CS270956B1 - Column for enzyme transformation of substances - Google Patents
Column for enzyme transformation of substances Download PDFInfo
- Publication number
- CS270956B1 CS270956B1 CS878733A CS873387A CS270956B1 CS 270956 B1 CS270956 B1 CS 270956B1 CS 878733 A CS878733 A CS 878733A CS 873387 A CS873387 A CS 873387A CS 270956 B1 CS270956 B1 CS 270956B1
- Authority
- CS
- Czechoslovakia
- Prior art keywords
- column
- enzyme
- substances
- groove
- capillary
- Prior art date
Links
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 17
- 102000004190 Enzymes Human genes 0.000 title claims abstract description 17
- 239000000126 substance Substances 0.000 title abstract description 9
- 230000009466 transformation Effects 0.000 title 1
- 238000006243 chemical reaction Methods 0.000 claims abstract description 10
- 239000007788 liquid Substances 0.000 claims abstract description 8
- 108010093096 Immobilized Enzymes Proteins 0.000 claims abstract description 3
- 239000000203 mixture Substances 0.000 claims abstract description 3
- 239000012530 fluid Substances 0.000 claims description 2
- 230000000284 resting effect Effects 0.000 claims description 2
- 230000002255 enzymatic effect Effects 0.000 abstract description 4
- 229940088598 enzyme Drugs 0.000 description 12
- 230000000694 effects Effects 0.000 description 2
- 229910001369 Brass Inorganic materials 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 108010015776 Glucose oxidase Proteins 0.000 description 1
- 239000004366 Glucose oxidase Substances 0.000 description 1
- 239000004952 Polyamide Substances 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 239000011942 biocatalyst Substances 0.000 description 1
- 239000010951 brass Substances 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 210000003027 ear inner Anatomy 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229940116332 glucose oxidase Drugs 0.000 description 1
- 235000019420 glucose oxidase Nutrition 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 230000036284 oxygen consumption Effects 0.000 description 1
- 239000006174 pH buffer Substances 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 229920002647 polyamide Polymers 0.000 description 1
- 229920002379 silicone rubber Polymers 0.000 description 1
- 239000004945 silicone rubber Substances 0.000 description 1
- 229910001220 stainless steel Inorganic materials 0.000 description 1
- 239000010935 stainless steel Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
- C12M21/18—Apparatus specially designed for the use of free, immobilized or carrier-bound enzymes
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J16/00—Chemical processes in general for reacting liquids with non- particulate solids, e.g. sheet material; Apparatus specially adapted therefor
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J19/0093—Microreactors, e.g. miniaturised or microfabricated reactors
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/06—Tubular
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/54—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving glucose or galactose
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00781—Aspects relating to microreactors
- B01J2219/00819—Materials of construction
- B01J2219/00824—Ceramic
- B01J2219/00828—Silicon wafers or plates
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00781—Aspects relating to microreactors
- B01J2219/00819—Materials of construction
- B01J2219/00835—Comprising catalytically active material
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Sustainable Development (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Clinical Laboratory Science (AREA)
- Emergency Medicine (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biophysics (AREA)
- Immunology (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
Description
Vynález se týká kolony pro enzymovou přeměnu látek, skládající se z pouzdra kolony, opatřené přívodem a odvodem kapaliny. Kolonu lze využít ve spojení s průtočným detektorem, například voltamperometrickým nebo fotometrickým, ke sledování koncentrace látek, které jsou substráty použitých enzymů či enzymových systémů nebo látek, které ovlivňují enzymové aktivity těchto enzymů.BACKGROUND OF THE INVENTION The present invention relates to a column for enzymatic conversion of substances consisting of a column housing provided with a fluid inlet and outlet. The column can be used in conjunction with a flow detector, for example voltamperometric or photometric, to monitor the concentration of substances that are substrates of the enzymes used or of the enzyme systems or substances that affect the enzymatic activities of these enzymes.
Jsou známy náplňové i kapilární kolony obsahující biokatalyzátor, který katalyzuje přeměnu jedné nebo více látek procházejících kolonou. Jednou z možností tako- r vé katalýzy je enzymová přeměna sledované látky, přičemž tato reakce je doprovázena například spotřebou kyslíku a uvolněním celé řady dalších látek, jejichž koncentrace může být sledována voltamperometricky, fotometrickým detektorem a podobně. Nevýhodou stávajících kapilárních i náplňových kolon je nesnadná výměna náplně těchto kolon a nakonec i často Časově náročná výměna kolon v zařízeních, ke kterým je nutno sáhnout především při ztrátě aktivity enzymu nebo při ucpání kolon.Both packed and capillary columns containing a biocatalyst are known which catalyze the conversion of one or more substances passing through the column. One possibility of such R in the catalysis of the enzyme conversion of substance, which reaction is accompanied by e.g. oxygen consumption and release of many other substances whose concentrations may be monitored voltamperometricky, a photometric detector or the like. The disadvantage of existing capillary and packed columns is the difficulty of replacing the packings of these columns and, in the end, often the time-consuming exchange of columns in equipment, which must be achieved especially in the case of loss of enzyme activity or column clogging.
Uvedené nedostatky odstraňuje kolona pro enzymovou přeměnu látek, sestávající z pouzdra kolony, opatřené přívodem a odvodem kapaliny, podle vynálezu. Jeho podstata spočívá v tom, že kapilární kolona válcového tvaru je opatřena na Čelní ploše spirálově vytvořenou drážkou s imobilizovaným enzymem nebo směsí enzymů, proti níž je upraveno pružné těsnění, opatřené v místě přívodu a odvodu kapaliny otvory, přičemž na kolonu dosedá přítlačná vložka, opřená o víko pouzdra kolony.These drawbacks are overcome by an enzyme conversion column consisting of a column housing provided with a liquid inlet and outlet according to the invention. It consists in that a cylindrical capillary column is provided with a spiral groove on the front surface with an immobilized enzyme or a mixture of enzymes against which a flexible seal is provided, provided with openings at the liquid inlet and outlet, with a pressure pad resting against the column. o Column housing cover.
Vyšší účinek vynálezu spočívá ve snadnosti imobilizace enzymů do drážek vytvořených v kruhové ploše, ve snadné a rychlé výměně labyrintů kapilární kolony, a to i méně kvalifikovanou obsluhou a možností umístění držáku kolony přímo do panelu chemické části přístroje, to je analyzátoru, což je umožněno uložením přívodu a vývodu kolony, situovaných na jedné straně kolonového pouzdra.The higher effect of the invention is the ease of immobilization of enzymes into the grooves formed in the circular surface, the easy and quick replacement of the capillary column labyrinths, even by less qualified operators and the possibility of placing the column holder directly in the chemical panel of the instrument. column inlet and outlet situated on one side of the column housing.
Kolona pro enzymovou přeměnu látek je dále objasněna na příkladu provedení podle připojeného výkresu, na němž je znázorněna kolona ve schematickém nárysu.The enzyme conversion column is further elucidated by way of example with reference to the accompanying drawing, in which the column is shown in schematic elevation.
Kolona sestává ze základního tělesa _1, v němž je umístěna kapilární kolona 5 válcového tvaru, opatřená na čelní ploše drážkou 8, která je vytvořena spirálově vedeným zahloubením od středu až téměř к obvodu kruhové plochy kapilární kolony 5· V drážce 8 je imobilizován enzym nebo směs enzymů. Proti drážce _8 je umístěno pružné těsnění 7, které je opatřeno otvory v místě přívodu 3 a odvodu 4 kapaliny. Kapilární kolona 5 je proti pružnému těsnění 7 dotlačována pomocí přítlačné vložky Д, opírající se o víko 2 pouzdra kolony. ,The column consists of a base body 1, in which a cylindrical capillary column 5 is provided, provided with a groove 8 on its face, formed by a spiral recess from the center to almost the periphery of the circular surface of the capillary column 5. enzymes. Opposite the groove 8 a resilient seal 7 is provided, which is provided with openings at the points of inlet 3 and outlet 4 of the liquid. The capillary column 5 is pressed against the resilient seal 7 by means of a thrust insert Д, supported on the lid 2 of the column housing. ,
S kolonou se pracuje následujícím způsobem. Přívodem^ se přivádí kapalina, která prochází otvorem v pružném těsnění 7 do drážky 8, v níž je imobilizován enzym. Otvor je umístěn ve středu pružného těsnění 7» zde je na počátek spirálové kapilární kolony 5 přiváděn analyzovaný roztok, procházející dále spirálovou kapilární ко- д lonou 5 a opouštějící tuto kolonu až téměř u obvodu kruhové plochy kapilární kolony potvorem v pružném těsnění 7 a odvodem 4 kapaliny.The column is operated as follows. Liquid is supplied through the inlet 6, which passes through an opening in the resilient seal 7 to a groove 8 in which the enzyme is immobilized. The aperture is located in the center of the flexible seal 7 »here the analyte is fed to the beginning of the helical capillary column 5, passing further through the helical capillary column 5 and leaving this column almost to the periphery of the circular surface of the capillary column through the bore in the flexible seal 7 and liquid.
Kolona pro enzymovou přeměnu látek podle vynálezu byla odzkoušena v následujícím konkrétním provedení. Základní těleso l^bylo zhotoveno z organického skla a by, · lo opatřeno kapilárním přívodem 3 a odvodem 4 zhotoveným z nerezavějící oceli. Víko _2 a přítlačná vložka_£byly zhotoveny z mosazi, drážka 8 kapilární kolony 5 byla vysoustružena z polyamidu a jako pružné těsnění 7 byl použit kotouč ze silikonové pryže, do něhož byly vyseknuty v místě přívodu 3 a odvodu 4 otvory o průměru cca 1,5 mm. Do drážky 8 kapilární kolony 5 byla koimobilizována glukozooxidaza a katalaza. Sestavená kolona pro enzymovou přeměnu látek byla testována, ve spojení s přesným diskontinuálním dávkovačem malých množství vzorku, kyslíkovým čidlem a kontinuálním dávkovačem tlumivého roztoku pH, pomocí roztoku glukózy, jejíž koncentrace se pohybovala v koncentračním rozsahu 0 až 20 mM.dm~3. Pro koncentraci 20 mM.dm-3 činila relativní směrodatná odchylka výšek píku + 1,5The enzymatic conversion column of the compounds of the invention was tested in the following specific embodiment. The base body 1 was made of organic glass and would have a capillary inlet 3 and a outlet 4 made of stainless steel. The lid 2 and the pressure pad 6 were made of brass, the groove 8 of the capillary column 5 was turned from polyamide, and a silicone rubber disc was used as a resilient seal 7 and punched about 1.5 in. mm. Glucose oxidase and catalase were co-immobilized into the groove 8 of the capillary column 5. The assembled enzyme conversion column was tested, in conjunction with an accurate batch small batch sampler, an oxygen sensor, and a continuous pH buffer dispenser, with a glucose solution ranging in concentration from 0 to 20 mM.dm- 3 . For a concentration of 20 mM.dm -3 , the relative standard deviation of the peak heights was + 1.5
Claims (1)
Priority Applications (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS878733A CS270956B1 (en) | 1987-12-02 | 1987-12-02 | Column for enzyme transformation of substances |
| DE19893906911 DE3906911A1 (en) | 1987-12-02 | 1989-03-03 | COLUMN FOR ENZYMATIC CONVERSION OF SUBSTANCES |
| FR8903149A FR2644175A1 (en) | 1987-12-02 | 1989-03-10 | COLUMN FOR AN ENZYMATIC CONVERSION OF SUBSTANCES |
| GB8905529A GB2228879A (en) | 1987-12-02 | 1989-03-10 | Device for enzymatic conversion of substances |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS878733A CS270956B1 (en) | 1987-12-02 | 1987-12-02 | Column for enzyme transformation of substances |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CS873387A1 CS873387A1 (en) | 1990-01-12 |
| CS270956B1 true CS270956B1 (en) | 1990-08-14 |
Family
ID=5438313
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CS878733A CS270956B1 (en) | 1987-12-02 | 1987-12-02 | Column for enzyme transformation of substances |
Country Status (4)
| Country | Link |
|---|---|
| CS (1) | CS270956B1 (en) |
| DE (1) | DE3906911A1 (en) |
| FR (1) | FR2644175A1 (en) |
| GB (1) | GB2228879A (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1157738B1 (en) * | 2000-05-21 | 2005-11-30 | CPC Cellular Process Chemistry Systems GmbH | Residence time modul for microreactors |
-
1987
- 1987-12-02 CS CS878733A patent/CS270956B1/en unknown
-
1989
- 1989-03-03 DE DE19893906911 patent/DE3906911A1/en not_active Withdrawn
- 1989-03-10 GB GB8905529A patent/GB2228879A/en not_active Withdrawn
- 1989-03-10 FR FR8903149A patent/FR2644175A1/en not_active Withdrawn
Also Published As
| Publication number | Publication date |
|---|---|
| DE3906911A1 (en) | 1990-10-04 |
| FR2644175A1 (en) | 1990-09-14 |
| GB8905529D0 (en) | 1989-04-19 |
| GB2228879A (en) | 1990-09-12 |
| CS873387A1 (en) | 1990-01-12 |
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