of the invention, the compounds of formula I also include: polyphenols.

According to an embodiment of the present invention, the polyphenols include: proanthocyanidin (CAS: 4852-22-6), proanthocyanidin A1 (CAS: 103883-03-0), proanthocyanidin A2 (CAS: 41743-41-3), proanthocyanidin A4 (CAS :111466-29-6), procyanidin C1 (CAS: 37064-30-5), proanthocyanidin C2 (CAS: 37064-31-6), cinnamon tannin B1 (CAS: 88082-60-4), cinnamon tannin B2 (CAS:88038-12-4), theaflavins (CAS:4670-05-7), tea polyphenols (CAS:84650-60-2), tannic acid (CAS:1401-55-4), wood Ephedra tannins (CAS: 79786-01-9), ephedra tannins (CAS: 81739-27-7), Davuriciin T1 (CAS: 137371-86-9), pomegranate peel tannins (CAS: 65995-64- 4), Gemin A (CAS: 137371-86-9), Diyumin H2 (CAS: 82200-04-2), Artemisol A (CAS: 124027-58-3), Hopeaphenol (CAS: 17912-85 -5), geranin (CAS: 60976-49-0), oak ellagitin (CAS: 36001-47-5), chestnut ellagitin (CAS: 24312-00-3), Corile Acid (CAS: 18942-26-2), Casuarina tannin (CAS: 79786-00-8), α-glucotin (CAS: 62218-13-7), ε-glucotin (CAS: 62218-08- 0), δ-glucotin, Gnetin C(CAS:84870-54-2), Gnetin D(CAS:84870-53-1), Gnetin J(CAS:152511-23-4), Pallidol(CAS:105037- 88-5), miyamotool C (CAS:109605-83-6), salvianolic acid B (CAS:115939-25-8), humic acid (CAS:1415-93-6) and its salts, nitro Humic acid (Nitro humic acid), xanthophyllol, dehydrogenated ellagic acid, acetate acid and its esters, biflavanocyanin, oolonganocyanin, pomegranate, polyprotodelphinidine, etc.

According to embodiments of the present invention, compounds of formula I also include: compounds formed by combining compounds of formulas I-1 to I-13 with quinic acid and/or sugar groups and/or sugar acids, such as arbutin (CAS: 497-76-7), scutellarin (CAS: 27740-01-8), rhein (CAS: 155-58-8), chlorogenic acid (CAS: 327-97-9), narcissin (CAS :604-80-8), Puerarin (CAS:3681-99-0), Hesperidin (CAS:520-26-3), Neohesperidin (CAS:13241-33-3), Dior Siming (CAS: 520-27-4), aloin (CAS: 5133-19-7), forsythiaside E (CAS: 93675-88-8), angloside C (CAS: 115909-22 -3), kaempferol-7-O-D-glucoside (CAS: 16290-07-6), naringin (CAS: 10236-47-2), apigenin-7-glucoside (CAS: 578-74- 5), Paeoniflorin-3,5-diglucoside (CAS: 132-37-6), vetch glycoside (CAS: 480-10-4), alfudoside (CAS: 482-39-3), Kaempferol-3-O-rutinoside (CAS: 17650-84-9), Kaempferol-3-O-glucose (1-2) rhamnoside (CAS: 142451-65-8), Diocyanin (CAS :55804-74-5), isorhamnetin-3-O-D-glucoside (CAS:5041-82-7), syringantin-3-O-D-rutinoside (CAS:53430-50-5), mignonette Glucoside-4'-glucoside (CAS: 6920-38-3), astilbin (CAS: 29838-67-3), glucoside chloride paeoniflorin (CAS: 6906-39-4), malvacin- 3-O-galactoside (CAS: 30113-37-2), malvidin-3-O-arabinoside (CAS: 28500-04-1), phlorizin (CAS: 60-81-1), Neohesperidin dihydrochalcone (CAS:20702-77-6), rutin (CAS:153-18-4), trimasin II (CAS:81571-72-4), catechin- 7,3'-O-β-D glucose, Ginkgetin-7-O-D-glucopyranoside, Isoginkgetin-7-O-D-glucopyranoside 7-O-D-glucopyranoside), quercetin-4-methoxy-3'-D-glucoside, catechin-4'-O-β-D glucose, trimasol II, agrimonin, Stilacin phenazine, tea flavonoids, astaxanthin, masanin, polyflavanol oxyglycoside, polyflavanol carbon glycoside, quercetin-3-O-β-D-glucose, 3-O- {2-O-[6-O-(рD-glucosyl)-O-trans-coumaroyl]-D-glucosyl}-(L-rhamnosyl)quercetin, rhodophyllin A, rhodonin B, rhodin C, rhodin D, rhodin E, rhodin F, rhodin G, lutein 7-O-rhamnoglucoside, lutein 7-O-glucoside, genistein Flavonoid 7-O-rhamnose glucoside, xanthophyllin-3-O-glucoside, oropol 7-O-rhamnose glucoside, 3-O-[2-O,6-O bis(L -Rhamnosyl)-D-glucosyl]isorhamnetin, quercetin-4'-O-β-D-glucose-6”-gallate, quercetin-3-O-α-arabinate Sugar-2”-gallate, 2,3-O hexahydroxydiphenylglucose, etc. The sugar base includes glucose, xylose, galactose, arabinose, rhamnose, mannose, glucuronic acid, rutose, gentiobiose, sophorose, neohesperiose, acaciabiose, lactose, sophora Trisaccharide, caffeoylglucose, 2-acetylglucose, and combinations thereof.

Preferably, the film-forming monomer containing phenolic hydroxyl groups is selected from the group consisting of 1,3-dihydroxynaphthalene, p-hydroxybenzoic acid, ferulic acid, 1,6-dihydroxynaphthalene, shikonin, and 3-methylsalicylic acid. Acid, p-coumaric acid, 4,4'-dihydroxybiphenyl, bisphenol A, chlorogenic acid, 3-methoxysalicylic acid, gallic acid, 4',6,7-trihydroxyisoflavone, aroma Leaf lignin, tannic acid, 3,5-diiodosalicylic acid, gentisic acid, resveratrol, aescin, p-aminosalicylic acid, caffeic acid, ellagic acid, golden pine biflavonoids, anthocyanins , syringic acid, tyrosine, guaiacol, 2,6-dihydroxytoluene, catechol, hydroxytyrosol, hesperetin, 2,4,6-trihydroxybenzaldehyde, coine, iris yellow At least one kind selected from the group consisting of cinnamon, narcissin, rhein, galloblue, 1,8,9-trihydroxyanthracene, urushiol, and tert-butylhydroquinone.

According to an embodiment of the present invention, the film-forming monomer containing at least two amino groups has a structure represented by Formula II:

Each R ₂ is the same or different and is independently selected from the group consisting of H, halogen, CN, NO ₂ , NO, OH, SH, COOH, unsubstituted or substituted with one, two or more R _a2 : C _1-40 alkyl, C _2-40 alkenyl, C _2-40 alkynyl, C _3-40 cycloalkyl, C _3-40 cycloalkenyl, C _3-40 cycloalkynyl, C _6-20 aryl , 5-20-membered heteroaryl group, 3-20-membered heterocyclic group, -OR _2-2 , -SR _2-3 , -NR _2-4 R _2-5 , -C(O)R _2-6 , - OC(O)R _2-7 , -S(O) ₂ R _2-8 , -OS(O) ₂ R _2-9 , P(O)R _2-10 R _2-11 ;

A ₂ is selected from C _1-40 alkyl, C _2-40 alkenyl, C _2-40 alkynyl, C ₃ which is unsubstituted or substituted by one, two or more R _b2 and connected to the benzene ring. _-40 cycloalkyl, C _3-40 cycloalkenyl, C _3-40 cycloalkynyl, C _6-20 aryl, 5-20-membered heteroaryl, 3-20-membered heterocyclyl; or A ₂ is selected from Chemical bond, O, C(O), C(O)O, S, S(O) ₂ , N, C _1-6 alkylene group that is unsubstituted or optionally substituted by one, two or more R _c2 , CH=N, N=N, CH=NN=CH, CH=CH-CO-CH ₂ -CO-CH=CH;

p is an integer from 0 to 12;

Each R _a2 , R _b2 , R _c2 is the same or different, and is independently selected from H, halogen, CN, OH, SH, oxo (=O), =NH, NO ₂ , COOH, C _1-40 alkyl , C _2-40 alkenyl, C _2-40 alkynyl, C _3-40 cycloalkyl, C _3-40 cycloalkenyl, C _3-40 cycloalkynyl, C _6-20 aryl, 5 to 20 yuan Heteroaryl group, 3-20 membered heterocyclic group, -OR _2-2 , -SR _2-3 , -NR _2-4 R _2-5 , -C(O)R _2-6 , -OC(O)R _2-7 , -S(O) ₂ R _2-8 , -OS(O) ₂ R _2-9 , P(O)R _2-10 R _2-11 ;

Each of R _2-2 , R _2-3 , R _2-4 , R _2-5 , R _2-6 , R _2-7 , R _2-8 , R _2-9 , R _2-10 , R _{2- 11} are the same or different, independently selected from H, halogen, NH ₂ , CN, OH, SH, oxo (=O), NO ₂ , COOH, C _1-40 alkyl, C _2-40 alkenyl, C _2-40 alkynyl, C _3-40 cycloalkyl, C _3-40 cycloalkenyl, C _3-40 cycloalkynyl, C _6-20 aryl, 5-20-membered heteroaryl, 3-20-membered heteroaryl ring base.

According to an embodiment of the present invention, Formula II has the structures shown in Formulas II-1 to II-10:

Among them, R ₂ , p and R _c2 have the definitions mentioned above; q is an integer from 0 to 12; R ₂ ', R ₂ ″ have the same definitions as R 2 ; and R ₂ , _{R 2 '} , R ₂ ″ contains at least two amino groups;

Each D ₂ , D ₃ , and D ₄ are the same or different, and are independently selected from N, unsubstituted or CH substituted by R ₂ , and N ⁺ ;

Each E ₂ , E ₃ , E ₄ is the same or different, and is independently selected from O, C(O), C(S) that are chemically bonded, unsubstituted or optionally substituted by one, two or more R _c2 , C(O)O, S, S(O) ₂ , N, C _1-6 alkylene, CH=N, N=N, CH=NN=CH, C _0-6 alkylene/alkenyl- CO-C _1-6 alkylene-CO-C _0-6 alkylene/alkenyl, C _0-6 alkylene/alkenyl-CO-NH-C _0-6 alkylene-CO-C _{0 -6} alkylene/alkenyl, C _0-6 alkylene/alkenyl -NH-C _0-6 alkylene -NH-C _0-6 alkylene/alkenyl, C _0-6 alkylene /Alkenyl-C(O)OC _0-6 alkylene-C(O)OC _0-6 alkylene/alkenyl.

Each F ₂ is the same or different and is independently selected from O, S.

Preferably, the compound of II-1 is selected from p-phenylenediamine (CAS: 106-50-3) and its salts, p-toluenediamine (CAS: 95-70-5), 2,3-dimethyl -P-phenylenediamine (2,3-Dimethyl-1,4-benzenediamine), 2,6-Dimethyl-p-phenylenediamine (2,6-Dimethyl-1,4-benzenediamine), 2,3-diamine Ethyl-p-phenylenediamine (2,3-Diethyl-1,4-benzenediamine), 2,5-dimethyl-p-phenylenediamine (CAS: 6393-01-7), 2-hydroxyethyl-p-phenylenediamine Diamine sulfate (CAS:93841-25-9), 2-Fluoro-1,4-benzenediamine, 2-Isopropyl-1,4-benzenediamine 4-benzenediamine), 2-Hydroxymethyl-1,4-benzenediamine, 2-Hydroxyethoxy-1,4-benzenediamine, 2 -Acetylaminoethoxy-1,4-benzenediamine (2-Acetylaminoethoxy-1,4-benzenediamine), N-(4-aminobenzyl)biguanide, 2-nitro-1,4-phenylenediamine (CAS:5307 -14-2), 2-chloro-1,4-phenylenediamine (CAS: 615-66-7), 2,5-dichloro-1,4-phenylenediamine (CAS: 20103-09-7) , 2,5-toluenediamine sulfate (CAS: 615-50-9), o-phenylenediamine (CAS: 95-54-5) and its salts, 4-chloro-1,2-phenylenediamine (CAS: 95-83-0), Amiloride hydrochloride (CAS:2016-88-8), 4,5-dichloro-1,2-phenylenediamine (CAS:5348-42-5), 3,4- Diaminotoluene (CAS: 496-72-0), 3-nitro-1,2-phenylenediamine (CAS: 3694-52-8), 4-nitro-1,2-phenylenediamine (CAS: 99-56-9), 3,4-diaminobenzoic acid (CAS:619-05-6), m-phenylenediamine (CAS:108-45-2) and its salts, 4-chloro-1,3- Phenylenediamine (CAS:5131-60-2), 2,6-diaminotoluene (CAS:823-40-5), 2,4-diaminoanisole (CAS:615-05-4) and its Salts, 3,5-diaminobenzoic acid (CAS:535-87-5) and its salts, 2,4-diaminophen dihydrochloride (CAS:137-09-7), 2,4-diaminobenzenesulfonate Acid (CAS:88-63-1), 2,3-diaminopyridine (CAS:452-58-4), 2,4,6-triaminopyrimidine (CAS:1004-38-2), 2,4 -Diamino-6-methyl-1,3,5-triazine (CAS:542-02-9), melamine (CAS:108-78-1), 6-phenyl-1,3,5-triazine Azine-2,4-diamine (CAS:91-76-9), diaminaveratridine (CAS:5355-16-8), 2,4,5-triamino-6-hydroxypyrimidine sulfate (CAS :35011-47-3), trimethoprim (CAS:738-70-5), phenazopyridine hydrochloride (CAS:136-40-3), pyrimethamine (CAS:58-14-0) , isoladine maleate (CAS:84504-69-8), trimethoprim (CAS:738-70-5), 4-amino-6-chlorobenzene-1,3-disulfonamide (CAS :121-30-2), m-aminophenylsemicarbazide, etc.

Preferably, the compound of II-2 is selected from the group consisting of 3,4-diaminobenzophenone (CAS: 39070-63-8), basic orange 2 (CAS: 532-82-1), o-toluidine (CAS:119-93-7) and its salts, 3,3'-diaminobenzidine (CAS:91-95-2) and its salts, 4,4'-diaminobenzidine (CAS:92-87 -5) and its salts, 3,3',5,5'-tetramethylbenzidine dihydrochloride (CAS:54827-17-7), bis(4-amino-3-chlorophenyl)methane (CAS: 101-14-4), 4,4'-diaminodiphenylmethane (CAS:101-77-9), 4,4'-diaminodiphenyl ether (CAS:101-80-4), 4,4 '-Diaminodiphenyl sulfide (CAS: 139-65-1), 4,4'-diaminobenzophenone (CAS: 611-98-3), 4,4'-diaminobibenzyl (CAS :621-95-4), 4,4'-diaminodiphenyl sulfone (CAS: 80-08-0), 4,4'-diaminodiphenylamine and its salts, 3,3-dichlorobenzidine ( CAS:91-94-1), 3,3`-dimethoxybenzidine (CAS:119-90-4), 3,3`-dimethyl-4,4`-diaminodiphenylmethane ( CAS:838-88-0), 4,5' diaminodiphenyl sulfide, etc.

Preferably, the II-3 compound is selected from ethylenediamine (CAS: 107-15-3) and its salts, 1,2-propanediamine (CAS: 78-90-0), 1,3-propanediamine Diamine (CAS: 109-76-2), 1,4-diaminobutane (CAS: 110-60-1), 1,5-pentanediamine (CAS: 462-94-2), 1,6 -Hexamethylenediamine (CAS:124-09-4), 1,7-heptanediamine (CAS:646-19-5), 1,8-octanediamine (CAS:373-44-4), 1, 10-Decanediamine (CAS: 646-25-3), diethylenetriamine (CAS: 111-40-0), triethylenetetramine (CAS: 112-24-3), tetraethylenepentamine (CAS: 112-57-2), pentaethylene hexaamine (CAS: 4067-16-7), hexaethylene heptaamine, spermamidine (CAS: 124-20-9), high precision amidine (CAS: 4427-76-3) , Aminopropylcadaverine (CAS:56-19-9), Norspermine (CAS:56-18-8), Spermine (CAS:71-44-3), Thermophilic Spermine (CAS:70862 -11-2), thermophilic pentylamine (CAS: 13274-42-5), thermophilic pentylamine (CAS: 84807-66-9), thermophilic hexylamine (CAS: 63833-74-9), thermophilic Thermal hexylamine (CAS: 133416-04-3), 3,3'-iminodipropylamine (CAS: 56-18-8), N,N'-bis(3-aminopropyl)-1,3 -Propylenediamine (CAS:4605-14-5), urea (CAS:57-13-6), thiophenylurea, biuret (CAS:108-19-0), thiosemicarbazide (CAS:79- 19-6), acrylamide methylene urea, guanidine (CAS:90332-86-8), biguanide nitrate (CAS:22817-07-8), glycylglutamine (CAS:13115-71-4) , Alanyl glutamine (CAS: 39537-23-0), glutamine, N-(2-guanidinethyl)guanidine (CAS: 44956-51-6), 2,6-diaminopimelic acid (CAS:583-93-7), 2,4-diaminobutyric acid dihydrochloride, tranquillizer (CAS:57-53-4), 2,3-diaminopropionic acid (CAS:515-94-6 ), Aminoguanidinesulfonate, Oxalamide, Triaminoguanidinehydrochloride, Aminoguanidine, agmatine (CAS: 306-60-5), etc.

Preferably, the II-4 compound is selected from proflavin (CAS: 92-62-6) and its salts, safranin T (CAS: 477-73-6), etc.

Preferably, the compound of II-5 is selected from 6-hydroxy-2,4,5-triaminopyrimidine (CAS:1004-75-7), 2,4-diamino-6-hydroxypyrimidine (CAS:1956 -6-4) etc.

Preferably, the compound of II-6 is selected from 5,6-diamino-1,3-dimethyluracil (CAS:5440-00-6), 4,5-diamino-6-hydroxy-2 -Mercaptopyrimidine (CAS:1004-76-8), etc.

Preferably, the compound of II-7 is selected from the group consisting of 2,3-diaminonaphthalene (CAS:771-97-1), 1,8-diaminonaphthalene (CAS:479-27-6), 1,5- Diaminonaphthalene (CAS:2243-62-1), triamterene (CAS:396-01-0), methotrexate (CAS:59-05-2), dihydralazine sulfate (CAS:7327 -87-9) etc.

Preferably, the II-8 compound is selected from 1,2-diaminocyclohexane (CAS: 694-83-7), strepamine (CAS: 488-52-8), etc.

Preferably, the compound of II-9 is selected from 4,4'-diaminodicyclohexylmethane (CAS: 1761-71-3) and the like.

Preferably, the compound of II-10 is selected from the group consisting of 1,2-diaminoanthraquinone (CAS: 1758-68-5), 1,5-diaminoanthraquinone (CAS: 129-44-2), 1, 4-Diaminoanthraquinone (CAS: 128-95-0), etc.

According to embodiments of the present invention, compounds of formula II also include substances in which there are two or more structural units represented by formulas II-1 to II-10, such as 4,4'-binaphthylamine (CAS: 481-91-4), 3,3'-dimethylbinaphthylamine (CAS: 13138-48-2), etc.

According to an embodiment of the present invention, the compound of formula II also includes polymers containing multiple amino groups in the structure, such as: polyvinylamine (CAS: 49553-92-6), polyethyleneimine (CAS: 9002-98-6 )wait.

According to embodiments of the present invention, compounds of formula II also include: Direct Brown 44 (CAS: 6252-62-6), Direct Blue 1 (CAS: 2610-05-1), Methylcobalamin (CAS: 13422-55- 4), Vitamin B ₁₂ (CAS: 68-19-9), ethidium bromide (CAS: 1239-45-8), 3,5-diamino-1,2,4-triazole (CAS: 1455 -77-2), new fuchsin (CAS: 3248-91-7), parafuchsin base (CAS: 467-62-9), parafuchsin and its salts, antipartine (CAS: 37691-11- 5), asparagine), lysine and its salts, hydroxylysine, cystine and its salts, arginine and its salts, ornithine and its salts, selenocystamine and its salts, 2 , 7-diaminofluorene (CAS: 525-64-4), Congo red (CAS: 573-58-0), valacyclovir hydrochloride (CAS: 136489-37-7), thymopentin (CAS: 69558 -55-0), 4′,4″(5″)-diaminodibenzo-15-crown-5 (CAS:245086-08-2), adenosylcobalamin (CAS:13870-90-1) , (Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamide (CAS:1450758-21-0), polymyxin B (CAS:1404-26-8 ) and its salts, bacitracin (CAS: 1405-87-4) and its salts, amikacin (CAS: 37517-28-5), pingyangmycin hydrochloride (CAS: 55658-47-4), sulfate chain Mycin (CAS: 3810-74-0), neomycin sulfate (CAS: 1405-10-3), bleomycin (CAS: 11056-06-7), kanamycin, mitomycin ( CAS:50-07-7), Sisomicin sulfate (CAS:53179-09-2), Gentamicin sulfate, Isopamicin sulfate (CAS:67814-76-0), Capreomycin sulfate (CAS:67814-76-0) CAS:1405-37-4), melanostreptin (CAS:3930-19-6), 5,6-diamino-2,4-dihydroxypyrimidine sulfate dihydrate (CAS:63981-35-1 ), famotidine, 4',6-diamidino-2-phenylindole dihydrochloride, etc.

Preferably, the film-forming monomer containing at least two amino groups is selected from arginine, polyethylenimine (PEI), kanamycin, 2-fluoro-p-phenylenediamine, 2,3-dimethyl -p-phenylenediamine, diethylenetriamine, lysine, 3-nitro-1,2-phenylenediamine, p-phenylenediamine, 2,3-diaminonaphthalene, proflavin, 2,3-diamine Aminopyridine, 2,3-diethyl-p-phenylenediamine, 2,4-diaminobenzenesulfonic acid, parafuchsine, 1,6-hexanediamine, ethidium bromide, 2-hydroxymethylp-phenylene Diamine, 2-hydroxyethoxy-p-phenylenediamine, 5,6-diamino-1,3-dimethyluracil, melamine, m-phenylenediamine, 3,4-diaminotoluene, spermine, 4,4'-diaminobenzyl, 3,4-diaminobenzoic acid, 2,4-diaminoanisole, o-toluidine, 4,4'-diaminodicyclohexylmethane, 4,4' -Binaphthylamine, urea, guanidine, 6-hydroxy-2,4,5-triaminopyrimidine, 1,2-diaminocyclohexane, 2,4-diamino-6-methyl-1,3,5 -At least one of triazine, 4,4'-diaminodiphenyl sulfone, 1,4-diaminanthraquinone, and 2,4,6-triaminopyrimidine.

According to an embodiment of the present invention, when the catalyst is at least one of laccase, bilirubin oxidase and peroxidase, the film-forming monomer containing phenolic hydroxyl groups and the film-forming monomer containing at least two amino groups The combination of membrane monomers can be: combination one: a combination of a compound represented by formula I and a compound represented by formula II when there is at least one phenolic hydroxyl group in the structure of R ₁ and/or A ₁ ; or combination two: formula I-13 The combination of the compound shown and the compound represented by formula II-1 when D ₂ is CH substituted by R ₂ and there are at least two amino groups in R ₂ , R ₂ ′, and R ₂ ″.

According to an embodiment of the present invention, when the catalyst is a monophenol monooxidase, the combination of the film-forming monomer containing phenolic hydroxyl groups and the film-forming monomer containing at least two amino groups can be: Formula I A combination of a compound and a compound of formula II.

According to an embodiment of the present invention, when the catalyst is catechol oxidase, the combination of the film-forming monomer containing phenolic hydroxyl groups and the film-forming monomer containing at least two amino groups may be: R ₁ is located adjacent A combination of a compound represented by formula I and a compound represented by formula II when the phenolic hydroxyl group is located at the position of the compound.

According to an embodiment of the present invention, the film-forming monomers containing phenolic hydroxyl groups and the film-forming monomers containing at least two amino groups may be one, two or more in the reaction system.

According to embodiments of the present invention, at least one of an enzyme enhancer, hydrogen peroxide, metal ions, and buffer ion pairs may be added to the enzymatic reaction;

Preferably, the enhancers of the enzyme include α-rythloic acid, β-ryloic acid, γ-ryloic acid, cinnamic acid, 4-hydroxycinnamic acid, needleleaf alcohol, ethyl vanillin, caffeic acid, Ferulic acid, 2,4-pentanedione, 4,4'-dihydroxybenzophenone, benzoic acid, sodium benzoate, 3-hydroxybenzoic acid, 4-hydroxybenzoic acid, 3-hydroxyanthranilic acid, 3-hydroxy-2-aminobenzoic acid, 4-amino-3-hydroxybenzoic acid, 2,3-hydroxybenzoic acid, 3,4-dihydroxybenzoic acid, dimethoxybenzoic acid, 2,4,6- Trihydroxybenzoic acid, p-hydroxyphenylacetic acid, salicylic acid, salicylate, 4-amino-salicylic acid, pyruvic acid, pyruvate, nicotinic acid, ascorbic acid, ascorbate, guanidine, cyanuric acid, imidazole, 2,6-dimethylphenol, 2,4,6-trimethylphenol, 2-acetamidophenol, p-coumaric acid, 7-hydroxycoumarin, sinapinic acid, syringaldehyde, syringaldehyde, clove Acid, methyl syringate, ethyl syringate, propyl syringate, butyl syringate, hexyl syringate, octyl syringate, vanillic acid, isovillic acid, vanillin, vanillin azide, acetosyringone, acetyl Vanillone, vanillyl alcohol, homovanillic acid, catechol, epicatechin, naringin, tyrosine, 2-thiouracil, 3-(4-hydroxy-3,5-dimethoxybenzene ethyl acrylate, quercetin, 2,2'-azino-bis-(3-ethylbenzodihydrothiazoline-6-sulfonic acid) diammonium salt (2,2'-Azino-bis( 3-ethylbenzothiazoline-6-sulfonic acid)diammonium salt, ABTS), hydroxydiphenylethanol, dimethoxybenzyl alcohol, N-hydroxyphthalimide, N-acetyl-N-phenylhydroxylamine, N-benzylidene-benzylamine, N-hydroxy-N-acetylanilide, N-hydroxybutyroanilide, 4,4'-dimethoxy-N-methyl-diphenylamine, 4,4' - Diaminodiphenylamine, triphenylamine, benzidine, N-benzylidene-4-benzidine, 3,3'-dimethylbenzidine, 3,3'-dimethoxybenzidine, 3,3' ,5,5'-tetramethylbenzidine, 4,4'-dihydroxybiphenyl, 4'-hydroxy-4-biphenylcarboxylic acid, phenol red, 2,2',6,6'-tetramethyl Piridine oxide, 1-(3'-sulfophenyl)-3-methyl-5-pyrazolone, uric acid, p-hydroxybenzene sulfonate, 1,5-diaminonaphthalene, 7-methyl Oxy-2-naphthol, 6-hydroxy-2-naphthoic acid, 7-amino-2-naphthalenesulfonic acid, 5-amino-2-naphthalenesulfonic acid, 7-hydroxy-1,2-naphthoimidazole, 1 -Hydroxybenzotriazole, 10-methylphenoxazine, 10-(2-hydroxyethyl)phenoxazine, 10-phenoxazinepropionic acid, phenothiazine-10-propionic acid, 10-(2- Pyrrolidylethyl)phenothiazine, 10-allylphenothiazine, 10-(3-hydroxypropyl)phenothiazine, 2-acetyl-10-methylphenothiazine, 10-ethyl-4 - Phenothiazine carboxylic acid, 10-ethylphenothiazine, 10-propylphenothiazine, 10-isopropylphenothiazine, 10-phenothiazinepropionic acid methyl ester, 10-methylphenothiazine, 10-phenothiazine-propionic acid, thiophenazine, phenothiazine-10-propionic acid (PPT), N-hydroxysuccinimide-10-phenothiazine-propionic acid, 10-(3 -(4-Methyl-1-piperazinyl)propylphenothiazine, promazine, chlorpromazine, 3-hydroxy-1,2,3-benzotriazin-4(3H)-one, N- (4-cyanophenyl)acetoxyxamic acid, iminostilbene, 4-amino-4'-methoxystilbene, 4,4'-diaminostilbene-2,2'-disulfonic acid, 2,7 -Diaminofluorene, 2-(p-aminophenyl)-6-methylbenzothiazole-7-sulfonic acid, N-(4-(dimethylamino)benzylidene)-p-anisaldehyde, 3-methyl -2-Benzothiazolinone (4-(dimethylamino)benzylidene)hydrazone, N-hydroxyacetylide, hardwood black liquor, softwood black liquor, ligno-organosolv, lignosulfonic acid Salt, and mixtures thereof.

According to the embodiment of the present invention, the dosage of the catalyst in the reaction system is 0.01 U/L to 600 U/L based on enzyme activity, for example, 0.5 U/L to 200 U/L, and examples are 1 U/L and 2 U/L. , 6U/L, 12U/L, 24U/L, 40U/L, 60U/L.

According to the embodiment of the present invention, the mass concentration of the film-forming monomer in the reaction system can be 0.002-380g/L, for example, 0.1-20g/L, and examples are 0.5g/L, 0.8g/L, and 1g. /L, 2g/L, 3g/L, 4g/L.

According to the embodiment of the present invention, the film forming temperature may be 4-90°C, for example, 10°C-60°C, and exemplarily 20°C, 30°C, 35°C, 37°C, 40°C, 45°C, 50°C ,55℃;

According to the embodiment of the present invention, the film forming time can be 0.01 to 72h, for example, 2 to 48h, and is exemplarily 3h, 5h, 6h, 8h, 10h, 12h, 20h, 24h, 36h;

According to embodiments of the present invention, the pH of the aqueous phase may be 2 to 10, such as 3, 4, 5, 6, 6.5, 7, or 8.

According to an embodiment of the present invention, when the catalyst is laccase, bilirubin oxidase, monophenol monooxidase or catechol oxidase, the reaction requires the participation of oxygen. The source of oxygen can be pure oxygen or a mixed gas containing oxygen. Further, the oxygen can come from oxygen in the air, can be oxygen dissolved on the surface of the solution, can be through the flow of the solution, mechanical stirring, oscillation, blower, The air compressor slowly dissolves oxygen-containing gas into the reaction system and provides oxygen to the reaction. Oxygen-rich liquid can be added to the liquid.

For reactions that require hydrogen peroxide, the source of hydrogen peroxide in the solution can be at least one of the following methods: (1) Add hydrogen peroxide solution to the solution; (2) Add an enzyme system that generates hydrogen peroxide to the solution, Such as glucose oxidase and glucose, glucose oxidase oxidizes glucose to produce hydrogen peroxide; galactose oxidase and galactose, galactose oxidase oxidizes galactose to produce hydrogen peroxide; sugar alcohol oxidase and glycerol, sugar alcohol oxidase oxidizes glycerol Generate hydrogen peroxide; amino acid oxidase, flavin adenine (FAD) and amino acids, amino acid oxidase uses FAD as a prosthetic group to oxidize amino acids to generate hydrogen peroxide; L-glutamic acid oxidase and L-glutamic acid, L- Glutamic acid oxidase oxidizes L-glutamic acid to produce hydrogen peroxide; (3) Add a source of hydrogen peroxide to the solution, that is, a compound that produces hydrogen peroxide when dissolved in water or a suitable water-based medium, including but Not limited to metal peroxides, tert-butyl hydroperoxide, cumene hydroperoxide, percarbonate, persulfate, perhydrogen persulfate, perphosphate, peracetic acid, perborate, perbenzoic acid, Peroxyacids, alkyl peroxides, acyl peroxides, peroxyesters, urea peroxides and peroxycarboxylic acids or their salts, supplemented by catalase when necessary.

The interface of the two phases can be an upper layer of oil that is liquid at room temperature - a lower layer of water phase (reaction solvent) interface, an upper layer of water phase (reaction solvent) - a lower layer of oil that is liquid at room temperature (such as carbon tetrachloride, carbon disulfide, butyl palmitate). ester) interface, upper water phase (reaction solvent)-lower oil (such as paraffin, beeswax, insect wax, spermaceti, cocoa butter, coconut oil, beef tallow, sheep fat and other animal fats) interface that is solid at the set temperature, The upper water phase (reaction solvent)-lower liquid metal (such as gallium, mercury, low melting point alloy) interface, the interface between the water phase (reaction solvent) droplets suspended in oil and the oil, the interface between the water phase (reaction solvent) droplets suspended in the water phase (reaction solvent) The interface between the oil droplet and the water phase.

The present invention also provides copolymer particles or copolymer films prepared by the synthesis method.

According to the embodiment of the present invention, the copolymer membrane includes the following types: according to the phase where the catalyst and the film-forming monomer are located, it can be divided into the following three types: (1) The catalyst and the film-forming monomer are all dissolved in the water phase The film formed; (2) the catalyst and at least one film-forming monomer are dissolved in the water phase, and the film formed by another or more film-forming monomers in the oil phase; (3) the catalyst is dissolved in the water phase, and the film-forming monomer is The body dissolves in the oil phase to form a film. According to the interface forming the film, it can be divided into the following types: the film at the interface between the upper layer of oil that is liquid at room temperature and the lower layer of water phase, the film at the interface between the upper layer of water phase and the lower layer of oil that is liquid at room temperature, and the film between the upper layer of water phase and the lower layer of liquid metal. Between the membrane and water phase at the interface - the oil that is solid at the set temperature (the oil phase that is solid at the set temperature such as paraffin, beeswax, insect wax, spermaceti, cocoa butter, coconut oil, tallow, etc.) Interface film, oil-water interface film of water-in-oil emulsion, water-oil interfacial film of oil-in-water emulsion.

The present invention also provides applications of the copolymer particles or copolymer films, such as applications in preparing multilayer films, preparing vesicles, preparing conductive films, constructing supramolecular functional materials; and applications in reducing metals and coatings; Application in secondary reactions, such as further modification of copolymer membranes;

Preferably, the modification introduces sorbitol, quaternary ammonium salt, and dye molecules on the surface of the copolymer film.

The copolymer film of the present invention also includes vesicles. Vesicles of the present invention are a type of assembly with a closed structure surrounded by a copolymer membrane.

The invention also provides a method for preparing the vesicles, which includes the following steps: (1) Dissolving the catalyst and the film-forming monomer in the water phase and adding them to the oil-in-water emulsion to obtain vesicles with the copolymer film on the surface. . (2) Dissolve the catalyst and the film-forming monomer in the water phase, pour it into the oil, and stir to obtain a water-in-oil emulsion. Let the mixture react for a period of time to obtain vesicles with the copolymer on the surface. (3) Dissolve the catalyst and a water-soluble film-forming monomer in the water phase, dissolve another oil-soluble monomer in the oil, prepare an emulsion, and react for a period of time to obtain the vesicle wall of the copolymer material film.

The selection of film-forming monomers has a significant impact on the chemical composition, molecular structure, physical and chemical properties and performance of the copolymer film. Therefore, copolymer films can be designed at the molecular level by selecting appropriate film-forming monomers and their ratios. chemical composition, molecular structure, physical and chemical properties and performance, etc. For example: General organic polymers are good insulators, and choose suitable film-forming monomers, such as hydroquinone, ferulic acid, catechol, p-phenylenediamine, benzidine, etc. with conjugated π bonds. The film-forming monomer enables highly delocalized π electrons in the structure of the copolymer film, which can prepare films with conductive properties and can be used for organic polymer full-color flat display materials and devices, optical limiting materials, and sensor sensing. Materials, electrode materials, microwave absorbing materials.

The present invention also provides a multilayer film including at least one layer of the copolymer film.

According to the embodiment of the present invention, in the preparation method, the thickness of the membrane can also be increased by supplementing the raw materials for the enzymatic reaction and/or replacing the fresh enzymatic reaction system.

Those skilled in the art will appreciate that the type and amount of enzymes, the type and concentration of film-forming monomers, pH, temperature, the type and concentration of ion pairs in the buffer, metal ions and their concentrations, oxidants (such as The concentration of dissolved oxygen, hydrogen peroxide), the duration of the enzymatic reaction, enzyme enhancers, inhibitors of enzyme activity, whether it contains organic solvents and the type and concentration of organic solvents, the composition of the liquid medium that constitutes the interface, whether it is updated The reaction solution, whether and how to terminate the reaction, etc., can easily control the reaction rate, the thickness of the membrane and the reactive functional groups on the membrane surface. The thickness of the membrane, and the thickness of the deposits/conjugates on the membrane, can be controlled at the nanometer scale, respectively.

According to the principle of the present invention, the reaction system can use any buffer, pH, ion, and any combination thereof that can satisfy the copolymerization reaction. Other aspects of the enzymatic reaction conditions of the present invention can be optimized through routine experimentation. For example, pH and temperature are non-limiting examples of factors that can be optimized.

By selecting appropriate film-forming monomers, copolymer films that expose specific reactive functional groups can be designed and prepared at the molecular level, that is, the remaining reactive and affinity groups on the copolymer film obtained after the polymerization of the monomers. , the group can be: halogen, unsubstituted or halogen, OH, COOH, NH ₂ , CN, SH, NO ₂ , alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl , aryl, heteroaryl substituted following groups: OH, COOH, NH ₂ , SH, alkyl, alkoxy, alkenyl, alkynyl, aryl, sulfone, sulfonic acid group, amide group, acid halide base, phosphoryl group, azo group. The active groups on the copolymer membrane can introduce functional molecules and/or atoms and/or ions to the surface of the copolymer membrane through secondary or multiple reactions to further functionalize the surface of the copolymer membrane, making the surface of the copolymer membrane It has strong designability and further expands the application scope of the membrane.

According to embodiments of the present invention, the copolymer film can be designed and prepared at the molecular level by changing the type and concentration of film-forming monomers to expose specific reactive functional groups, and the functional groups on the film can undergo chemical or enzymatic reactions. Through chemical or enzymatic reactions, such as group reactions and grafting reactions, the required target molecules/atoms are connected to the membrane to achieve modification and/or modification of the membrane. The group reactions include but are not limited to: Michael-type addition reaction, Schiff base reaction, Friedel-Crafts reaction, Mannich reaction , substitution reaction, addition reaction, polymerization reaction, condensation reaction, oxidation reaction, reduction reaction, esterification reaction, sulfonation reaction, nitration reaction, diazotization reaction, azide reaction, amidation reaction, acylation reaction, alkane reaction Kylation reaction, glycoside-forming reaction, ether-forming reaction, etherification reaction, coupling reaction, complexation reaction, displacement reaction, nucleophilic reaction, nucleophilic elimination reaction, surface atom transfer radical polymerization (SI-ATRP) reaction, reversible Addition-fragmentation chain transfer polymerization (RAFT) reaction.

For example: select appropriate monomers so that the prepared copolymer film contains abundant phenolic hydroxyl groups. Under certain conditions, the phenolic hydroxyl group is oxidized into a quinone structure, which can undergo Michael addition and Schiff base reaction with hydrophilic or hydrophobic organic molecules containing mercapto groups (-SH) and amino groups (-NH ₂ ), thereby converting the functional By introducing molecules to the surface of the copolymer film, self-assembled monolayers can be modified on the surface of the copolymer film.

For example: the phenolic hydroxyl groups in the copolymer film can form coordination bonds with the metal ions in the metal oxide. The copolymer film is immersed in a mixed solution of ammonium hexafluorotitanate ((NH ₄ ) ₂ TiF ₆ ) and boric acid (H ₃ BO ₃ ). The titanium dioxide (TiO ₂ ) generated by the hydrolysis of (NH ₄ ) ₂ TiF ₆ and H ₃ BO ₃ ) The nanoparticles chelate with the copolymer film to form a uniform _TiO2 film on the surface of the copolymer film.

Those skilled in the art will realize that the surface of the copolymer film will contain active groups, so it can combine different types of functional substances and target molecules, such as metal atoms, metal oxide nanoparticles, graphene oxide, functional Living cells, proteins, enzymes, peptides, metal-binding peptides, fluorescent dyes, antibodies, anti-antibodies, haptens, immunogens, DNA, oligonucleotides, growth factors, drug macromolecules, ligands, ligands, amino and / Or thiol and / or aldehyde and / or carboxyl-terminated polyethylene glycol, sugar substances, fatty amines and other substances containing amino and / or thiol and / or aldehyde and / or carboxyl structures, etc.

The copolymer films and multilayer films prepared by the present invention have a rich variety of film-forming monomers to choose from, and the membranes that can be designed and prepared have a rich and diverse composition and molecular structure, which determines the composition, structure, performance and performance of the copolymer film. The service performance is rich and diverse, so you can choose according to the needs of the application.

The copolymer film prepared by selecting appropriate film-forming monomers combined with quaternary ammonium salt molecules can be antibacterial.

The copolymer film of the present invention can construct supramolecular functional materials, assemble two-dimensional and smaller structural materials, construct multi-layer functional polymer materials, and bond or load catalysts to become reactive polymers.

According to embodiments of the present invention, the copolymer membrane can be used to catalyze biological and chemical reactions by combining catalysts, including biological enzymes and chemical catalysts.

Copolymer films prepared by selecting appropriate film-forming monomers can connect dye molecules to express corresponding colors.

According to an embodiment of the present invention, the copolymer film can deposit inorganic substances (silver, hydroxyapatite, calcium carbonate, silicon dioxide, etc.) on its surface to form a nano-layer, from which an organic-inorganic derivative film can be prepared . For example, a metal-modified vesicle is prepared by adding the vesicle to a metal salt solution.

According to embodiments of the present invention, the metal salt includes, but is not limited to, silver nitrate, chloroauric acid, chloroplatinic acid or palladium nitrate.

Metal cations are reduced from the metal salt solution and deposited on the surface of the copolymer film, thereby achieving electroless metallization of the film surface.

In addition, while the copolymer film is being prepared, copolymer particles will be produced in the water phase. The copolymer particles contain high-density active groups such as amino groups, quinone groups, and phenolic hydroxyl groups. On the one hand, they have the characteristics of being reactive. On the other hand, It can be added to plastics as fillers to improve the properties of plastics.

beneficial effects

The copolymer film and its enzymatic self-assembly synthesis method provided by the invention have simple operation steps, no need to isolate air, no external potential, and mild reaction conditions, and can be carried out at normal temperature, normal pressure, neutral or near-neutral pH , requires low energy consumption; the reaction is carried out in liquid, with good chemical uniformity, multi-component uniformity can reach the molecular level, and the composition and structure are uniform; the reaction is easy to control, has good reproducibility, and can be regulated at the molecular level structure. The thickness of the copolymer film and the thickness of the deposits/conjugates on the film can be controlled at the nanometer scale, respectively.

The method for preparing a copolymer film of the present invention has a wide selection range of film-forming monomers and can use many types of film-forming monomers. It can not only polymerize monomers containing catechol structure, but also polymerize resorcinol structure. monomers, monomers with hydroquinone structure, and even compounds with high redox potentials such as bisphenols and biphenyls. On the one hand, different film-forming monomers can be polymerized and cross-linked by laccase to obtain films with different molecular structures. Films with different molecular structures have different properties, performance, and applications. Therefore, the design of this type of film has high With high degree of freedom and strong designability, copolymer membranes with corresponding functions can be selected and prepared according to application requirements. On the other hand, the surface of the copolymer film prepared by selecting suitable film-forming monomers can carry selected active groups to exhibit corresponding activities and properties, and can be produced through secondary/multiple chemical or/and enzymatic reactions. Introduce target molecules and/or atoms into the surface of the membrane to achieve the purpose of functional modification of the membrane surface, making the surface of the polymer membrane highly designable and further expanding the application scope of the membrane.

The copolymer film of the present invention has a high degree of cross-linking, so it has good mechanical properties and good tolerance to water, acid, alkali, organic solvents and heat. Therefore, the vesicles prepared by the present invention have better stability than those prepared with surfactants and proteins.

Based on the above characteristics, the preparation method of the present invention can be easily upgraded from laboratory to industrial mass production, has a short design time, and is easier to start and stop production. The same production line can quickly and flexibly replace products, and is suitable for small batch production processes and distributed manufacturing. , strong adaptability, high flexibility in product, production planning and scheduling. It is particularly worth pointing out that the basic membrane in the present invention is prepared in an enzymatic manner, many of which are difficult to achieve using chemical methods.

The outstanding beneficial effect of the present invention is that it can artificially control the characteristics of the copolymer film with more degrees of freedom and obtain materials that meet the needs, which makes it have great potential in technical applications.

Definitions and explanations of terms

Unless otherwise stated, the definitions of groups and terms recorded in the specification and claims of this application include their definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, and definitions of specific compounds in the examples. etc., can be arbitrarily combined and combined with each other. Such combinations and combined group definitions and compound structures should fall within the scope described in the specification of this application.

The term "film-forming monomer" refers to an organic compound molecule or molecular group that becomes a copolymer film through a polymerization reaction.

The film-forming monomer containing phenolic hydroxyl groups refers to a class of compounds that have one or more hydroxyl groups directly bonded to an aromatic ring in the same molecule.

The film-forming monomer containing at least two amino groups refers to a type of compound, which consists of substances containing at least two amino groups in the same molecule.

The low melting point alloy refers to a metal alloy material with a melting point between 1°C and 96°C, which is not easily volatile, has stable properties, and does not react with water below 120°C.

The artificial enzyme refers to an enzyme mimic with a specific catalytic function synthesized by organic chemistry and biological methods based on the catalytic mechanism of the enzyme, simulating the biocatalytic function of the natural enzyme, and includes metalloenzymes.

The metalloenzyme refers to a conjugated enzyme containing one or several metal ions as prosthetic groups.

Unless otherwise stated, the numerical ranges stated in the specification and claims are equivalent to recording at least each specific integer value therein. For example, the numerical range "1 to 40" is equivalent to recording each integer value in the numerical range "1 to 10", that is, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, and the numerical range Each integer value in "11~40" is 11, 12, 13, 14, 15,..., 35, 36, 37, 38, 39, 40. It should be understood that when one, two or more are used herein to describe a substituent, "more" shall refer to an integer ≥ 3, such as 3, 4, 5, 6, 7, 8, 9 or 10 . In addition, when certain numerical ranges are defined as "numbers," it should be understood that both endpoints of the range, every integer within the range, and every decimal within the range are recited. For example, "numbers from 0 to 10" should be understood as not only recording every integer from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10, but also recording at least one of the integers respectively. The sum of 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9.

The term "halogen" means fluorine, chlorine, bromine and iodine.

The term "C _1-40 alkyl" is understood to mean a straight-chain or branched saturated monovalent hydrocarbon radical having 1 to 40 carbon atoms. For example, "C _1-10 alkyl" means straight and branched chain alkyl groups having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, "C _1-6 alkyl ” represents straight-chain and branched alkyl groups having 1, 2, 3, 4, 5 or 6 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-Methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl base, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl, etc. or their isomers.

The term "C _2-40 alkenyl" is understood to preferably mean a straight or branched monovalent hydrocarbon radical containing one or more double bonds and having 2 to 40 carbon atoms, preferably "C _2-10 alkenyl" . "C _2-10 alkenyl" is understood to mean preferably a straight or branched monovalent hydrocarbon radical which contains one or more double bonds and has 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, for example, having 2, 3, 4, 5 or 6 carbon atoms (i.e., C _2-6 alkenyl), having 2 or 3 carbon atoms (i.e., C _2-3 alkenyl). It will be understood that where the alkenyl group contains more than one double bond, the double bonds may be separated from each other or conjugated. The alkenyl group is, for example, vinyl, allyl, (E)-2-methylvinyl, (Z)-2-methylvinyl, (E)-but-2-enyl, (Z)- But-2-enyl, (E)-but-1-enyl, (Z)-but-1-enyl, pent-4-enyl, (E)-pent-3-enyl, (Z) -Pent-3-enyl, (E)-pent-2-enyl, (Z)-pent-2-enyl, (E)-pent-1-enyl, (Z)-pent-1-enyl base, hex-5-enyl, (E)-hex-4-enyl, (Z)-hex-4-enyl, (E)-hex-3-enyl, (Z)-hex-3- Alkenyl, (E)-hex-2-enyl, (Z)-hex-2-enyl, (E)-hex-1-enyl, (Z)-hex-1-enyl, isopropenyl , 2-methylprop-2-enyl, 1-methylprop-2-enyl, 2-methylprop-1-enyl, (E)-1-methylprop-1-enyl, ( Z)-1-methylprop-1-enyl, 3-methylbut-3-enyl, 2-methylbut-3-enyl, 1-methylbut-3-enyl, 3-methyl Butylbut-2-enyl, (E)-2-methylbut-2-enyl, (Z)-2-methylbut-2-enyl, (E)-1-methylbut-2- Alkenyl, (Z)-1-methylbut-2-enyl, (E)-3-methylbut-1-enyl, (Z)-3-methylbut-1-enyl, (E) )-2-methylbut-1-enyl, (Z)-2-methylbut-1-enyl, (E)-1-methylbut-1-enyl, (Z)-1-methyl But-1-enyl, 1,1-dimethylprop-2-enyl, 1-ethylprop-1-enyl, 1-propylvinyl, 1-isopropylvinyl.

The term "C _2-40 alkynyl" is understood to mean a straight or branched monovalent hydrocarbon radical containing one or more triple bonds and having 2 to 40 carbon atoms, preferably "C _2-10 alkynyl". The term "C _2-10 alkynyl" is understood to mean preferably a straight or branched monovalent hydrocarbon radical which contains one or more triple bonds and has 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, for example, having 2, 3, 4, 5 or 6 carbon atoms (i.e., "C _2-6 alkynyl"), having 2 or 3 carbon atoms (i.e., "C _2-3 alkynyl" ). The alkynyl group is, for example, ethynyl, prop-1-ynyl, prop-2-ynyl, but-1-ynyl, but-2-ynyl, but-3-ynyl, pent-1-ynyl , Pent-2-ynyl, Pent-3-ynyl, Pent-4-ynyl, Hex-1-ynyl, Hex-2-ynyl, Hex-3-ynyl, Hex-4-ynyl, Hex-5-ynyl, 1-methylprop-2-ynyl, 2-methylbut-3-ynyl, 1-methylbut-3-ynyl, 1-methylbut-2-ynyl , 3-methylbut-1-ynyl, 1-ethylprop-2-ynyl, 3-methylpent-4-ynyl, 2-methylpent-4-ynyl, 1-methylpentyl -4-ynyl, 2-methylpent-3-ynyl, 1-methylpent-3-ynyl, 4-methylpent-2-ynyl, 1-methylpent-2-ynyl, 4-methylpent-1-ynyl, 3-methylpent-1-ynyl, 2-ethylbut-3-ynyl, 1-ethylbut-3-ynyl, 1-ethylbut- 2-ynyl, 1-propylprop-2-ynyl, 1-isopropylprop-2-ynyl, 2,2-dimethylbut-3-ynyl, 1,1-dimethylbutyl -3-alkynyl, 1,1-dimethylbut-2-ynyl or 3,3-dimethylbut-1-ynyl. In particular, the alkynyl group is ethynyl, prop-1-ynyl or prop-2-ynyl.

The term "C _3-40 cycloalkyl" is understood to mean a saturated monovalent monocyclic, bicyclic hydrocarbon ring or bridged cycloalkane having 3 to 40 carbon atoms, preferably "C _3-10 cycloalkyl". The term "C _3-10 cycloalkyl" is understood to mean a saturated monovalent monocyclic, bicyclic hydrocarbon ring or bridged cycloalkane having 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. The C _3-10 cycloalkyl group may be a monocyclic hydrocarbon group, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecanyl, or a bicyclic Hydrocarbyl groups such as decalin ring.

The term "3-20 membered heterocyclyl" means a saturated monovalent monocyclic, bicyclic hydrocarbon ring or bridged cycloalkane, which contains a total number of ring atoms of 1 to 5 heteroatoms independently selected from N, O and S. A non-aromatic cyclic group with 3 to 20 atoms (such as 3, 4, 5, 6, 7, 8, 9, 10, etc.) is preferably a "3 to 10-membered heterocyclic group". The term "3-10 membered heterocyclyl" means a saturated monovalent monocyclic, bicyclic hydrocarbon ring or bridged cycloalkane, which contains 1 to 5, preferably 1 to 3 heteroatoms selected from N, O and S. The heterocyclyl group may be attached to the remainder of the molecule through any of the carbon atoms or a nitrogen atom, if present. In particular, the heterocyclyl group may include, but is not limited to: 4-membered rings, such as azetidinyl, oxetanyl; 5-membered rings, such as tetrahydrofuranyl, dioxolyl, pyrrole Alkyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl; or 6-membered ring, such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl Or trithialkyl; or 7-membered ring, such as diazacycloheptyl. Optionally, the heterocyclyl group may be benzo-fused. The heterocyclyl group may be bicyclic, such as but not limited to a 5,5-membered ring, such as a hexahydrocyclopenta[c]pyrrole-2(1H)-yl ring, or a 5,6-membered bicyclic ring, such as a hexahydropyrrole And [1,2-a]pyrazine-2(1H)-yl ring. The ring containing nitrogen atoms may be partially unsaturated, i.e. it may contain one or more double bonds, such as, but not limited to, 2,5-dihydro-1H-pyrrolyl, 4H-[1,3,4]thiodi oxazinyl, 4,5-dihydrooxazolyl or 4H-[1,4]thiazinyl, or it can be benzo-fused, such as but not limited to dihydroisoquinolinyl. According to the invention, the heterocyclyl group is nonaromatic. When the 3-20-membered heterocyclic group is connected to other groups to form the compound of the present invention, the carbon atom on the 3-20-membered heterocyclic group can be connected to other groups, or it can be a 3-20-membered heterocyclic group. The heterocyclic atoms on the ring are connected to other groups. For example, when the 3- to 20-membered heterocyclic group is selected from piperazinyl, the nitrogen atom on the piperazinyl may be connected to other groups. Or when the 3- to 20-membered heterocyclic group is selected from piperidinyl, the nitrogen atom on the piperidinyl ring and the carbon atom in the para position may be connected to other groups.

The term "C _6-20 aryl" is understood to preferably mean a monovalent aromatic or partially aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring having 6 to 20 carbon atoms, preferably "C _6-14 aryl" . The term "C _6-14 aryl" is understood to mean preferably a monovalent or partially aromatic monocyclic, bicyclic or Tricyclic hydrocarbon rings ("C _6-14 aryl"), especially rings with 6 carbon atoms ("C ₆ aryl"), such as phenyl; or biphenyl, or with 9 carbon atoms a ring ("C ₉ aryl"), such as indanyl or indenyl, or a ring having 10 carbon atoms ("C ₁₀ aryl"), such as tetrahydronaphthyl, dihydronaphthyl or naphthyl, Either a ring with 13 carbon atoms ("C ₁₃ aryl"), such as fluorenyl, or a ring with 14 carbon atoms ("C ₁₄ aryl"), such as anthracenyl. When the C _6-20 aryl group is substituted, it may be mono- or poly-substituted. Moreover, there is no restriction on the substitution position, for example, it may be ortho, para or meta substitution.

The term "5-20 membered heteroaryl" is understood to include monovalent monocyclic, bicyclic or tricyclic aromatic ring systems having 5-20 ring atoms and containing 1-5 independently selected from N, O and S heteroatoms, such as "5-14-membered heteroaryl". The term "5-14 membered heteroaryl" is understood to include monovalent monocyclic, bicyclic or tricyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms, in particular 5 or 6 or 9 or 10 carbon atoms, and it contains from 1 to 5, preferably from 1 to 3 heteroatoms each independently selected from N, O and S and, additionally in each case The following can be benzo-fused. In particular, the heteroaryl group is selected from the group consisting of thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiazolyl Diazolyl, thi-4H-pyrazolyl, etc. and their benzo derivatives, such as benzofuryl, benzothienyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzene Triazolyl, indazolyl, indolyl, isoindolyl, etc.; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, etc., and their benzo derivatives, such as quinoline base, quinazolinyl, isoquinolinyl, etc.; or azocinyl, indolizinyl, purinyl, etc. and their benzo derivatives; or cinolinyl, phthalazinyl, quinazolinyl, quinoxalyl Phinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, etc. When the 5-20-membered heteroaryl group is connected to other groups to form the compound of the present invention, the carbon atom on the 5-20-membered heteroaryl ring can be connected to other groups, or the 5-20-membered heteroaryl ring can be connected to other groups. Heteroatoms on the aryl ring are attached to other groups. When the 5-20 membered heteroaryl group is substituted, it may be mono-substituted or poly-substituted. Moreover, there is no restriction on the substitution position. For example, the hydrogen bonded to the carbon atom on the heteroaryl ring may be substituted, or the hydrogen bonded to the heteroatom on the heteroaryl ring may be substituted.

As used herein, the term "derivative" refers to a chemical substance that is structurally related to another substance that is the "original" substance, which may also be referred to as the "parent" compound. "Derivatives" can be made from a structurally related parent compound in one or more steps.

Unless otherwise stated, heterocyclyl, heteroaryl or heteroarylene includes all possible isomeric forms thereof, such as positional isomers thereof. Therefore, for some illustrative non-limiting examples, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11-, 12 may be included Forms in which -, if present, are substituted at one, two or more positions or bonded to other groups, including pyridin-2-yl, pyridinylene-2-yl, pyridin-3-yl, Pyridin-3-yl, pyridin-4-yl, and pyridin-4-yl; thienyl or thienylene includes thiophene-2-yl, thiophene-2-yl, thiophene-3-yl, and thiophene-3 - base; pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, pyrazol-5-yl.

The term "oxo" means that the carbon atom, nitrogen atom or sulfur atom in the substituent is replaced by an oxygen group (=O) formed by oxidation.

The polyphenol refers to a natural, synthetic and semi-synthetic organic compound characterized by the presence of a large number of repeating phenolic structural units. Polyphenols are usually compounds with a molecular weight of 400 to 4000 Da and more than or equal to 5 phenolic hydroxyl groups.

The term "enzyme enhancer" refers to a diffusible molecule that is activated by an oxidase and diffuses from the active site on the enzyme to a sensitive structure. It is a small organic compound with a low redox potential that can catalyze the oxidation process of laccase. as a carrier of electron transfer.

The multilayer film refers to a film composed of at least one copolymer film. In order to distinguish the initially formed film from a film prepared by continuing to carry out secondary or multiple reactions, the present invention refers to the initially formed copolymer film as a "base film". The base membrane is a copolymer membrane prepared through two or more chemical or/and enzymatic reactions, which is called a "derivatized membrane". A new film layer can be generated through enzymatic polymerization on one or both sides of the base film. The chemical composition and structure of the new film layer can be different from the base film, and the new film layer can continue to be formed outside the base film. The base film and the new film layer together form a "multi-layer film". A new polymer film continues to form on the derivatized film, which is called a "derivatized multilayer film."

Detailed ways

The technical solution of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following examples are only illustrative and explain the present invention and should not be construed as limiting the scope of the present invention. All technologies implemented based on the above contents of the present invention are covered by the scope of protection intended by the present invention.

The polyethyleneimine (PEI) in the present invention has an average relative molecular mass of 600 and is manufactured by Shanghai McLean Biochemical Technology Co., Ltd. Paraffin wax is refined paraffin wax with a melting point of 58°C to 60°C, purchased from Jiangsu Shitai Experimental Instrument Co., Ltd. Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available or can be prepared by known methods.

The definition of laccase activity: oxidation of 1 μmol of 2,2'-azino-bis-(3-ethylbenzodihydrothiazoline-6-sulfonic acid) diammonium salt (2,2'- The amount of enzyme required for Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, ABTS) is 1 enzyme activity unit (U).

The definition of manganese peroxidase enzyme activity: The amount of enzyme required to oxidize 1 μmol of ABTS per minute is one enzyme activity unit (U).

The definition of horseradish peroxidase enzyme activity: The amount of enzyme required to oxidize 1 μmol of ABTS per minute is one enzyme activity unit (U).

The definition of lignin peroxidase enzyme activity: The amount of enzyme required to oxidize 1 μmol of ABTS per minute is one enzyme activity unit (U).

The definition of soybean peroxidase enzyme activity: The amount of enzyme required to oxidize 1 μmol of ABTS per minute is one enzyme activity unit (U).

The definition of chloroperoxidase activity: oxidizes 1 μmol of 2-chloro-5,5-dimethyl-1,3-cyclohexanedione per minute to produce 2,2-dichloro-5,5-dimethyl The amount of enzyme required for -1,3-cyclohexanedione is 1 enzyme activity unit (U).

The definition of monophenol monooxidase enzyme activity: The amount of enzyme required to oxidize 1 μmol of L-DOPA per minute is one unit of enzyme activity (U).

The definition of catechol oxidase enzyme activity: The amount of enzyme required to oxidize 1 μmol of catechol per minute is one unit of enzyme activity (U).

The definition of bilirubin oxidase enzyme activity: The amount of enzyme required to oxidize 1 μmol of ABTS per minute is one unit of enzyme activity (U).

Example 1: Preparation of Laccase

(1) Liquid seed culture medium: corn flour 30g/L, soybean cake flour 15g/L, α-amylase 54U/L, NaH ₂ PO ₄ 2.6g/L, KCl 2.25g/L, MgSO ₄ ·7H ₂ O 1.5 g/L, the rest is water, pH 6.0, 0.1MPa sterilization for 25 minutes.

Fermentation medium: fructose 48g/L, corn flour 12g/L, soy peptone 9g/L, (NH ₄ ) ₂ SO ₄ 0.5g/L, KCl 1g/L, NaH ₂ PO ₄ 1.8g/L, MgSO ₄ ·7H ₂ O 0.25g/L, CuSO ₄ ·5H ₂ O 1mmol/L, VB ₁ 0.03g/L, Tween-800.5g/L, vanillin 1mmol/L, the rest is water, pH 6.5, 0.1MPa sterilization 25 minutes.

(2) Liquid seed culture: Take about 3cm ² of the sloping lawn of Polyporus brumalis (strain preservation number CCTCCNO:M 2020809) and inoculate it into a 500mL conical flask. Each bottle is filled with 150mL of liquid seed culture medium and shaken at 30°C. bed, cultured at 150 rpm for 4 days to obtain seed liquid.

Shake flask fermentation culture: Inoculate the seed liquid into the fermentation medium at an inoculum volume ratio of 6%. The fermentation medium has a liquid volume of 150 mL in a 500 mL triangular flask. Shake culture at 30°C and ferment the first to The shaker speed is 150rpm for 3 days, and the shaker speed after 3 days of fermentation is 200rpm.

(3) Preparation of crude enzyme liquid: The fermentation broth was centrifuged at 10,000 × g for 10 minutes at 4°C, and the supernatant was the crude enzyme liquid of laccase.

(4) Purification of laccase:

Take 100 mL of laccase crude enzyme solution, slowly add ammonium sulfate in stages while stirring, so that the concentration of the ammonium sulfate solution gradually reaches 60%. After sufficient precipitation, centrifuge at 12000×g for 10 min at 4°C, collect the precipitate, and dissolve in Add an appropriate volume of pH7.0, 0.02mol/L citric acid-disodium hydrogen phosphate buffer to obtain a salting-out solution. The salted-out solution was dialyzed overnight in the same buffer, and the dialysate was replaced every 10 hours until the conductivity and pH of the liquid inside and outside the dialysis bag were the same to obtain the laccase dialysate.

Equilibrate the chromatography column of DEAE-Sepharose Fast Flow ion exchange medium with pH 7.0, 0.02mol/L citric acid-disodium hydrogen phosphate buffer (liquid A). After the enzyme solution obtained by dialysis was filtered through a filter membrane with a pore size of 0.22 μm, the sample was loaded at a flow rate of 2 mL/min, and the sample volume was 10 mL. After loading the sample, first use liquid A to balance until the baseline returns to zero, and then use citric acid-disodium hydrogen phosphate buffer (liquid B) containing NaCl 0 to 0.5 mol/L for continuous gradient elution, with a flow rate of 1 mL/min. , record the absorbance at a wavelength of 280 nm, collect active components for later use, and collect 2 mL in each tube. The laccase activity and protein concentration of the collected samples were measured, and the eluates with laccase activity were combined to obtain the laccase purified by the chromatography column (referred to as pure enzyme liquid in the present invention), and placed at -20 Store at ℃ for later use.

Example 2: Preparation of copolymer film by enzymatic polymerization at liquid paraffin-aqueous solution interface

(1) Preparation of Coin-arginine copolymer film catalyzed by laccase:

Dissolve 0.050g Coin, 0.050g arginine, and 5.0mg vanillin in a mixed solvent of 35mL 0.05mol/L sodium succinate-succinic acid buffer solution (pH 2.0) and 15mL acetone, add 7.5U of laccase, Mix thoroughly to obtain laccase catalytic system (water phase). Cover the water phase with 3 mm thick liquid paraffin and let it stand for 3 hours in a 30°C incubator (Zhicheng, ZSD-1090, China). A "Coyne-arginine copolymer film" is formed at the interface between the liquid paraffin and the water phase. ". The transfer process of "Coyne-arginine copolymer membrane" is as follows: use a syringe to suck out most of the liquid paraffin on the upper side of the "Coyne-arginine copolymer membrane" and the water phase on the lower side, and re-inject it into the "Coyne-arginine copolymer membrane" -Inject distilled water into the lower side of the "Coyne-arginine copolymer membrane", and repeat the aspiration-injection-aspiration operation several times; insert the coverslip into the water phase under the membrane, and carefully move it to the "Coyne-arginine copolymer membrane" Gently lift it up from underneath the membrane, use filter paper to absorb the remaining liquid paraffin on the surface of the membrane, and dry it at room temperature.

(2) Preparation of irisin-PEI copolymer film catalyzed by laccase:

Dissolve 0.250g irisin, 0.250g PEI, and 12.5mg ABTS in a mixed solvent of 175mL 0.05mol/L potassium hydrogen phthalate-sodium hydroxide buffer solution (pH 5.0) and 75mL ethanol, add 50U of laccase, Mix well to obtain laccase catalytic system (water phase). Cover the water phase with 5 mm thick liquid paraffin and let it stand for 8 hours in a 60°C oven (Zhicheng, ZFD-5090, China). An "irisin-PEI copolymer film" is formed at the interface between the liquid paraffin and the water phase.

Membrane transfer method: Slowly discharge the water phase at the bottom, let the film fall on the surface of the substrate at the bottom of the container, and remove the upper liquid wax.

(3) Preparation of rhein-2,4-diamino-6-methyl-1,3,5-triazine copolymer film using laccase catalysis:

Dissolve 0.125g rhein, 0.125g 2,4-diamino-6-methyl-1,3,5-triazine, and 20mg syringaldehyde in 175mL 0.05mol/L sodium acetate-acetic acid buffer solution (pH 4.0) and Add 5 U of laccase to 75 mL of methanol mixed solvent and mix well to obtain a laccase catalytic system (water phase). The water phase was covered with 5 mm thick liquid paraffin, and left to react for 10 h in a 50°C oven (Zhicheng, ZFD-5090, China). A "rhein-2,4-diamino-6-methyl-1,3,5-triazine copolymer film" is formed at the interface between liquid paraffin and aqueous phase.

(4) Preparation of 1,6-dihydroxynaphthalene-diethylenetriamine copolymer membrane using manganese peroxidase catalysis:

Dissolve 0.125g 1,6-dihydroxynaphthalene and 0.250g diethylenetriamine in a mixed solvent of 200mL 0.05mol/L disodium hydrogen phosphate-citric acid buffer solution (pH 4.0) and 50mL acetone, and add manganese peroxide Mix 12 U of enzyme, 40 μL of 5% hydrogen peroxide and 3.5 mL of 0.1 mol/L MnSO ₄ to obtain a manganese peroxidase catalytic system (water phase). Use a straw to cover the surface of the water phase with 5 mm thick liquid paraffin, and let it stand for 10 hours in a 37°C incubator (Zhicheng, ZSD-1090, China). During this period, 5% hydrogen peroxide was added to the reaction system several times. 40μL. A "1,6-dihydroxynaphthalene-diethylenetriamine copolymer film" is formed at the interface between liquid paraffin and the water phase.

(5) Preparation of shikonin-lysine copolymer membrane catalyzed by manganese peroxidase:

Dissolve 0.125g shikonin and 0.125g lysine in a mixed solvent of 175mL 0.05mol/L disodium hydrogen phosphate-citric acid buffer solution (pH 3.0) and 75mL ethanol, add 10U manganese peroxidase, 5% Mix 40 μL of hydrogen peroxide and 3.5 mL of 0.1 mol/L MnSO ₄ to obtain a manganese peroxidase catalytic system (aqueous phase). Use a straw to cover the surface of the manganese peroxidase catalytic system with 5 mm thick liquid paraffin, and let it stand for 6 hours in a 45°C oven (Zhicheng, ZFD-5090, China). During this period, 5% peroxide was added to the reaction system several times. Hydrogen, 40 μL each time. A "shikonin-lysine copolymer film" is formed at the interface between liquid paraffin and aqueous phase.

(6) Preparation of 3-methylsalicylic acid-3-nitro-1,2-phenylenediamine copolymer membrane catalyzed by manganese peroxidase:

Dissolve 0.010g 3-methylsalicylic acid and 0.010g 3-nitro-1,2-phenylenediamine in 50mL 0.05mol/L sodium succinate-succinic acid buffer solution (pH 5.0), and add manganese peroxide Mix 0.05U of enzyme, 8μL of 5% hydrogen peroxide and 0.7mL of 0.1mol/LMnSO ₄ to obtain a manganese peroxidase catalytic system (water phase). Use a pipette to cover the surface of the manganese peroxidase catalytic system with 5 mm thick liquid paraffin, and let it stand for 12 hours in a 20°C incubator (Zhicheng, ZSD-1090, China). During this period, 5% of the solution was added to the reaction system several times. Hydrogen peroxide, 8 μL each time. A "3-methylsalicylic acid-3-nitro-1,2-phenylenediamine copolymer film" is formed at the interface between liquid paraffin and aqueous phase.

(7) Preparation of gallan blue-2,4,6-triaminopyrimidine copolymer film catalyzed by manganese peroxidase:

Dissolve 0.250g gallic blue and 0.250g 2,4,6-triaminopyrimidine in 250mL 0.05mol/L potassium hydrogen phthalate-sodium hydroxide buffer solution (pH 5.0), add 10U manganese peroxidase, 5 40 μL of % hydrogen peroxide and 3.5 mL of 0.1 mol/LMnSO ₄ were mixed to obtain a manganese peroxidase catalytic system (aqueous phase). Use a straw to cover the surface of the manganese peroxidase catalytic system with 5 mm thick liquid paraffin, and let it stand for 15 hours in a 40°C oven (Zhicheng, ZFD-5090, China). During this period, 5% paraffin was added to the reaction system several times. Hydrogen oxide, 40 μL each time. A "gallo blue-2,4,6-triaminopyrimidine copolymer film" is formed at the interface between liquid paraffin and aqueous phase.

(8) Preparation of 4,4’-dihydroxybiphenyl-2,3-diaminonaphthalene copolymer membrane catalyzed by horseradish peroxidase:

Dissolve 0.125g 4,4'-dihydroxybiphenyl and 0.125g 2,3-diaminonaphthalene in a mixed solvent of 215mL 0.05mol/L glycine-sodium hydroxide buffer solution (pH 10.0) and 35mL acetone, add spicy Mix 6 U of root peroxidase and 40 μL of 5% hydrogen peroxide to obtain a horseradish peroxidase catalytic system (water phase). Use a straw to cover the surface of the horseradish peroxidase catalytic system with 10 mm thick liquid paraffin, and let it stand for 24 hours in a 40°C oven (Zhicheng, ZFD-5090, China). During this period, 5% paraffin was added to the reaction system several times. Hydrogen oxide, 40 μL each time. A “4,4’-dihydroxybiphenyl-2,3-diaminonaphthalene copolymer film” is formed at the interface between liquid paraffin and aqueous phase.

(9) Preparation of bisphenol A-proflavin copolymer film catalyzed by horseradish peroxidase:

Dissolve 0.250g bisphenol A and 0.250g proflavin in a mixed solvent of 175mL 0.1mol/L Tris-hydrochloric acid buffer solution (pH8.0) and 75mL ethanol, add an appropriate amount of horseradish peroxidase 3U and 5% peroxidase Add 40 μL of hydrogen and mix well to obtain the horseradish peroxidase catalytic system (aqueous phase). Use a straw to cover the surface of the horseradish peroxidase catalytic system with 5 mm thick liquid paraffin, and let it stand for 5 hours in a 45°C oven (Zhicheng, ZFD-5090, China). During this period, 5% paraffin was added to the reaction system several times. Hydrogen oxide, 40 μL each time. A "bisphenol A-proflavin copolymer film" is formed at the interface between liquid paraffin and aqueous phase.

(10) Preparation of 1,8,9-trihydroxyanthracene-4,4'-diaminodiphenyl sulfone copolymer membrane using horseradish peroxidase catalysis:

Dissolve 0.040g 1,8,9-trihydroxyanthracene and 0.060g 4,4'-diaminodiphenyl sulfone in 140mL 0.05mol/L disodium hydrogen phosphate-citric acid buffer solution (pH 6.0) and 60mL ethanol. To the mixed solvent, add 12 U of horseradish peroxidase and 40 μL of 5% hydrogen peroxide, mix well, and obtain a horseradish peroxidase catalytic system (water phase). Use a straw to cover the surface of the horseradish peroxidase catalytic system with 5 mm thick liquid paraffin, and let it stand for 17 hours in a 45°C oven (Zhicheng, ZFD-5090, China). During this period, 5% paraffin was added to the reaction system several times. Hydrogen oxide, 40 μL each time. A "1,8,9-trihydroxyanthracene-4,4'-diaminodiphenyl sulfone copolymer film" is formed at the interface between liquid paraffin and the water phase.

(11) Preparation of 4',6,7-trihydroxyisoflavone-parafuchsin copolymer membrane catalyzed by lignin peroxidase:

Dissolve 0.750g 4',6,7-trihydroxyisoflavone and 1.500g parafuchsin in a mixed solvent of 175mL 0.05mol/L sodium citrate-citric acid buffer solution (pH 5.0) and 75mL ethanol, and add lignin Mix 1.5 U of peroxidase and 40 μL of 5% hydrogen peroxide to obtain a lignin peroxide catalytic system (water phase). Use a straw to cover the surface of the lignin peroxide catalytic system with 5 mm thick liquid paraffin, and let it stand for 12 hours in a 45°C oven (Zhicheng, ZFD-5090, China). During this period, 5% paraffin was added to the reaction system several times. Hydrogen oxide, 40 μL each time. A "4',6,7-trihydroxyisoflavone-parafuchsin copolymer film" is formed at the interface between liquid paraffin and aqueous phase.

(12) Preparation of geranin-1,6-hexanediamine copolymer film catalyzed by lignin peroxidase:

Dissolve 0.025g geranol and 0.050g 1,6-hexanediamine in a mixed solvent of 225mL 0.05mol/L trisodium phosphate-phosphate buffer solution (pH 4.0) and 25mL dimethyl sulfoxide, add lignin and Mix 6 U of oxidase and 40 μL of 5% hydrogen peroxide to obtain a lignin peroxide catalytic system (water phase). Use a straw to cover the surface of the lignin peroxide catalytic system with 5 mm thick liquid paraffin, and let it stand for 8 hours in a 30°C incubator (Zhicheng, ZSD-1090, China). During this period, 5% of the lignin peroxide catalytic system was added to the reaction system several times. Hydrogen peroxide, 40 μL each time. A "geranin-1,6-hexanediamine copolymer film" is formed at the interface between liquid paraffin and aqueous phase.

(13) Preparation of resveratrol-5,6-diamino-1,3-dimethyluracil copolymer film catalyzed by soybean peroxidase:

Dissolve 0.025g resveratrol and 0.025g 5,6-diamino-1,3-dimethyluracil in a mixed solvent of 35mL 0.05mol/L trisodium phosphate-phosphate buffer solution (pH 6.0) and 15mL ethanol. , add 0.6 U of soybean peroxidase and 8 μL of 5% hydrogen peroxide, mix well, and obtain a soybean peroxidase catalytic system (water phase). Use a straw to cover the surface of the soybean peroxidase catalytic system with 5 mm thick liquid paraffin, and let it stand for 5 hours in a 65°C oven (Zhicheng, ZFD-5090, China). During this period, 5% paraffin was added to the reaction system several times. Hydrogen oxide, 8 μL each time. A "resveratrol-5,6-diamino-1,3-dimethyluracil copolymer film" is formed at the interface between liquid paraffin and aqueous phase.

(14) Preparation of esculetin-melamine-1,4-diaminoanthraquinone copolymer membrane using soybean peroxidase catalysis:

Dissolve 0.075g esculetin, 0.125g melamine, and 0.075g 1,4-diaminoanthraquinone in a mixed solvent of 175mL 0.05mol/L trisodium phosphate-phosphate buffer solution (pH 5.0) and 75mL ethanol, add soybean Mix 15 U of oxidase and 40 μL of 5% hydrogen peroxide to obtain a soybean peroxidase catalytic system (water phase). Use a straw to cover the surface of the soybean peroxidase catalytic system with 5 mm thick liquid paraffin, and let it stand for 4 hours in a 75°C oven (Zhicheng, ZFD-5090, China). During this period, 5% paraffin was added to the reaction system several times. Hydrogen oxide, 40 μL each time. The "esculetin-melamine-1,4-diaminoanthraquinone copolymer film" is formed at the interface between liquid paraffin and water phase.

(15) Preparation of ellagic acid-PEI copolymer membrane using chloroperoxidase catalysis:

Dissolve 0.025g ellagic acid and 0.050g PEI in 250mL 0.05mol/L sodium citrate-citric acid buffer solution (pH 3.0), add 11.25U chloroperoxidase and 40μL 5% hydrogen peroxide, and mix well. A chloroperoxidase catalytic system (water phase) was obtained. Use a straw to cover the surface of the chloroperoxidase catalytic system with 5 mm thick liquid paraffin, and let it stand for 4 hours in a 35°C incubator (Zhicheng, ZSD-1090, China). During this period, 5% of chlorine was added to the reaction system several times. Hydrogen peroxide, 40 μL each time. An "ellagic acid-PEI copolymer film" is formed at the interface between liquid paraffin and water phase.

(16) Preparation of Jinsongbiflavonoid-spermine copolymer film catalyzed by chloroperoxidase:

Dissolve 0.050g Jinsongbiflavones and 0.100g spermine in a mixed solvent of 70mL 0.05mol/L trisodium phosphate-phosphate buffer solution (pH 3.0) and 30mL ethanol, add 3U chloroperoxidase and 5% peroxidase Add 16 μL of hydrogen and mix well to obtain a chloroperoxidase catalytic system (aqueous phase). Use a straw to cover the surface of the chloroperoxidase catalytic system with 5 mm thick liquid paraffin, and let it stand for 6 hours in a 40°C oven (Zhicheng, ZFD-5090, China). During this period, 5% paraffin was added to the reaction system several times. Hydrogen oxide, 16 μL each time. A "Golden Pine Diflavonoid-Spermine Copolymer Film" is formed at the interface between liquid paraffin and water phase.

(17) Preparation of narcissus-2,4-diaminoanisole copolymer membrane catalyzed by chloroperoxidase:

Dissolve 0.075g narcissus and 0.125g 2,4-diaminoanisole in a mixed solvent of 200mL 0.05mol/L sodium malonate-malonic acid buffer solution (pH 4.0) and 50mL ethanol, add chlorine peroxide Mix 0.75 U of enzyme and 40 μL of 5% hydrogen peroxide to obtain a chloroperoxidase catalytic system (water phase). Use a straw to cover the surface of the chloroperoxidase catalytic system with 5 mm thick liquid paraffin, and let it stand for 5 hours in a 10°C incubator (Zhicheng, ZSD-1090, China). During this period, 5% of chlorine was added to the reaction system several times. Hydrogen peroxide, 40 μL each time. A "narcissin-2,4-diaminoanisole copolymer film" is formed at the interface between liquid paraffin and aqueous phase.

(18) Preparation of tyrosine-o-toluidine copolymer membrane catalyzed by monophenol monooxidase:

Dissolve 0.075g tyrosine and 0.050g o-toluidine in 250mL 0.05mol/L disodium hydrogen phosphate-potassium dihydrogen phosphate buffer solution (pH 6.5), add 150U of monophenol monooxidase, and mix well to obtain monophenol Monooxygenase catalytic system (aqueous phase). Use a straw to cover the surface of the monophenol monooxidase catalytic system with 5 mm thick liquid paraffin, and let it stand for 36 hours in a 20°C incubator (Zhicheng, ZSD-1090, China). A "tyrosine-o-toluidine copolymer film" is formed at the interface between liquid paraffin and aqueous phase.

(19) Preparation of catechol-urea copolymer film catalyzed by catechol oxidase:

Dissolve 7.500g catechol and 5.000g urea in 250mL 0.05mol/L sodium succinate-succinic acid buffer solution (pH 4.0), add 0.75U catechol oxidase, and mix well to obtain catechol oxidase Catalytic system (aqueous phase). Use a straw to cover the surface of the catechol oxidase catalytic system with 5 mm thick liquid paraffin, and let it stand for 8 hours in a 50°C oven (Zhicheng, ZFD-5090, China). A "catechol-urea copolymer film" is formed at the interface between liquid paraffin and aqueous phase.

(20) Preparation of hesperetin-6-hydroxy-2,4,5-triaminopyrimidine copolymer film catalyzed by bilirubin oxidase:

Dissolve 0.020g hesperetin and 0.020g 6-hydroxy-2,4,5-triaminopyrimidine in a mixed solvent of 140mL 0.05mol/L trisodium phosphate-phosphate buffer solution (pH 5.0) and 60mL ethanol, add bile Add 100 U of red pigment oxidase and mix well to obtain the bilirubin oxidase catalytic system (water phase). Use a straw to cover the surface of the bilirubin oxidase catalytic system with 5 mm thick liquid paraffin, and let it stand for 8 hours in a 55°C oven (Zhicheng, ZFD-5090, China). A "hesperetin-6-hydroxy-2,4,5-triaminopyrimidine copolymer film" is formed at the interface between liquid paraffin and aqueous phase.

Example 3: Preparation of copolymer membrane by enzymatic polymerization at aqueous solution-carbon tetrachloride interface

(1) Preparation of 1,3-dihydroxynaphthalene-kanamycin copolymer membrane using laccase catalysis:

Dissolve 0.25g 1,3-dihydroxynaphthalene and 0.25g kanamycin in 50mL 0.05mol/L sodium acetate-acetic acid buffer solution (pH 4.0), add 0.6U of laccase, and mix well to obtain a laccase catalytic system ( water box). First add 55mm thick CCl ₄ (a liquid phase that is immiscible with water) in the container, and then slowly add the water phase to the container. The water phase floats on the CCl ₄ , and a liquid-liquid interface is formed between the two phases. . The reaction was allowed to stand for 12 h in a 35°C incubator (Zhicheng, ZSD-1090, China). A "1,3-dihydroxynaphthalene-kanamycin copolymer film" (air-liquid interface film) is formed on the upper surface of the water phase (the air-liquid interface between the water phase and air), and at the same time, between the water phase and CCl ₄ The liquid-liquid interface forms a "1,3-dihydroxynaphthalene-kanamycin copolymer film" (liquid-liquid interface film). The process of transferring the two interface films to the coverslip: ① First insert the coverslip into the water phase, carefully move it under the "1,3-dihydroxynaphthalene-kanamycin" at the air-liquid interface and place it Pick it up gently to complete the transfer of the air-liquid interface film. ② Use a syringe to suck out the solution on the upper side of the "1,3-dihydroxynaphthalene-kanamycin copolymer membrane" at the liquid-liquid interface, and re-inject distilled water on the upper side of the membrane. Repeat the aspiration-injection- Aspirate the liquid several times, insert another cover slip into CCl ₄ , carefully move it under the liquid-liquid interface film, gently pick up the liquid-liquid interface film, and spread the liquid-liquid interface film flatly on the coverslip. Use filter paper to absorb the remaining CCl ₄ on the lower side of the membrane and dry it at room temperature.

(2) Preparation of ferulic acid-2,3-dimethyl-p-phenylenediamine copolymer membrane using laccase catalysis:

Dissolve 0.125g ferulic acid and 0.125g 2,3-dimethyl-p-phenylenediamine in 250mL 0.05mol/L phthalic acid-hydrochloride buffer (pH 3.0), add 0.5U laccase, and mix well. A laccase catalytic system (water phase) was obtained. First add 5 mm thick CCl ₄ (a liquid phase that is immiscible with water) in the container, and then slowly add the water phase to the container. The water phase floats on the CCl ₄ , and a liquid is formed between the water phase and CCl ₄ . -liquid interface. Let the reaction stand at room temperature for 2 hours. A "ferulic acid-2,3-dimethyl-p-phenylenediamine copolymer film" is formed on the upper surface of the water phase (the gas-liquid interface between the water phase and the air), and at the same time, in the liquid of the water and CCl ₄ phases -The liquid interface forms a "ferulic acid-2,3-dimethyl-p-phenylenediamine copolymer film".

(3) Preparation of p-coumaric acid-p-phenylenediamine copolymer membrane catalyzed by manganese peroxidase:

Dissolve 0.125g p-coumaric acid and 0.25g p-phenylenediamine in 250mL 0.05mol/L tartaric acid-sodium tartrate buffer solution (pH 5.0), add 0.125U manganese peroxidase, 40μL 5% hydrogen peroxide and 0.1 mol/LMnSO ₄ 3.5mL, mix well to obtain a manganese peroxidase catalytic system (aqueous phase). First add 35mm thick CCl ₄ (a liquid phase that is immiscible with water) in the container, and then slowly add the water phase to the container. The water phase floats on the CCl ₄ , and a liquid is formed between the water phase and CCl ₄ . -liquid interface. The reaction was allowed to stand for 36 hours in a 10°C incubator (Zhicheng, ZSD-1090, China). During this period, 5% hydrogen peroxide was added to the water phase several times, 40 μL each time. The upper surface of the water phase (the gas-liquid interface between the water phase and the air) forms a "p-coumaric acid-p-phenylenediamine film", and at the same time, a "p-coumaric acid-p-phenylenediamine film" is formed at the liquid-liquid interface of water and CCl ₄ . Diamine copolymer membrane".

(4) Preparation of 3-methoxysalicylic acid-2,3-diethyl-p-phenylenediamine copolymer membrane using horseradish peroxidase catalysis:

Dissolve 0.125g 3-methoxysalicylic acid and 0.125g 2,3-diethyl-p-phenylenediamine in 250mL 0.05mol/L dipotassium hydrogen phosphate-sodium hydroxide buffer (pH 6.5), add spicy Mix 1.5 U of root peroxidase and 40 μL of 5% hydrogen peroxide to obtain a horseradish peroxidase catalytic system (water phase). First add 25mm thick CCl ₄ (a liquid phase that is immiscible with water) in the container, and then slowly add the water phase to the container. The water phase floats on the CCl ₄ , and a liquid is formed between the water phase and CCl ₄ . -liquid interface. The reaction was allowed to stand for 8 hours in a 55°C oven (Zhicheng, ZFD-5090, China). During this period, 5% hydrogen peroxide was added to the water phase several times, 40 μL each time. The upper surface of the water phase (the gas-liquid interface between the water phase and the air) forms "3-methoxysalicylic acid-2,3-diethyl-p-phenylenediamine", and at the _same time in the liquid- "3-Methoxysalicylic acid-2,3-diethyl-p-phenylenediamine" is formed at the liquid interface.

(5) Preparation of 3,5-diiodosalicylic acid-2-hydroxymethyl p-phenylenediamine copolymer membrane using lignin peroxidase catalysis:

Dissolve 0.050g 3,5-diiodosalicylic acid and 0.100g 2-hydroxymethyl-p-phenylenediamine in 250mL 0.05mol/L tartaric acid-sodium tartrate buffer solution (pH 3.0), and add lignin peroxidase 0.25 U and 40 μL of 5% hydrogen peroxide were mixed to obtain a lignin peroxide catalytic system (water phase). First add 25mm thick CCl ₄ (a liquid phase that is immiscible with water) in the container, and then slowly add the water phase to the container. The water phase floats on the CCl ₄ , forming a layer between the water phase and CCl ₄ . Liquid-liquid interface. The reaction was allowed to stand for 3 hours in a 37°C incubator (Zhicheng, ZSD-1090, China). During this period, 5% hydrogen peroxide was added to the water phase several times, 40 μL each time. The upper surface of the water phase (the gas-liquid interface between the water phase and the air) forms a "3,5-diiodosalicylic acid-2-hydroxymethyl p-phenylenediamine copolymer film". At the same time, in the liquid of water and CCl ₄ -The liquid interface forms a "3,5-diiodosalicylic acid-2-hydroxymethyl p-phenylenediamine copolymer film".

(6) Preparation of gentisic acid-2-hydroxyethoxy-p-phenylenediamine copolymer membrane catalyzed by lignin peroxidase:

Dissolve 0.250g gentisic acid and 0.250g 2-hydroxyethoxy-p-phenylenediamine in 250mL 0.2mol/L disodium hydrogen phosphate-sodium dihydrogen phosphate buffer solution (pH 6.5), and add lignin peroxidase 0.5 U and 40 μL of 5% hydrogen peroxide were mixed to obtain a lignin peroxide catalytic system (aqueous phase). First add 25mm thick CCl ₄ (a liquid phase that is immiscible with water) in the container, and then slowly add the water phase to the container. The water phase floats on the CCl _4. Between the water phase and the liquid phase of CCl ₄ A liquid-liquid interface is formed between them. The reaction was allowed to stand for 36 hours in a 40°C oven (Zhicheng, ZFD-5090, China). During this period, 5% hydrogen peroxide was added to the water phase several times, 40 μL each time. The upper surface of the water phase (the gas-liquid interface between the water phase and the air) forms a "gentisic acid-2-hydroxyethoxy-p-phenylenediamine copolymer film", and at the same time the liquid-liquid interface of water and CCl ₄ forms a " Gentisic acid-2-hydroxyethoxy-p-phenylenediamine copolymer membrane”.

(7) Preparation of p-aminosalicylic acid-m-phenylenediamine copolymer film catalyzed by soybean peroxidase:

Dissolve 0.250g p-aminosalicylic acid and 0.250g m-phenylenediamine in 250mL 0.05mol/L disodium hydrogen phosphate-citric acid buffer solution (pH 7.0), add 0.25U soybean peroxidase and 5% peroxidase Add 40 μL of hydrogen and mix well to obtain the soybean peroxidase catalytic system (water phase). First add 25mm thick CCl ₄ (a liquid phase that is immiscible with water) in the container, and then slowly add the water phase to the container. The water phase floats on the CCl ₄ , forming a layer between the water phase and CCl ₄ . Liquid-liquid interface. The reaction was allowed to stand for 24 h in a 10°C incubator (Zhicheng, ZSD-1090, China). During this period, 5% hydrogen peroxide was added to the water phase several times, 40 μL each time. The upper surface of the water phase (the gas-liquid interface between the water phase and the air) forms a "p-aminosalicylic acid-m-phenylenediamine copolymer film", and at the same time, a "p-aminosalicylic acid-m-phenylenediamine copolymer film" is formed at the liquid-liquid interface of water and CCl ₄ Acid-m-phenylenediamine copolymer membrane”.

(8) Preparation of catechol-caffeic acid-3,4-diaminotoluene copolymer membrane using soybean peroxidase catalysis:

Dissolve 0.025g catechol, 0.025g caffeic acid and 0.050g 3,4-diaminotoluene in 50mL 0.05mol/L tartaric acid-sodium tartrate buffer solution (pH 4.0), add soybean peroxidase 0.15U and 5 % hydrogen peroxide, mix well to obtain soybean peroxidase catalytic system (water phase). First add 25mm thick CCl ₄ (a liquid phase that is immiscible with water) in the container, and then slowly add the water phase to the container. The water phase floats on the CCl ₄ , forming a layer between the water phase and CCl ₄ . Liquid-liquid interface. The reaction was allowed to stand for 6 h in a 20°C incubator (Zhicheng, ZSD-1090, China). During this period, 5% hydrogen peroxide was added to the water phase several times, 8 μL each time. The "catechol-caffeic acid-3,4-diaminotoluene copolymer film" is formed on the upper surface of the water phase (the gas-liquid interface between the water phase and the air), and at the same time, it is formed at the liquid-liquid interface of water and CCl ₄ "Catechol-caffeic acid-3,4-diaminotoluene copolymer film".

(9) Preparation of syringic acid-3,4-diaminobenzoic acid copolymer membrane using chloroperoxidase catalysis:

Dissolve 0.375g syringic acid and 0.200g 3,4-diaminobenzoic acid in 250mL 0.05mol/L sodium acetate-acetic acid buffer solution (pH 3.5), add 1.25U chloroperoxidase and 40μL 5% hydrogen peroxide , mix well to obtain a chlorperoxidase catalytic system (water phase). First add 25mm thick CCl ₄ (a liquid phase that is immiscible with water) in the container, and then slowly add the water phase to the container. The water phase floats on the CCl ₄ , forming a layer between the water phase and CCl ₄ . Liquid-liquid interface. The reaction was allowed to stand for 7 h in a 20°C incubator (Zhicheng, ZSD-1090, China). During this period, 5% hydrogen peroxide was added to the water phase several times, 40 μL each time. The upper surface of the water phase (the gas-liquid interface between the water phase and the air) forms a "syringic acid-3,4-diaminobenzoic acid copolymer film", and at the same time, a "syringic acid-3,4-diaminobenzoic acid copolymer film" is formed at the liquid-liquid interface between water and CCl ₄ . 3,4-diaminobenzoic acid copolymer membrane”.

(10) Preparation of anthocyanin-4,4'-diaminobizyl copolymer membrane using chloroperoxidase catalysis:

Dissolve 0.020g anthocyanin and 0.010g 4,4'-diaminobenzyl in 100mL 0.05mol/L sodium citrate-citric acid buffer solution (pH 4.0), add 1U chloroperoxidase and 5% peroxidase Add 16 μL of hydrogen oxide and mix well to obtain a chloroperoxidase catalytic system (aqueous phase). First add 25mm thick CCl ₄ (a liquid phase that is immiscible with water) in the container, and then slowly add the water phase to the container. The water phase floats on the CCl ₄ , forming a layer between the water phase and CCl ₄ . Liquid-liquid interface. The reaction was allowed to stand for 8 hours in a 25°C incubator (Zhicheng, ZSD-1090, China). During this period, 5% hydrogen peroxide was added to the water phase several times, 16 μL each time. The "anthocyanin-4,4'-diaminobizyl copolymer film" is formed on the upper surface of the water phase (the gas-liquid interface between the water phase and the air), and at the same time, it is formed at the liquid-liquid interface of the water and CCl ₄ phases. "Anthocyanin-4,4'-diaminobizyl copolymer film".

(11) Preparation of 2,6-dihydroxytoluene-4,4'-binaphthylamine copolymer membrane catalyzed by monophenol monooxygenase:

Dissolve 0.050g 2,6-dihydroxytoluene and 0.025g 4,4'-binaphthylamine in 250mL 0.05mol/L potassium hydrogen phthalate-sodium hydroxide buffer solution (pH 5.5), add monophenol monooxidation 2.5U of enzyme, mix well to obtain a monophenol monooxidase catalytic system (water phase). First add 25mm thick CCl ₄ (a liquid phase that is immiscible with water) in the container, and then slowly add the water phase to the container. The water phase floats on the CCl ₄ , forming a layer between the water phase and CCl ₄ . Liquid-liquid interface. The reaction was allowed to stand for 10 h in a 30°C incubator (Zhicheng, ZSD-1090, China). The upper surface of the water phase (the gas-liquid interface between the water phase and the air) forms a "2,6-dihydroxytoluene-4,4'-binaphthylamine copolymer film", and at the same time, the liquid-liquid two-phase water and CCl ₄ phases The interface forms a "2,6-dihydroxytoluene-4,4'-binaphthylamine copolymer film".

(12) Preparation of hydroxytyrosol-guanidine copolymer film catalyzed by catechol oxidase:

Dissolve 1.000g hydroxytyrosol and 1.250g guanidine in 250mL 0.05mol/L disodium hydrogen phosphate-citric acid buffer solution (pH 5.0), add 0.005U catechol oxidase, and mix well to obtain catechol oxidase Catalytic system (aqueous phase). First add 25mm thick CCl ₄ (a liquid phase that is immiscible with water) in the container, and then slowly add the water phase to the container. The water phase floats on the CCl ₄ , forming a layer between the water phase and CCl ₄ . Liquid-liquid interface. The reaction was allowed to stand for 30 h in a 30°C incubator (Zhicheng, ZSD-1090, China). The upper surface of the water phase (the air-liquid interface between the water phase and the air) forms a "hydroxytyrosol-guanidine copolymer film", and at the same time the liquid-liquid interface of the water and CCl _phases forms a "hydroxytyrosol-guanidine copolymer film"".

(13) Preparation of 2,4,6-trihydroxybenzaldehyde-1,2-diaminocyclohexane copolymer membrane catalyzed by bilirubin oxidase:

Dissolve 0.400g 2,4,6-trihydroxybenzaldehyde and 0.400g 1,2-diaminocyclohexane in 200mL 0.05mol/L boric acid-borax buffer solution (pH 8.0), and add 120U of bilirubin oxidase , mix well to obtain the bilirubin oxidase catalytic system (water phase). First add 25mm thick CCl ₄ (a liquid phase that is immiscible with water) in the container, and then slowly add the water phase to the container. The water phase floats on the CCl ₄ , forming a layer between the water phase and CCl ₄ . Liquid-liquid interface. The reaction was allowed to stand for 14 h in a 60°C oven (Zhicheng, ZFD-5090, China). The upper surface of the water phase (the gas-liquid interface between the water phase and the air) forms a "2,4,6-trihydroxybenzaldehyde-1,2-diaminocyclohexane copolymer film". At the same time, both water and CCl ₄ The liquid-liquid interface of the phases forms a "2,4,6-trihydroxybenzaldehyde-1,2-diaminocyclohexane copolymer film".

Example 4: Preparation of copolymer film by enzymatic polymerization at the interface between the upper aqueous phase and the lower liquid gallium

Dissolve 0.125g p-hydroxybenzoic acid and 0.250g 2-fluoro-p-phenylenediamine in 125mL distilled water, adjust the pH to 6.0 with phosphoric acid, add 0.125U laccase, and mix well to obtain a laccase catalytic system (water phase). Put a layer of liquid gallium in a plastic container, and then slowly add the laccase catalytic system (water phase) into the container. The water phase floats on the liquid gallium, forming an upper layer of water between the water phase and the liquid metal gallium. Phase-lower liquid gallium interface. The reaction was allowed to stand for 48 h in a 40°C oven (Zhicheng, ZFD-5090, China). A "p-hydroxybenzoic acid-2-fluoro-p-phenylenediamine copolymer film" is formed at the aqueous solution-liquid gallium interface.

Example 5: Preparation of copolymer membrane by enzymatic polymerization at the upper water phase-lower paraffin interface

Dissolve 0.250g chlorogenic acid and 0.125g 2,3-diaminopyridine in 125mL 0.05mol/L disodium hydrogen phosphate-potassium dihydrogen phosphate buffer solution (pH 7.0), add horseradish peroxidase 0.125U and 5 % hydrogen peroxide, mix well to obtain a horseradish peroxidase catalytic system (aqueous phase). Place the paraffin wax in a container, heat it to melt and level it, and let it sit at room temperature to allow the paraffin wax to cool and solidify into a solid. The horseradish peroxidase catalytic system (aqueous phase) is added to the solid paraffin in the container, and the aqueous phase is located on top of the solidified paraffin. The reaction was carried out with shaking at 100 rpm in a constant-temperature oscillator (Crystal, IS-RDV3, American Jingqi Co., Ltd.) at 25°C for 12 hours. During this period, 5% hydrogen peroxide was added to the water phase several times, 20 μL each time. A "chlorogenic acid-2,3-diaminopyridine copolymer film" is formed on the upper surface of the solid paraffin wax. At the same time, granular "chlorogenic acid-2,3-diaminopyridine copolymer" is produced in the water phase and suspended in the water phase. The aqueous phase above the membrane was discarded and the copolymer membrane was washed with distilled water. Place the container at 70°C, and the paraffin melts into liquid. Insert the PE plate into the liquid paraffin, carefully move it to the bottom of the "chlorogenic acid-2,3-diaminopyridine copolymer membrane" and gently pick up the membrane. Spread flatly on the PE board.

Example 6: Preparation of copolymer membrane by enzymatic polymerization at the upper water phase-lower butter interface

Dissolve 0.300g tannic acid and 0.300g ethidium bromide in 100mL 0.05mol/L sodium malonate-malonic acid buffer solution (pH 6.0), add 0.05U lignin peroxidase and 5% peroxidase Add 15 μL of hydrogen and mix well to obtain a lignin peroxide catalytic system (aqueous phase). Place the butter in a container, heat it to melt and level it, then let it sit at room temperature to allow the butter to cool and solidify. The lignin peroxide catalytic system (aqueous phase) was added to the vessel, the aqueous phase being on top of the solidified tallow. The reaction was carried out with shaking at 100 rpm in a constant-temperature oscillator (Crystal, IS-RDV3, American Jingqi Co., Ltd.) at 25°C for 20 h. During this period, 5% hydrogen peroxide was added to the water phase several times, 15 μL each time. A "tannic acid-ethidium bromide copolymer film" is formed on the upper surface of solid butter. At the same time, granular "tannic acid-ethidium bromide copolymer" is produced in the water phase and suspended in the water phase. The aqueous phase above the membrane was discarded and the copolymer membrane was washed with distilled water. Place the container at 65°C and melt the butter into liquid. Insert the PE board into the liquid butter and carefully move it under the "Tannic Acid-Ethidium Bromide Copolymer Film". Gently pick up the film and spread the film flatly. on the PE board.

Example 7: Preparation of copolymer membrane by enzymatic polymerization at water phase-oil phase interface

Dissolve 0.02g lysine in 50mL 0.05mol/L trisodium phosphate-phosphate buffer solution (pH 5.0), add 2U of laccase, and mix well to obtain the aqueous phase of the laccase catalytic system. Dissolve 0.02g tert-butylhydroquinone in 50mL soybean oil, mix well to obtain the oil phase of the laccase catalytic system, cover it on the water phase, and incubate it in a 30°C incubator (Zhicheng, ZSD-1090, China) and let stand for 5 hours. A "tert-butylhydroquinone-lysine copolymer film" is formed at the interface between the water phase and the oil phase.

Example 8: Preparation of copolymer membrane by enzymatic polymerization at water phase-oil phase interface

Dissolve 0.05g diethylenetriamine in 50mL 0.05mol/L trisodium phosphate-phosphate buffer solution (pH 6.0), add soybean peroxidase 5U and 5% hydrogen peroxide 8μL, mix well, and obtain soybean peroxide The aqueous phase of the enzyme catalyzed system. Slowly cover the water phase with a layer of urushiol with a thickness of more than 1 mm, and let it stand for 5 hours in a 40°C oven (Zhicheng, ZFD-5090, China). During this period, 8 μL of 5% hydrogen peroxide was added to the water phase every 1 hour. . A "urushiol-diethylenetriamine copolymer film" is formed at the interface between urushiol and the water phase.

Example 9: Preparation of multilayer films

Dissolve 0.040g guaiacol and 0.025g 4,4'-diaminodicyclohexylmethane in 50mL 0.05mol/L tartaric acid-sodium tartrate buffer solution (pH 7.0), add 20U of monophenol monooxidase, and mix well. A monophenol monooxidase catalytic system (water phase) was obtained. First, add 55 mm thick CCl ₄ (a liquid phase that is immiscible with water) into the container, and then slowly add the water phase to the container, and the water phase floats on the CCl ₄ . Let the reaction stand for 48 hours in a 25°C incubator (Zhicheng, ZSD-1090, China) to form "guaiacol-4,4'- on the upper surface of the water phase (the gas-liquid interface between the water phase and air). Diaminodicyclohexylmethane copolymer film" (gas-liquid interface film), while forming "guaiacol-4,4'-diaminodicyclohexylmethane copolymer at the liquid-liquid interface of the aqueous phase and CCl ₄ membrane” (liquid-liquid interface membrane). Remove the gas-liquid interface membrane, use a syringe to suck out the solution on the upper side of the "guaiacol-4,4'-diaminodicyclohexylmethane copolymer membrane" at the liquid-liquid interface, and re-inject another solution on the upper side of the membrane. An aqueous phase (dissolve 0.125g gallic acid and 0.125g 2,4-diaminobenzenesulfonic acid in 250mL 0.2mol/L disodium hydrogen phosphate-sodium dihydrogen phosphate buffer solution (pH 7.0), add horseradish peroxidation 0.5 U of enzyme and 40 μL of 5% hydrogen peroxide, mix well), and repeat the aspiration-injection-aspiration operation several times. Place it again in a 40°C oven (Zhicheng, ZFD-5090, China) and let it stand for 20 hours. During this period, 5% hydrogen peroxide is added to the water phase several times, 40 μL each time, on the upper surface of the water phase (water phase). The gas-liquid interface with air) forms a "gallic acid-2,4-diaminobenzenesulfonic acid copolymer film" (gas-liquid interface film), and at the same time the original "guaiacol-4,4'-diaminobenzenesulfonic acid copolymer film" The "cyclohexylmethane copolymer film" liquid-liquid interface film generates a "gallic acid-2,4'-diaminobenzenesulfonic acid copolymer layer" on the upper side of the liquid-liquid interface film, which is different from the original "guaiacol-4,4'-diaminobenzenesulfonic acid copolymer layer". cyclohexylmethane copolymer membrane" combined into a double layer membrane.

Example 10: Enzyme-catalyzed secondary reaction (introduction of sorbitol)

Dissolve 0.1g bisphenol A, 0.1g PEI, and 10mg ABTS in a mixed solvent of 70mL 0.05mol/L trisodium phosphate-phosphate buffer solution (pH 4.0) and 30mL ethanol, add 0.6U laccase, and mix well to obtain lacquer. Enzyme catalytic system (aqueous phase). First, add 55mm thick CCl ₄ (oil phase) to the container, and then slowly add the water phase to the container. The water phase floats on the CCl ₄ and a liquid-liquid interface is formed between the two phases. The reaction was allowed to stand for 10 hours in a 40°C oven (Zhicheng, ZFD-5090, China) to form a "bisphenol A-PEI copolymer film" (gas - liquid interface film), and at the same time, a "bisphenol A-PEI copolymer film" (liquid-liquid interface film) is formed at the liquid-liquid interface of water and CCl ₄ . Remove the gas-liquid interface film. Use a syringe to suck out the solution on the upper side of the "bisphenol A-PEI copolymer membrane" at the liquid-liquid interface, and re-inject another solution (dissolve 0.3g sorbitol in 150mL 0.05mol/L triphosphate) on the upper side of the membrane. Sodium-phosphate buffer solution (pH 5.0), add laccase 10U, 2,2,6,6-Tetramethyl-1-piperidinyloxy (TEMPO) 175mg, Mix well), and repeat the aspiration-injection-aspiration operation several times. Place it again in a 40°C oven (Zhicheng, ZFD-5090, China) for reaction for 24 hours. Use a syringe to suck out the solution on the upper side of the liquid-liquid (CCl ₄ ) interface membrane, re-inject distilled water on the upper side of the liquid-liquid (CCl ₄ ) interface membrane, and repeat the aspiration-injection-aspiration operation several times. Insert the coverslip into CCl ₄ and carefully move it below the liquid-liquid (CCl ₄ ) interface membrane. Gently lift the membrane so that the membrane is spread flat on the coverslip. Use filter paper to absorb the residual CCl ₄ on the underside of the membrane. Allow to dry at room temperature.

The elemental composition of the membrane surface was measured using X-ray Photoelectron Spectroscopy (XPS, ThermoFisher Scientific, Escalab 250xi, USA). After reaction with sorbitol, the N/O ratio on the membrane surface decreased from 0.695 to 0.527. The surface of the bisphenol A-PEI copolymer film without sorbitol contains four elements: C, H, O, and N; while the sorbitol structure contains three elements: C, H, and O, and no N element; therefore, the N on the film surface The decrease in /O ratio proves that the membrane is bound to sorbitol, that is, the surface of the liquid-liquid interface membrane can carry out enzyme-catalyzed secondary reactions.

Example 11: Vesicles and their enzymatic polymerization preparation method

Add 100 μL of oleic acid and 1 mL of 2 mg/mL Hexadecyltrimethyl ammonium bromide (CTAB) solution to the centrifuge tube in sequence, vortex for 5 minutes and then sonicate for 15 minutes to prepare an oil-in-water emulsion. Centrifuge the emulsion at 6000 × g for 5 minutes, remove the lower supernatant with a syringe, and add 1 mL of 0.05 mol/L trisodium phosphate-phosphate buffer solution (pH 5.0) dissolved in 0.05 U laccase, 2 mg hydroquinone, and 2 mg PEI. After oscillation reaction at room temperature for 24 hours in the dark, the outer surface of the oil droplets was wrapped with a hydroquinone-PEI copolymer film, thereby obtaining hydroquinone-PEI copolymer vesicles with oleic acid wrapped inside. The stability of the emulsion substantial improvement.

Place the vesicles prepared above in ethanol and soak them. The ethanol will dissolve the oleic acid and CTAB in the vesicles. Centrifuge at 10000×g for 10 minutes to collect the vesicles. Place them in ethanol again to dissolve the remaining oleic acid and CTAB and centrifuge. Collect vesicles, and so many times. The solvent is evaporated from the vesicles in a negative pressure environment to obtain hollow vesicles.

Hydrophilic or hydrophobic vesicle outer walls can be prepared as needed, and target molecules/atoms can be bonded/deposited on the outer wall of the capsule.

Copolymer vesicles can be used to package cells, organelles, antigens, antibodies, functional polymers, proteins, enzymes, DNA, RNA, polysaccharides, drugs, nutrients, flavors, pesticides, fertilizers, chemicals, fuels, explosives, etc.

Copolymer vesicles can also provide a suitable microenvironment for some chemical reactions and biochemical reactions.

Example 12: Copolymer film reduction of metals

Hydroquinone-PEI vesicles were prepared according to the method in Example 11. The vesicles were immersed in 0.05 mol/L AgNO ₃ solution and left in the dark for 30 minutes. The Ag ⁺ in the AgNO ₃ solution was reduced to Ag nanoparticles and deposited. on the outer surface of the vesicle wall. Centrifuge at 5000 rpm for 5 minutes and wash twice with ultrapure water to obtain vesicles with Ag nanoparticles deposited on the outer surface.

Those skilled in the art will realize that not only Ag ⁺ , but also other metal ions with higher oxidation properties than Ag ⁺ can be reduced by the membrane and deposited on the surface of the membrane, such as reduced gold ions, foil ions, palladium ions, etc. Noble metal ions generate corresponding elements to achieve electroless plating material surface metallization. The material covered with silver particles has excellent antibacterial properties. Palladium, platinum, silver, etc. are important chemical catalysts. Therefore, copolymer films deposited with metals can be used for chemical catalysis. This method can also be used to remove, enrich, and recover these metals from water.

Example 13: Preparation of films with conductive properties

Dissolve 0.25g ferulic acid and 0.25g p-phenylenediamine in 125mL 0.05mol/L trisodium phosphate-phosphate buffer solution (pH 5.0), add 7.5U laccase, and mix well to obtain the laccase catalytic system (water phase) . First add 15mm thick CCl ₄ (oil phase) into the container, and then slowly add the water phase to the container. The water phase floats on CCl ₄ , and a liquid-liquid interface is formed between the water and CCl ₄ phases. The reaction was allowed to stand for 13 hours in a 50°C oven (Zhicheng, ZFD-5090, China). A "ferulic acid-p-phenylenediamine copolymer film" is formed on the upper surface of the water phase (the gas-liquid interface between the water phase and the air), and at the same time, a "ferulic acid-p-phenylenediamine copolymer film" is formed on the liquid-liquid interface of the water and CCl _phases . -P-phenylenediamine copolymer film". Remove the gas-liquid interface film. Use a syringe to suck out the solution on the upper side of the liquid-liquid interface membrane, re-inject distilled water on the upper side of the liquid-liquid (CCl ₄ ) interface membrane, repeat the aspiration-injection-aspiration operation several times, and insert a coverslip into the CCl ₄ , carefully move it below the liquid-liquid (CCl ₄ ) interface membrane, gently lift the membrane, spread the membrane flatly on the coverslip, use filter paper to absorb the residual CCl ₄ on the lower side of the membrane, and dry it at room temperature. The electrical conductivity of the surface of the liquid-liquid (CCl ₄ ) interface film placed on the coverslip was measured to be 1.67 S/m using a resistance tester (Lattice, ST2722, China) by the four-probe method.

Copolymer conductive films can be used for electroluminescence, electrochromism, photoconductivity, electronic switches, all-solid-state batteries, non-linear optical devices, high-density memory materials, flat displays, organic semiconductor devices, molecular wires, light-emitting diodes, anti- Electrostatic materials, electromagnetic shielding materials, photovoltaic cell materials and other fields.

By selecting different film-forming monomers, copolymer films with different conductivities within a certain range can be prepared.

The conductivity of the conductive copolymer film can be improved by doping and composite deposition of metal particles.

Metal ions can be reduced on the surface of the conductive copolymer film, allowing metal elements to be deposited on the surface of the film to improve conductivity.

Comparative example 1:

(1) Dissolve 0.250g bisphenol A and 0.250g proflavin in a mixed solvent of 175mL 0.1mol/L Tris-hydrochloric acid buffer solution (pH 8.5) and 75mL ethanol, and mix well. Use a straw to cover the surface of the above solution (aqueous phase) with 5 mm thick liquid paraffin, and let it stand for 48 hours in a 45°C oven (Zhicheng, ZFD-5090, China). No film-like objects were formed at the interface between liquid paraffin and the water phase, nor were any granular objects produced in the water phase.

(2) Dissolve 0.250g bisphenol A and 0.250g proflavin in a mixed solvent of 175mL 0.1mol/L 10g/L NaOH solution (pH 13.4) and 75mL ethanol, and mix well. Use a straw to cover the surface of the above solution (water phase) with 5 mm thick liquid paraffin, and let it stand for 24 hours in a 45°C oven (Zhicheng, ZFD-5090, China). No film-like objects were formed at the interface between liquid paraffin and the water phase, nor were any granular objects produced in the water phase.

Comparative example 2:

(1) Dissolve 0.125g 4,4'-dihydroxybiphenyl and 0.125g 2,3-diaminonaphthalene in a mixed solvent of 215mL 0.05mol/L glycine-sodium hydroxide buffer solution (pH 8.5) and 35mL acetone , mix well. Use a straw to cover the surface of the above solution (water phase) with 10 mm thick liquid paraffin, and let it stand for 48 hours in a 40°C oven (Zhicheng, ZFD-5090, China). No film-like objects were formed at the interface between liquid paraffin and the water phase, nor were any granular objects produced in the water phase.

(2) Dissolve 0.125g 4,4’-dihydroxybiphenyl and 0.125g 2,3-diaminonaphthalene in a mixed solvent of 215mL 10g/L NaOH solution (pH 13.4) and 35mL acetone, and mix well. Use a straw to cover the surface of the above solution (water phase) with 10 mm thick liquid paraffin, and let it stand for 24 hours in a 40°C oven (Zhicheng, ZFD-5090, China). No film-like objects were formed at the interface between liquid paraffin and the water phase, nor were any granular objects produced in the water phase.

The embodiments of the present invention have been described above. However, the present invention is not limited to the above-described embodiment. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention shall be included in the protection scope of the present invention.

Claims (22)

1. A method for synthesizing a copolymer membrane, characterized in that it includes the following steps: A film-forming monomer containing a phenolic hydroxyl group, a film-forming monomer containing at least two amino groups and a catalyst are dissolved in a water phase and/or a liquid phase that is immiscible with water, and polymerized at the interface of the two phases to form a film to obtain the result. The copolymer film; The catalyst is at least one of a peroxidase and/or an oxidoreductase with laccase activity and/or an artificial enzyme with catalytic activity; The structure of the film-forming monomer containing phenolic hydroxyl groups is shown in Formula I: Wherein, each R ₁ is the same or different and is independently selected from H, halogen, CN, NO ₂ , OH, SH, COOH, the following groups that are unsubstituted or substituted by one, two or more R _a1 : C _1-10 alkyl, C _2-10 alkenyl, C _2-10 alkynyl, C _3-10 cycloalkyl, C _3-10 cycloalkenyl, C _3-10 cycloalkynyl, C _6-14 aryl , 5-14 membered heteroaryl group, 3-10 membered heterocyclic group, -OR _1-2 , -SR _1-3 , -NR _1-4 R _1-5 , -OC(O)R _1-7 , - S(O) ₂ R _1-8 , -OS(O) ₂ R _1-9 , -P(O)R _1-10 R _1-11 , -N＝NR _1-12 ; A ₁ exists or does not exist; when A ₁ exists, it is selected from the group consisting of C _6-14 aryl groups, 5-14 membered heterocyclic groups that are unsubstituted or substituted by one, two or more R _b1 and are connected to the benzene ring. Aryl group, 5-10 membered heterocyclic group; Each R _a1 and R _b1 are the same or different, and are independently selected from H, halogen, CN, OH, SH, NO ₂ , COOH, -OR _1-2 , -SR _1-3 , -NR _1-4 R _{1 -5} , -OC(O)R _1-7 , -S(O) ₂ R _1-8 , -OS(O) ₂ R _1-9 , P(O)R _1-10 R _1-11 , no substitution Or C _1-10 alkyl, C 2-10 alkenyl, C _2-10 alkynyl, _{C 3-10} cycloalkyl, C _3- optionally substituted by one, two or more R _{1-12 10-} ring alkenyl, C _3-10 cycloalkynyl, C _6-14 aryl, 5-14-membered heteroaryl, 3-10-membered heterocyclyl; Each of R _1-2 , R _1-3 , R _1-4 , R _1-5 , R _1-6 , R _1-7 , R _1-8 , R _1-9 , R _1-10 , R _{1- 11.} R _1-12 are the same or different, and are independently selected from H, halogen, CN, OH, SH, NO ₂ , COOH, C _1-10 alkyl, C _2-10 alkenyl, C _2-10 alkynyl , C _3-10 cycloalkyl, C _3-10 cycloalkenyl, C _3-10 cycloalkynyl, C _6-14 aryl, 5-14 membered heteroaryl, 3-10 membered heterocyclyl; The film-forming monomer containing at least two amino groups has a structure represented by formula II: Each R ₂ is the same or different and is independently selected from the group consisting of H, halogen, CN, NO ₂ , NO, OH, SH, COOH, unsubstituted or substituted with one, two or more R _a2 : C _1-10 alkyl, C _2-10 alkenyl, C _2-10 alkynyl, C _3-10 cycloalkyl, C _3-10 cycloalkenyl, C _3-10 cycloalkynyl, 3-14 membered heterocycle Base, -OR _2-2 , -SR _2-3 , -NR _2-4 R _2-5 , -C(O)R _2-6 , -OC(O)R _2-7 , -S(O) ₂ R _2-8 , -OS(O) ₂ R _2-9 , P(O)R _2-10 R _2-11 ; A ₂ is selected from C _1-6 alkylene which is unsubstituted or optionally substituted by one, two or more R _c2 , CH=NN=CH, CH=CH-CO-CH ₂ -CO-CH=CH ; p is an integer from 0 to 12; Each R _a2 and R _c2 are the same or different, and are independently selected from H, halogen, CN, OH, SH, oxo (=O), =NH, NO ₂ , COOH, C _1-10 alkyl, C _{2 -10} alkenyl, C _2-10 alkynyl, C _3-10 cycloalkyl, C _3-10 cycloalkenyl, C _3-10 cycloalkynyl, 3-10 membered heterocyclic group, -OR _2-2 , -SR _2-3 , -NR _2-4 R _2-5 , -C(O)R _2-6 , -OC(O)R _2-7 , -S(O) ₂ R _2-8 , -OS( O) ₂ R _2-9 , P(O)R _2-10 R _2-11 ; Each of R _2-2 , R _2-3 , R _2-4 , R _2-5 , R _2-6 , R _2-7 , R _2-8 , R _2-9 , R _2-10 , R _{2- 11} are the same or different, independently selected from H, halogen, NH ₂ , CN, OH, SH, oxo (=O), NO ₂ , COOH, C _1-10 alkyl, C _2-10 alkenyl, C _2-10 alkynyl group, C _3-10 cycloalkyl group, C _3-10 cycloalkenyl group, C _3-10 cycloalkynyl group, 3-10 membered heterocyclic group. 2. A method for synthesizing a copolymer membrane, characterized in that it includes the following steps: A film-forming monomer containing a phenolic hydroxyl group, a film-forming monomer containing at least two amino groups and a catalyst are dissolved in a water phase and/or a liquid phase that is immiscible with water, and polymerized at the interface of the two phases to form a film to obtain the result. The copolymer film; The catalyst is at least one of a peroxidase and/or an oxidoreductase with laccase activity and/or an artificial enzyme with catalytic activity; The film-forming monomer containing phenolic hydroxyl groups has a structure represented by formulas I-1 to I-8, or I-10 to I-13: Wherein, R ₁ , m and R _c1 have the same definitions as in claim 1; B is selected from N substituted by R ₁ ; D, D ₁ , D ₂ and D ₃ are the same or different, and are independently selected from chemical bonds, unsubstituted Or CH ₂ optionally substituted by one or two R _c1 ; E and E ₁ are the same or different, independently selected from chemical bonds, C _1-6 alkylene, CH=CH, CH=N, N=N or CH=NN=CH; n is an integer from 0 to 5; F ₁ and F ₂ are the same or different, and are independently selected from N, CH or O ⁺ ; R ₁ ', R ₁ ″ and R ₁ have the same definition; The film-forming monomer containing at least two amino groups has a structure represented by formula II-3, II-8 or II-9: _{H2NE3 - NH2} II-3. Wherein, R ₂ and p have the same definitions in claim 1; q is an integer from 0 to 12; R ₂ ' and R ₂ have the same definitions; and at least two of R ₂ and R ₂ ' are amino groups; Each E ₃ and E ₄ are the same or different, and are independently selected from chemical bonds, C _1-6 alkylene groups that are unsubstituted or optionally substituted by one, two or more R _c2 , CH=N, CH= NN=CH, alkylene or alkenyl group containing up to 6 carbon atoms -CO-C _1-6 alkylene group -CO-alkylene or alkenyl group containing up to 6 carbon atoms, containing up to 6 carbon atoms Alkylene or alkenyl group - CO - NH - Alkylene group containing up to 6 carbon atoms - CO - Alkylene or alkenyl group containing up to 6 carbon atoms, Alkylene group containing up to 6 carbon atoms, or Alkenyl-NH-Alkylene having up to 6 carbon atoms-NH-Alkylene or alkenyl having up to 6 carbon atoms, Alkylene or alkenyl having up to 6 carbon atoms-C(O)O - Alkylene group containing up to 6 carbon atoms -C(O)O - Alkylene or alkenyl group containing up to 6 carbon atoms. 3. A method for synthesizing a copolymer membrane, characterized in that it includes the following steps: A film-forming monomer containing a phenolic hydroxyl group, a film-forming monomer containing at least two amino groups and a catalyst are dissolved in a water phase and/or a liquid phase that is immiscible with water, and polymerized at the interface of the two phases to form a film to obtain the result. The copolymer film; The catalyst is at least one of a peroxidase and/or an oxidoreductase with laccase activity and/or an artificial enzyme with catalytic activity; The film-forming monomer containing phenolic hydroxyl groups is selected from the group consisting of 1,3-dihydroxynaphthalene, p-hydroxybenzoic acid, ferulic acid, 1,6-dihydroxynaphthalene, 3-methylsalicylic acid, p-coumaric acid, 4,4'-dihydroxybiphenyl, bisphenol A, chlorogenic acid, 3-methoxysalicylic acid, gallic acid, tannic acid, 3,5-diiodosalicylic acid, gentisic acid, white quinoa Retinol, aescin, para-aminosalicylic acid, caffeic acid, ellagic acid, anthocyanins, syringic acid, tyrosine, guaiacol, 2,6-dihydroxytoluene, catechol, hydroxyl At least one of tyrosol, galloblue, 1,8,9-trihydroxyanthracene, urushiol, and tert-butylhydroquinone; The film-forming monomer containing at least two amino groups is selected from the group consisting of arginine, polyethylenimine (PEI), kanamycin, diethylenetriamine, lysine, 1,6-hexanediamine, and spermine , at least one of 4,4'-diaminodicyclohexylmethane, urea, guanidine, and 1,2-diaminocyclohexane. 4. The synthesis method according to any one of claims 1 to 3, characterized in that after the film-forming monomer containing phenolic hydroxyl groups and the film-forming monomer containing at least two amino groups are dissolved in a solvent, a catalyst is added and mixed. A water phase is obtained, the water phase is brought into contact with a liquid phase that is immiscible with water, and a film is polymerized at the interface of the two phases to obtain the copolymer film. 5. The synthesis method according to any one of claims 1 to 3, characterized in that the film-forming monomer containing phenolic hydroxyl groups is dissolved in a solvent, a catalyst is added and mixed to obtain an aqueous phase, and the composition containing at least two amino groups is The membrane monomer is dissolved in a liquid phase that is immiscible with water, and the water phase is brought into contact with the liquid phase that is immiscible with water, and polymerized to form a film at the interface of the two phases to obtain the copolymer film. 6. The synthesis method according to any one of claims 1 to 3, characterized in that the film-forming monomer containing at least two amino groups is dissolved in a solvent, a catalyst is added and mixed to obtain an aqueous phase, and the composition containing phenolic hydroxyl groups is The membrane monomer is dissolved in a liquid phase that is immiscible with water, and the water phase is brought into contact with the liquid phase that is immiscible with water, and polymerized to form a film at the interface of the two phases to obtain the copolymer film. 7. The synthesis method according to any one of claims 1 to 3, characterized in that the catalyst is dissolved in the water phase, and the film-forming monomer containing phenolic hydroxyl groups and the film-forming monomer containing at least two amino groups are dissolved in A liquid phase that is immiscible with water is brought into contact with the liquid phase that is immiscible with water to cause an enzymatic reaction, and polymerizes to form a film at the interface of the two phases to obtain the copolymer film. 8. The synthetic method according to any one of claims 1-7, wherein the peroxidase is selected from the group consisting of manganese peroxidase, lignin peroxidase, chloroperoxidase and plant peroxidase. At least one of the oxidases; the plant peroxidase is horseradish peroxidase, soybean peroxidase, rice peroxidase, cotton peroxidase, runner bean peroxidase, At least one of chickpea peroxidase, guar peroxidase, and pea peroxidase; The oxidoreductase with laccase activity is selected from at least one of catechol oxidase, monophenol monooxidase and bilirubin oxidase; The artificial enzyme refers to an enzyme mimic with a specific catalytic function that is synthesized using organic chemistry and biological methods based on the catalytic mechanism of the enzyme, simulating the biocatalytic function of the natural enzyme. 9. The method according to claim 8, characterized in that the laccase is a fungal laccase. 10. The method according to claim 9, characterized in that the fungal laccase is Polyporus solani laccase. 11. The method of claim 1, wherein when the catalyst is at least one of laccase, bilirubin oxidase and peroxidase, the film-forming monomer containing phenolic hydroxyl groups The combination with _a film-forming monomer containing at least two amino groups is: a combination of a compound represented by formula I and a compound represented by formula II when there is at least one phenolic hydroxyl group in the structure of R ₁ and/or A 1; When the catalyst is a monophenol monooxidase, the combination of the film-forming monomer containing a phenolic hydroxyl group and the film-forming monomer containing two amino groups is: a combination of a compound represented by formula I and a compound represented by formula II. ; When the catalyst is catechol oxidase, the combination of the film-forming monomer containing phenolic hydroxyl groups and the film-forming monomer containing at least two amino groups is: the formula when R ₁ is a phenolic hydroxyl group located in the ortho position The combination of the compound shown in I and the compound shown in formula II; The film-forming monomers containing phenolic hydroxyl groups and the film-forming monomers containing at least two amino groups are respectively one or more than two in the reaction system. 12. The method according to any one of claims 1 to 3, characterized in that the aqueous phase is an aqueous solution with a water content of not less than 50% in the total mass. 13. The method according to claim 12, characterized in that the aqueous phase is dissolved with film-forming monomers and contains an organic solvent dissolved in water, and the organic solvent is methanol, ethanol, isopropyl alcohol, acetone, formic acid At least one of methyl ester, ethyl acetate, acetonitrile, tetrahydrofuran, N,N-dimethylformamide, 1,4-dioxane, dimethyl sulfoxide, diethylene glycol butyl ether, and diethylene glycol kind. 14. The method according to claim 7, wherein at least one of an enzyme enhancer, hydrogen peroxide, metal ions, and buffer ion pairs is added to the enzymatic reaction. 15. The method of claim 7, wherein the liquid phase that is immiscible with water is selected from the group consisting of oil that is liquid at room temperature, oil that is solid at a set temperature, and liquid metal. 16. The method according to claim 4, characterized in that the solvent is selected from water or a buffer solution, and the buffer solution is sodium acetate-acetic acid buffer solution, disodium hydrogen phosphate-citric acid buffer solution, phthalate Potassium hydrogen formate-sodium hydroxide buffer solution, tartaric acid-sodium tartrate buffer solution, sodium citrate-citric acid buffer solution, trisodium phosphate-phosphate buffer solution, sodium malonate-malonic acid buffer solution, sodium succinate-amber Acid buffer solution, phthalic acid-hydrochloric acid buffer, disodium hydrogen phosphate-sodium dihydrogen phosphate buffer solution, disodium hydrogen phosphate-potassium dihydrogen phosphate buffer solution, dipotassium hydrogen phosphate-sodium hydroxide buffer, Tris- Hydrochloric acid buffer solution, boric acid-borax buffer solution, glycine-sodium hydroxide buffer solution. 17. The method according to any one of claims 1 to 3, characterized in that the dosage of the catalyst in the reaction system is 0.01 U/L to 600 U/L based on enzyme activity. 18. The method according to any one of claims 1 to 3, characterized in that the dosage of the catalyst in the reaction system is 0.5 U/L to 200 U/L based on enzyme activity. 19. The method according to any one of claims 1 to 3, characterized in that the mass concentration of the film-forming monomer in the reaction system is 0.002-380g/L. 20. The method according to any one of claims 1 to 3, characterized in that the temperature of the film-forming reaction is 4 to 90°C. 21. The method according to any one of claims 1 to 3, characterized in that the film-forming reaction time is 0.01 to 72 hours. 22. The method according to any one of claims 1-3, characterized in that the pH of the aqueous phase is 2-10.