WO2017109897A1 - Procédé de production glycérophospholipide de type éther - Google Patents

Procédé de production glycérophospholipide de type éther Download PDF

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WO2017109897A1
WO2017109897A1 PCT/JP2015/086032 JP2015086032W WO2017109897A1 WO 2017109897 A1 WO2017109897 A1 WO 2017109897A1 JP 2015086032 W JP2015086032 W JP 2015086032W WO 2017109897 A1 WO2017109897 A1 WO 2017109897A1
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Prior art keywords
ether type
type glycerophospholipid
glycerophospholipid
enzyme
scallop
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English (en)
Japanese (ja)
Inventor
康尋 貴家
勝 福井
武彦 藤野
志郎 馬渡
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B&S Corp KK
Institute of Rheological Function of Food Co Ltd
Inst of Rheological Function of Food Co Ltd
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B&S Corp KK
Institute of Rheological Function of Food Co Ltd
Inst of Rheological Function of Food Co Ltd
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Priority to JP2017557588A priority Critical patent/JP7021953B2/ja
Priority to PCT/JP2015/086032 priority patent/WO2017109897A1/fr
Publication of WO2017109897A1 publication Critical patent/WO2017109897A1/fr
Anticipated expiration legal-status Critical
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/06—Phosphorus compounds without P—C bonds
    • C07F9/08—Esters of oxyacids of phosphorus
    • C07F9/09—Esters of phosphoric acids
    • C07F9/10—Phosphatides, e.g. lecithin
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
    • C11B1/00—Production of fats or fatty oils from raw materials
    • C11B1/10—Production of fats or fatty oils from raw materials by extracting
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
    • C11B7/00—Separation of mixtures of fats or fatty oils into their constituents, e.g. saturated oils from unsaturated oils
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P13/00—Preparation of nitrogen-containing organic compounds
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P9/00—Preparation of organic compounds containing a metal or atom other than H, N, C, O, S or halogen

Definitions

  • the present invention relates to a method of producing ether type glycerophospholipid. More specifically, the present invention relates to a method for efficiently producing a highly pure ether type glycerophospholipid by a simple operation.
  • a lipid refers to a substance having a long chain fatty acid or a similar hydrocarbon chain in the molecule and present in the living body or derived from the living body.
  • the lipids can be roughly classified into simple lipids and complex lipids.
  • the simple lipid is composed of carbon (C), hydrogen (H) and oxygen (O) and is generally soluble in acetone, and the simple lipid triacylglycerol is used as a reservoir of energy in adipose tissue in the animal body.
  • a complex lipid is a lipid group that contains phosphoric acid and sugar in the molecule, is composed of a hydrophobic part (fatty acid part) and a hydrophilic part (phosphoric acid and base part), and exhibits amphiphilic property
  • Many are the main constituent lipids of lipid bilayers of cell membranes, and are also involved in signal transmission in the body.
  • the complex lipids are insoluble in acetone while the simple lipids are soluble in acetone.
  • the complex lipid is 1) Glycerophospholipids; These include phosphatidyl choline (also called lecithin), phosphatidyl ethanolamine and the like. 2) sphingophospholipids; Sphingomyelin, ceramide syriatin etc. belongs. 3) glycosphingolipids; These include cerebroside, sulfatide, ganglioside and the like. And 4) glyceroglycolipids; Examples include diacylglycerols present in microorganisms and higher plants, in which various sugars are linked. The sphingophospholipids of 2) and the glycosphingolipids of 3) are collectively called sphingolipids.
  • the glycerophospholipid is a general term for phospholipids having glycerol as a backbone, and is important as a component of biological membranes.
  • the glycerophospholipids can be divided into subclasses of diacyl glycerophospholipids, alkenylacyl glycerophospholipids (plasmogen), and alkyl ether glycerophospholipids.
  • the alkenyl acyl type glycerophospholipid (plasmalogen) and the alkyl ether type phospholipid are collectively referred to as ether type glycerophospholipids because they have an ether bond.
  • plasmalogen having a vinyl ether bond at position 1 (sn-1) of fatty acid is a phospholipid which is characteristically contained in brain neurons, cardiac muscle, skeletal muscle and the like, and is a lipid component attracting attention in recent years.
  • This plasmalogen bears a biological function characterized by the vinyl ether structure in the molecule, and it has an antioxidant property by capturing active oxygen, radicals and metal ions, and exhibits flowability of cell membrane (especially nerve cell synaptic membrane) Have been reported to be involved (Non-Patent Document 1).
  • Non-patent Documents 1 and 2 the brain of Alzheimer's disease has a significantly lower plasmalogen-type phospholipid concentration and a reduction of about 30% as compared to the brain of a healthy adult.
  • Patent Documents 1 to 4 it has been proposed to improve or prevent diseases such as Alzheimer's disease by incorporating plasmalogen or ether type glycerophospholipids in food and drink or medicines.
  • Patent Literatures 1 and 2 disclose an example of a method for producing such ether type glycerophospholipid.
  • Patent Document 5 proposes a method for extracting plasmalogen-containing lipids from marine animals using only a solvent that can also be used in the food and drink field.
  • the extraction method of this plasmalogen-containing lipid is 1) An extraction process of adding a mixed solvent of n-hexane / ethanol / water to a dried product of marine animals and stirring, and collecting a filtrate; 2) A liquid / liquid separation step of adding a mixed solvent of n-hexane / water to the filtrate collected in the above extraction step, stirring it, leaving it still and collecting the upper layer, And 3) drying the upper layer recovered in the liquid / liquid separation step and recovering the lipid.
  • JP-A-2010-65167 proposes a method for producing high purity plasmalogen-type phospholipids and sphingolipid fractions from animal tissue which can be obtained in large quantities inexpensively.
  • This manufacturing method is A) A process of subjecting an animal tissue containing plasmalogen-type phospholipid and sphingolipid to ethanol extraction to obtain an ethanol extract, B) hydrolyzing a diacyl type glycerophospholipid contained in the ethanol extract obtained in the step (A), C) treating the treated product obtained in the step B) with a water-soluble ketone solvent to recover the insoluble portion, D) A step of solvent-partitioning the insoluble portion obtained in the step C) with a mixed organic solvent of an aliphatic hydrocarbon solvent and a water-soluble ketone solvent and water, and recovering the mixed organic solvent portion.
  • JP 2003-3190 A Japanese Patent Application Publication No. 2003-12520 Japanese Patent Application Publication No. 2004-26803 JP, 2013-53109, A JP, 2007-262024, A (claims) JP, 2010-65167, A (claim)
  • Patent Document 5 The method disclosed in Patent Document 5 is complicated because it requires the use of a wide variety of solvents and the extraction step needs to be performed several times.
  • the method disclosed in Patent Document 6 uses the enzyme phospholipase A1 (PLA1), and this PLA1 utilizes the property that it does not degrade ether type glycerophospholipid. .
  • PPA1 phospholipase A1
  • Such a method comprises the steps of drying the primary extract from the source tissue and then resuspending in an enzyme solution to carry out an enzyme reaction. Therefore, in order to carry out this enzyme reaction more efficiently, it is necessary to prepare an emulsion having good dispersibility.
  • the lipid extract is oil-based while the enzyme solution is water-based, it is not easy to carry out the resuspension.
  • the lipid extract is in a small amount, even if the lipid extract adheres to the container wall, it is possible to carry out resuspension by performing ultrasonic treatment etc. It becomes.
  • the ether type glycerophospholipid excellent in the effect of ameliorating and preventing diseases such as Alzheimer's disease can be produced with high purity and simple operation, and the scale-up also improves the working efficiency and working time, There is a need for a process for producing ether type glycerophospholipids.
  • the present invention has been intensively studied for the purpose of providing a method for efficiently producing a high purity ether type glycerophospholipid by a simple operation.
  • the invention according to claim 1 of this invention is A method for producing an ether type glycerophospholipid comprising treating a biological material or a tissue thereof with a proteolytic enzyme treatment and treating with a lipolytic enzyme simultaneously with or after the treatment with the proteolytic enzyme.
  • the invention described in claim 2 of this invention is in the method for producing ether type glycerophospholipid according to claim 1,
  • the proteolytic enzyme is It is characterized by being exopeptidase, endopeptidase, or a mixture thereof.
  • the invention described in claim 3 of this invention is in the method for producing an ether type glycerophospholipid according to claim 1 or 2,
  • the lipolytic enzyme is It is characterized by being phospholipase A1.
  • the invention described in claim 4 of this invention is in the method for producing an ether type glycerophospholipid according to any one of claims 1 to 3,
  • the biological material or the tissue thereof is It is characterized in that it is of animal origin.
  • the method for producing an ether type glycerophospholipid according to the present invention comprises treating a biological material or a tissue thereof with a proteolytic enzyme to dissolve tissue of the biological material, and treating with a lipolytic enzyme simultaneously with or after the protease treatment. And hydrolyzing the diacyl type glycerophospholipid contained in the tissue of the biological material. Therefore, according to this production method, highly pure ether type glycerophospholipids can be efficiently carried out by simple operation, without performing the primary extraction operation and the operation of removing neutral lipids from the total lipid and the operation of resuspension. You can get it.
  • the method for producing an ether type glycerophospholipid of the present invention can be applied to animal-derived biological materials or tissues thereof.
  • FIG. 2 is an HPLC chart, wherein (a) is an HPLC chart of an ether type glycerophospholipid manufactured based on the method shown in the flow chart of FIG. 1, and (b) is shown in the flow chart of FIG. Fig.
  • FIG. 6 shows an HPLC chart of ether type glycerophospholipid manufactured based on the method. It is a flowchart which shows the other example of the manufacturing method of the ether type glycerophospholipid of this invention, Comprising: The manufacturing method of the scallop string origin ether type glycerophospholipid is shown. It is a flowchart which shows the other example of the manufacturing method of the conventional ether type glycerophospholipid, Comprising: The manufacturing method of the scallop string origin ether type glycerophospholipid is shown.
  • the HPLC chart of the scallop string origin ether type glycerophospholipid obtained by the manufacturing method of the ether type glycerophospholipid of this invention is shown.
  • the HPLC chart of the chicken-derived ether type glycerophospholipid obtained by the manufacturing method of the ether type glycerophospholipid of this invention is shown.
  • the HPLC chart of the yellowtail derived ether type glycerophospholipid obtained by the manufacturing method of the ether type glycerophospholipid of this invention is shown.
  • PE Phosphatidylethanolamine (a kind of diacyl type glycerophospholipid)
  • PC Phosphatidyl choline (a kind of diacyl type glycerophospholipid)
  • CAEP ceramide aminoethyl phosphonic acid
  • SM sphingomyelin (a type of sphingophospholipid)
  • plsPE ethanolamine plasmalogen plsPC: choline plasmalogen PLA1: phospholipase A1 Chol: cholesterol
  • the method for producing an ether type glycerophospholipid of the present invention comprises treating a biological material or a tissue thereof with a proteolytic enzyme and treating with a lipolytic enzyme simultaneously with or after the aforementioned proteolytic enzyme treatment.
  • a lipolytic enzyme for example, PLA1
  • PLA1 is allowed to directly act on biological tissues such as animal tissues, plant tissues or microorganisms as raw materials, or the tissues thereof without performing primary extraction conventionally performed. It becomes possible to hydrolyze mixed diacyl type glycerophospholipids.
  • ether type glycerophospholipid of the present invention high purity ether type glycerophospholipid can be efficiently obtained by a simple operation. Furthermore, since the biological material or the tissue thereof can be in a liquid state in the case of the lipolytic enzyme treatment, it becomes possible to continuously produce a high purity ether type glycerophospholipid.
  • primary extraction refers to an operation for extracting total lipids from the biological material or its tissue before enzyme treatment.
  • ether type glycerophospholipid refers to a glycerophospholipid having a vinyl ether bond (alkenyl bond) or an ether bond (alkyl bond) at position 1 (sn-1) of the glycerol backbone.
  • the general formula of ether type glycerophospholipid is shown.
  • the compound represented by the formula (1) is an alkenyl phospholipid (Plasmalogen)
  • the compound represented by the formula (2) is an alkyl phospholipid.
  • R 1 represents an aliphatic hydrocarbon group.
  • R 1 is usually a C 14-18 aliphatic hydrocarbon group.
  • R 2 is an aliphatic hydrocarbon group, and in some cases, polyunsaturated fatty acids such as arachidonic acid (ARA), docosahesaenoic acid (DHA), and eicosapentaenoic acid (EPA) may be bonded.
  • ARA arachidonic acid
  • DHA docosahesaenoic acid
  • EPA eicosapentaenoic acid
  • X is a nitrogen-containing alcohol group or a polyol group.
  • Examples of the nitrogen-containing alcohol group include hydrogen, a serine group, an ethanolamine group, an N-methylethanolamine group, a dimethylethanolamine group, and a trimethylethanolamine group.
  • Examples of the polyol group include glycerol group, glycerophosphate group, glycerophosphate phosphatidyl group, inositol group, inositol phosphate group, inositol diphosphate group and the like.
  • the biological material to be used is not particularly limited as long as it contains an ether type glycerophospholipid.
  • animals, plants and microorganisms can be mentioned.
  • the biological material it is preferable to select an animal or a tissue thereof because the content of ether type glycerophospholipid is high compared to plant tissues and microorganisms, and it is easy to obtain a large amount inexpensively.
  • the animals include mammals, birds and fish and shellfish.
  • livestock is preferable in terms of both supply stability and safety.
  • mammals such as cows, pigs, horses, goats, sheep, deer, camels, llamas and other poultry, and poultry such as chickens, ducks, turkeys and ostrich.
  • main tissues containing ether type glycerophospholipids include skin, brain, intestine, heart, genitals and the like.
  • those which can be reared, that is, capable of being aquaculture are preferable Yellowtail, red snapper, horse mackerel, horse mackerel, jack mackerel, horse mackerel, horse mackerel, horse mackerel, horse mackerel, horse mackerel, horse mackerel, horse mackerel, horse mackerel, horse mackerel, horse mackerel, horse mackerel, horse mackerel, horse mackerel, horse mackerel, horse mackerel, horse mackerel, horse mackerel, horse mackerel, horse mackerel Crustaceans such as kuruma prawn, black tiger, thai show shrimp, Examples are shellfish such as abalone, clams, scallops and oysters. Among them, shellfish such as abalone, clam, scallop, and oyster are more preferable.
  • scallops because the content of neutral lipids in total lipids is low, the content of phospholipids is high, and the content of ether type glycerophospholipids in phospholipids is also high.
  • main tissues containing ether type glycerophospholipids include internal organs, gonads, muscles and the like.
  • bacteria of the genus Propionibacterium can be used.
  • tissue is bacteria itself.
  • tissue In addition, about the said biological material and its structure
  • tissue processes, such as a boiling and freezing, may be performed from a viewpoint of rot prevention before an enzyme treatment.
  • a treatment such as mincing or crushing may be performed in advance.
  • the biological material or the tissue thereof may be subjected to high-speed homogenizer treatment or homomixer treatment during the enzyme treatment.
  • the proteolytic enzyme treatment is for dissolving (hydrolyzing) the biological material and its tissue.
  • the hydrolysis step and concentration / purification step of the diacyl type glycerophospholipid by a lipolytic enzyme can be simplified as compared with the prior art, and the ether type glycerophospholipid can be extracted efficiently.
  • the biological material as the raw material and its tissue can be made liquid, it is also possible to continuously extract the ether type glycerophospholipid. At that time, extraction can be performed continuously using a single-stage centrifugal extractor, a multistage centrifugal extractor, a mixer-settler or the like.
  • the above-mentioned proteolytic enzyme is not particularly limited as long as it is capable of dissolving (hydrolyzing) the above-mentioned biological material and tissue thereof, and so on. Therefore, it may be selected appropriately according to the biological material to be selected and the tissue thereof.
  • proteolytic enzyme for example, exopeptidase, endopeptidase or a mixture thereof can be used.
  • B) Animal-derived proteases such as pancreatin, trypsin, chymotrypsin and pepsin
  • B) Plant-derived proteases such as papain, bromelain and ficin
  • C) Proteases derived from microorganisms such as Bacillus subtilis, lactic acid bacteria, yeast, mold, Bacillus subtilis, actinomycetes and the like can be mentioned.
  • any protease having an optimum pH in the acidic range (acidic protease), neutral range protease (neutral protease) and alkaline range protease (alkaline protease) can be used, and a mixture of these can be used. It may be When using as a mixture, there is no restriction
  • the said proteolytic enzyme can select suitably according to the quantity of the biological material selected as a raw material. Preferably, it is 10 to 10,000 units, more preferably 100 to 5,000 units, per gram of the biological material.
  • the treatment of the proteolytic enzyme can be carried out according to a conventional method.
  • it can be carried out by a dipping method (eg, cold dipping, digestion), a vacuum dipping method, a pressure dipping method or the like.
  • reaction conditions of the said proteolytic enzyme it can select suitably.
  • an enzyme solution is added, preferably left for 30 minutes or more at a temperature of 50 ° C, more preferably for 1 hour or more at a temperature of 50 ° C. The method of doing is illustrated.
  • an enzyme other than the proteolytic enzyme may be appropriately added so that the proteolytic enzyme easily acts on the selected biological material.
  • enzymes include glycolytic enzymes used to degrade sugars in biological materials.
  • the lipolytic enzyme is for specifically hydrolyzing the sn-1 acyl bond of diacyl type glycerophospholipid.
  • the sn-1 of this ether type glycerophospholipid is an ether bond, so the lipolytic enzyme does not act.
  • the diacyl type glycerophospholipid is decomposed into free fatty acid and lysophospholipid.
  • the degraded free fatty acids and lysophospholipids can be removed utilizing their relatively water solubility.
  • the lipolytic enzyme examples include phospholipase A1 (PLA1) derived from Aspergillus oryzae.
  • PLA1 phospholipase A1
  • Such PLA1 can be purchased from, for example, Mitsubishi Chemical Foods Corporation.
  • the amount of the lipolytic enzyme used is also the same as in the case of the proteolytic enzyme, and may be appropriately selected according to the amount of biological material selected as the raw material. Preferably, it is 10 to 10,000 units, more preferably 100 to 5,000 units, per gram of the biological material. If it is less than 10 units, a sufficient effect tends not to be obtained, and if it exceeds 10,000 units, it is uneconomical.
  • the reaction conditions can be appropriately selected according to the properties such as the optimum temperature and the optimum pH of the enzyme to be used.
  • the temperature is 25 to 60 ° C., pH 4.0 to 8.0, more preferably 30 to 50 ° C., pH 4.5 to 7.0, optionally with stirring, preferably 1 to 24 hours, more preferably
  • the reaction is preferably carried out for 1 to 5 hours, more preferably 1 to 2 hours.
  • the enzyme reaction by the lipolytic enzyme and the proteolytic enzyme can be carried out in a buffer in order to carry out the enzyme reaction under the optimum pH conditions, but such a buffer is appropriately selected depending on the lipolytic enzyme and the proteolytic enzyme used. It can be selected. For example, 250 mM citric acid-HCl buffer (pH 5.2) can be used. In this case, the buffer is added to the biological material, and then the proteolytic enzyme and the lipolytic enzyme (PLA1) are added thereto, or the proteolytic enzyme and the buffer are added to the biological material and dissolved. From this, the above-mentioned lipolytic enzyme (PLA1) may be added.
  • the amount of the buffer used is not particularly limited as long as the enzyme reaction can proceed. Preferably, it is 0.1 to 10 mL, more preferably 0.5 to 5 mL, per 1 g of biological material as a raw material.
  • the enzyme reaction may be stopped by cooling. Furthermore, the enzyme may be inactivated.
  • the above-mentioned inactivation treatment can be carried out by raising the temperature (for example, to about 70 ° C.) according to the characteristics of the enzyme to be used after the hydrolysis reaction.
  • a treatment liquid in which the diacyl type glycerophospholipid is decomposed can be obtained.
  • the enzyme buffer and the enzyme protein can be removed.
  • the production method of the present invention may further include a step of purifying ether type glycerophospholipid.
  • the purification step it is preferable to include the purification step, since it is possible to obtain a purified and concentrated ether type glycerophospholipid having a better effect.
  • the treatment solution in which the diacyl type glycerophospholipid has been degraded obtained after the above-mentioned lipolytic enzyme treatment can be further subjected to a purification step.
  • the purification can be performed according to known methods.
  • an ether type glycerophospholipid is soluble in hexane but hardly soluble in a water-soluble ketone solvent such as acetone, so that these solvents and water are appropriately combined and distributed, and water or an aqueous acetic acid solution, etc.
  • Lysophospholipids can be removed and ether type glycerophospholipids can be purified by solution partitioning (solvent distribution method) with an aqueous acidic solution of That is, neutral lipids other than phospholipids can be removed by a water-soluble ketone solvent such as acetone, and can be separated into ether type glycerophospholipids and lysophospholipids by aqueous solution distribution.
  • hexane / 2-propanol mixed solution 3: 2
  • a 2 to 10-fold solution of hexane / 2-propanol mixed solution 3: 2
  • the treatment solution hydrolysis treatment solution
  • the mixture is transferred to a separatory funnel
  • water By adding water, dividing into two layers and collecting the upper layer (hexane layer), it is possible to remove the lipolysis products (free fatty acid, lysophospholipid), the enzyme protein and the enzyme buffer.
  • neutral lipids other than phospholipids may be further removed by a 5 to 50-fold solution with a water-soluble ketone solvent such as acetone.
  • the ether type glycerophospholipid obtained by the production method of the present invention mainly comprises ethanolamine phospholipid and choline phospholipid.
  • the said ether type glycerophospholipid can be analyzed and confirmed by high performance liquid chromatography (HPLC).
  • Ether-type glycerophospholipids obtained by the method for producing such a composition are extremely useful for the treatment and amelioration of Alzheimer's disease, Parkinson's disease, depression, neurological diseases such as schizophrenia, metabolic syndromes such as diabetes, various infections and immune disorders. It is valid.
  • the ether type glycerophospholipid can be obtained with high purity, particularly 50% or more.
  • the ether type glycerophospholipid can be used as a raw material of food and drink or a raw material of medicine. Such food and drink and medicine may be produced according to known methods.
  • ether type glycerophospholipid various food and drink forms known as described above or developed in the future can be suitably adopted.
  • the form of the functional food or the food for specified health use can be adopted similarly.
  • a pharmaceutically acceptable base, carrier, additive for example, an excipient, a binder, a disintegration, etc.
  • a pharmaceutically acceptable base, carrier, additive for example, an excipient, a binder, a disintegration, etc.
  • Agents, lubricants, solvents, sweeteners, coloring agents, flavoring agents, flavoring agents, surfactants, moisturizers, preservatives, pH adjusters, thickening agents, etc. can be blended.
  • Such base materials, carriers, additives and the like are specifically described, for example, in the Pharmaceutical Additives Dictionary 2000 (Yakuji Nipponsha Co., Ltd.), and for example, those described therein can be used.
  • the form of the preparation is also not particularly limited, and the active ingredient and other ingredients are mixed in a conventional manner, for example, tablets, coated tablets, powders, granules, fine granules, capsules, pills, solutions, suspensions, It can be prepared into a formulation such as an emulsion, a jelly, a chewable, a soft tablet and the like.
  • the ether type glycerophospholipid is conveniently used by being mixed with the product to be used, but it is natural that the ether type glycerophospholipid should be contained in an effective amount to exert the above-mentioned function.
  • an amount such that the amount of ether type glycerophospholipid in the product is preferably in the range of preferably 0.01 to 100 mg, more preferably 0.05 to 50 mg, still more preferably 0.1 to 10 mg per adult day It is preferable to In addition, it may be divided and taken once a day or a plurality of times (preferably 2 to 3 times).
  • Example 1 (Production of scallop-derived ether type glycerophospholipid) (1) Extraction of Scallop String-Derived Ether Type Glycerophospholipid Based on the flow chart shown in FIG. 1, extraction of the scallop string-derived ether type glycerophospholipid was carried out.
  • Fresh scallops of 0.2 kg fresh weight are cut and cut into enzyme solution (1% coclase P (registered trademark; manufactured by Mitsubishi Chemical Foods), 2% PLA 1, 0.1 M citrate buffer, pH 4.5) 50 mL) was crushed using a blender, homogenized, and treated at a temperature of 50 ° C. for 1 hour.
  • 1.0 L of a mixed solution of hexane / 2-propanol (3: 2) was added, and stirred for 10 minutes. Then, suction filtration was performed, the residue was washed with a mixture of 0.2 L of hexane / 2-propanol (3: 2), and the combined filtrate was transferred to a separatory funnel.
  • Comparative Example 1 (Production of scallop-derived ether type glycerophospholipid) (1) Extraction of Scallop String-Derived Ether-Type Glycerophospholipid [0163] Based on the flow chart shown in FIG. 2, extraction of the scallop string-derived ether-type glycerophospholipid was performed.
  • a fresh weight of 0.2 kg of fresh scallop is cut, 1.0 L of a mixed solution of hexane / 2-propanol (3: 2) is added to this, and the mixture is crushed and homogenized using a blender, and then stirred at room temperature 1 I put time. Then, suction filtration was performed, the residue was washed with 0.4 L of hexane / 2-propanol (3: 2) mixture, and the combined filtrate was transferred to a separatory funnel. After adding 1.0 L of sodium sulfate (1 g / 15 mL) to the separation funnel and mixing, it was allowed to stand.
  • the lower layer separated into two layers was discarded, the upper hexane layer was recovered, and the obtained hexane layer was dried with a rotary evaporator to obtain a dried product.
  • the treatment liquid was transferred to a separatory funnel, and 360 mL of a mixed solution of hexane / 2-propanol (3: 2) and 220 mL of water were added and allowed to stand.
  • the lower layer separated into two layers was discarded, and the upper hexane layer was recovered.
  • the obtained hexane layer was dried by a rotary evaporator to obtain a crude extract as a lipid fraction. Furthermore, 20 volumes (v / w) of acetone was added to the obtained crude extract, and after thoroughly stirring, the mixture was allowed to stand still in a freezer at a temperature of -30 ° C for 1 hour or more, and cooled. The precipitate was recovered by centrifugation at 5,000 ⁇ g for 5 minutes, and acetone was completely distilled off under reduced pressure to obtain a purified scallop-derived ether type glycerophospholipid (purified extract).
  • the manufacturing method of the first embodiment does not perform the primary extraction operation conventionally performed, and the number of times of the concentration step is smaller than that of the manufacturing method of the comparative example 1.
  • the lipid obtained in Example 1 contains almost no diacyl type glycerophospholipid, and contains ether type glycerophospholipid in high purity, and is obtained in Comparative Example 1.
  • the time taken to obtain the ether type glycerophospholipid in Example 1 is about 3.5 hours
  • the time taken to obtain the ether type glycerophospholipid in Comparative Example 1 is It was about 7 hours.
  • the production process can be shortened compared to the prior art, and thus ether type glycerophospholipid can be obtained in a short time and in a simple operation. Furthermore, while the yield of the ether type glycerophospholipid obtained in Example 1 is 6.6 g / kg, the yield of the ether type glycerophospholipid obtained in Comparative Example 1 is 5.2 g / kg. there were. Therefore, it was also shown that according to the production method of the present invention, ether type glycerophospholipid can be obtained in high yield as compared with the conventional. Therefore, according to the production method of the present invention, ether type glycerophospholipid can be efficiently obtained by a simple operation.
  • Example 2 (Production of scallop-derived ether type glycerophospholipid) (1) Extraction of Scallop String-Derived Ether-Type Glycerophospholipid [0163] Based on the flow chart shown in FIG. 4, extraction of the scallop-derived ether-type glycerophospholipid was performed.
  • the manufacturing method of the second embodiment does not perform the primary extraction operation conventionally performed, and the number of times of the concentration step is smaller than that of the manufacturing method of the comparative example 2.
  • the lipid obtained in Example 2 contains almost no diacyl type glycerophospholipid, and contains ether type glycerophospholipid in high purity, and is obtained in Comparative Example 2.
  • Example 3 (Production of scallop-derived ether type glycerophospholipid) (1) Extraction of Scallop String-Derived Ether-Type Glycerophospholipid Scallop string-derived ether-type glycerophospholipid was extracted based on the flow chart shown in FIG.
  • Fresh weight 1.0 kg of raw scallop string was placed in boiling water and heat treated for 2 minutes to obtain 0.345 kg of boiled scallop string.
  • an enzyme solution (1.5% coclase ⁇ P (registered trademark) (manufactured by Mitsubishi Chemical Foods), 1.5% PLA 1, 0.25 M citrate buffer, pH 5.2)
  • the solution obtained by adding 690 mL of the above solution was sealed in a plastic bag (No. 1-8344-13: manufactured by As One Corporation) for a vacuum pack and subjected to enzyme treatment at a temperature of 50 ° C. for 5 hours.
  • the treated product obtained was a liquid with almost no solid content.
  • Example 4 (Production of scallop-derived ether type glycerophospholipid) (1) Extraction of ether type glycerophospholipid derived from scallop string A fresh scallop string of 20 kg fresh weight was placed in boiling water and heat-treated for 2 minutes to obtain 4.89 kg of boil scallop string. The obtained boiled scallop strings are divided into about 1 kg portions and put in a poly pack, and each of the enzyme solutions (1.5% coclase P (registered trademark) (made by Mitsubishi Chemical Foods), 1.5% PLA 1, 0.25 M) Citrate buffer solution, pH 5.2) was added equally to make a total of 9.78 L, and sealed. Enzyme treatment was carried out at a temperature of 50 ° C. for 5 hours. The treated product obtained was a liquid with almost no solid content.
  • the treatment solution was stored frozen at a temperature of -30 ° C for 24 hours or more.
  • the treated solution was thawed, and 35 L of a mixed solution of hexane / 2-propanol (3: 2) was added to it, and stirred for 1 hour. Thereafter, the mixture was transferred to a separatory funnel, and 20 L of sodium sulfate (1 g / 15 mL) was added and mixed, and then stirred for 5 minutes and allowed to stand.
  • the lower layer separated into two layers was discarded, and about 21 L of the upper hexane layer was recovered.
  • the obtained hexane layer was dried by a rotary evaporator to obtain about 100 g of a crude extract as a lipid fraction.
  • Example 5 Production of chicken-derived ether type glycerophospholipid
  • an enzyme solution (1.5% coclase P (registered trademark; manufactured by Mitsubishi Chemical Foods Co., Ltd.), 0.25 L of 1.5% PLA 1, 0.25 M citrate buffer, pH 5.2) was added, and after crushing and homogenization using a blender, treatment was performed at a temperature of 50 ° C. for 5 hours.
  • To the treating solution was added 1.2 L of a mixed solution of hexane / 2-propanol (3: 2), and stirred for 15 minutes.
  • the treatment solution was transferred to a separatory funnel, and 0.8 L of sodium sulfate (1 g / 15 mL) was added and mixed, followed by stirring for 5 minutes. After standing at room temperature, the lower layer separated into two layers was discarded, and the upper hexane layer was recovered. The obtained hexane layer was dried by a rotary evaporator to obtain 31.7 g of a crude extract as a lipid fraction. To the obtained crude extract was added 0.15 L of acetone, and after thorough stirring, the mixture was allowed to stand still for 1 hour or more at a temperature of -30 ° C and cooled. The precipitate was collected by centrifugation at 4,000 ⁇ g for 10 minutes. This operation was repeated twice to obtain 0.1 g of a chicken-derived ether type glycerophospholipid (purified extract) purified as an acetone-insoluble fraction.
  • the lipid (product of the present invention) obtained in Example 5 contained almost no diacyl type glycerophospholipid and contained an ether type glycerophospholipid in high purity. Therefore, according to the production method of the present invention, even when chicken meat is selected as the biological material, ether type glycerophospholipid can be efficiently obtained by a simple operation.
  • the lipid obtained in Example 5 includes a lipid presumed to be a sphingolipid ("SM" in FIG. 10), separation of such sphingolipid and ether type glycerophospholipid is also included.
  • SM sphingolipid
  • Example 6 (Production of yellowtail-derived ether type glycerophospholipid) (1) Extraction of yellowtail-derived ether type glycerophospholipid After cutting 0.20 kg of fresh fresh weight of yellowtail, an enzyme solution (1.5% coclase P (registered trademark; manufactured by Mitsubishi Chemical Foods Co., Ltd.), 1.5) 0.20 L of 1% PLA 1, 0.25 M citrate buffer, pH 5.2) was added, and treatment was carried out at a temperature of 50 ° C. for 5 hours. To the treating solution was added 1.2 L of a mixed solution of hexane / 2-propanol (3: 2), and stirred for 15 minutes.
  • an enzyme solution (1.5% coclase P (registered trademark; manufactured by Mitsubishi Chemical Foods Co., Ltd.), 1.5) 0.20 L of 1% PLA 1, 0.25 M citrate buffer, pH 5.2
  • To the treating solution was added 1.2 L of a mixed solution of hexane / 2-propanol (3: 2), and stirred for 15 minutes.
  • the treatment solution was transferred to a separatory funnel, and 0.8 L of sodium sulfate (1 g / 15 mL) was added and mixed, followed by stirring for 5 minutes. After standing at room temperature, the lower layer separated into two layers was discarded, and the upper hexane layer was recovered. The obtained hexane layer was dried by a rotary evaporator to obtain 50.3 g of a crude extract as a lipid fraction. To the obtained crude extract was added 0.4 L of acetone, and after thorough stirring, the mixture was allowed to stand still for 1 hour or more at a temperature of -30.degree. C. and cooled. The precipitate was collected by centrifugation at 4,000 ⁇ g for 10 minutes. This operation was repeated twice to obtain 0.19 g of yellowtail-derived ether type glycerophospholipid (purified extract) purified as an acetone-insoluble fraction.
  • an ether type glycerophospholipid effective for amelioration and prevention of Alzheimer's disease and the like can be efficiently produced with high purity and simple operation, and therefore, is widely used in the pharmaceutical industry. .

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Abstract

Le problème à la base de l'invention concerne un procédé de production d'un glycérophospholipide de type éther de haute pureté, à une efficacité élevée, par un procédé simple. La solution porte sur un procédé de production, qui consiste à traiter une matière biologique ou un tissu correspondant avec une protéase pour lyser le tissu de la matière biologique et, pendant ou après le traitement avec la protéase, à traiter le lysat avec une enzyme lipolytique pour hydrolyser un glycérophospholipide de type diacyle contenu dans le tissu de la matière biologique. Il devient ainsi possible de produire un glycérophospholipide de type éther de haute pureté à une efficacité élevée par un procédé simple, qui ne nécessite pas d'opération d'extraction primaire, c'est-à-dire l'extraction de la totalité des lipides avant un traitement avec un enzyme, ni une opération pour éliminer un lipide neutre de tous les lipides ou une opération de remise en suspension.
PCT/JP2015/086032 2015-12-24 2015-12-24 Procédé de production glycérophospholipide de type éther Ceased WO2017109897A1 (fr)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020213132A1 (fr) * 2019-04-18 2020-10-22 株式会社レオロジー機能食品研究所 Procédé de production d'un matériau fonctionnel contenant du plasmalogène
WO2024084851A1 (fr) * 2022-10-17 2024-04-25 株式会社 レオロジー機能食品研究所 Milieu de culture favorisant la production de plasmalogène, procédé de production de composition contenant du plasmalogène et procédé de production de plasmalogène

Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS49118866A (fr) * 1973-03-26 1974-11-13
JPS6431895A (en) * 1987-07-27 1989-02-02 Asahi Denka Kogyo Kk Production of fish oil
JPS6431896A (en) * 1987-07-10 1989-02-02 Novo Industri As Extraction of oils and fats from animal fatty tissue using protease
JPH04252154A (ja) * 1991-01-25 1992-09-08 Fine Kagaku Kenkyusho:Kk 卵黄油の製造法
JPH11123052A (ja) * 1997-10-24 1999-05-11 Asahi Chem Ind Co Ltd 魚介類由来の有用物質を含む組成物およびその有用物質の製造方法
JP2009227601A (ja) * 2008-03-21 2009-10-08 Adeka Corp 血圧降下剤
JP2010065167A (ja) * 2008-09-11 2010-03-25 Marudai Food Co Ltd プラズマローゲン型リン脂質及びスフィンゴ脂質の製造方法
JP2013053109A (ja) * 2011-09-05 2013-03-21 Teikyo Univ リン脂質結合型ｄｈａ増加剤
JP2014509339A (ja) * 2011-02-11 2014-04-17 テドク・エフアールディー・カンパニー・リミテッド オキアミ油の製造方法およびこの方法で製造されたオキアミ油

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP5357362B2 (ja) * 2010-12-14 2013-12-04 大▲連▼工▲業▼大学 ナンキョクオキアミからオキアミオイル又はオキアミ濃縮物、オキアミミールを製造する方法
CN103087093A (zh) * 2011-11-02 2013-05-08 江南大学 一种猪肺磷脂的制备方法
CN104357206A (zh) * 2014-11-18 2015-02-18 董寰 一种水酶法制备富含磷脂的南极磷虾油的方法

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS49118866A (fr) * 1973-03-26 1974-11-13
JPS6431896A (en) * 1987-07-10 1989-02-02 Novo Industri As Extraction of oils and fats from animal fatty tissue using protease
JPS6431895A (en) * 1987-07-27 1989-02-02 Asahi Denka Kogyo Kk Production of fish oil
JPH04252154A (ja) * 1991-01-25 1992-09-08 Fine Kagaku Kenkyusho:Kk 卵黄油の製造法
JPH11123052A (ja) * 1997-10-24 1999-05-11 Asahi Chem Ind Co Ltd 魚介類由来の有用物質を含む組成物およびその有用物質の製造方法
JP2009227601A (ja) * 2008-03-21 2009-10-08 Adeka Corp 血圧降下剤
JP2010065167A (ja) * 2008-09-11 2010-03-25 Marudai Food Co Ltd プラズマローゲン型リン脂質及びスフィンゴ脂質の製造方法
JP2014509339A (ja) * 2011-02-11 2014-04-17 テドク・エフアールディー・カンパニー・リミテッド オキアミ油の製造方法およびこの方法で製造されたオキアミ油
JP2013053109A (ja) * 2011-09-05 2013-03-21 Teikyo Univ リン脂質結合型ｄｈａ増加剤

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020213132A1 (fr) * 2019-04-18 2020-10-22 株式会社レオロジー機能食品研究所 Procédé de production d'un matériau fonctionnel contenant du plasmalogène
JP2020176086A (ja) * 2019-04-18 2020-10-29 株式会社 レオロジー機能食品研究所 脳神経炎症抑制組成物の製造方法
CN112105739A (zh) * 2019-04-18 2020-12-18 株式会社流变机能食品研究所 包含缩醛磷脂的功能性素材的制造方法
US11491170B2 (en) 2019-04-18 2022-11-08 Institute of Rheological Functions of Food Method for manufacturing functional material including plasmalogen
EP3957737A4 (fr) * 2019-04-18 2022-12-14 Institute Of Rheological Functions Of Food Procédé de production d'un matériau fonctionnel contenant du plasmalogène
WO2024084851A1 (fr) * 2022-10-17 2024-04-25 株式会社 レオロジー機能食品研究所 Milieu de culture favorisant la production de plasmalogène, procédé de production de composition contenant du plasmalogène et procédé de production de plasmalogène

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