WO2013146529A1 - Procédé de production de graisse ou d'huile interestérifiée de manière aléatoire - Google Patents

Procédé de production de graisse ou d'huile interestérifiée de manière aléatoire Download PDF

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Publication number
WO2013146529A1
WO2013146529A1 PCT/JP2013/058074 JP2013058074W WO2013146529A1 WO 2013146529 A1 WO2013146529 A1 WO 2013146529A1 JP 2013058074 W JP2013058074 W JP 2013058074W WO 2013146529 A1 WO2013146529 A1 WO 2013146529A1
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WO
WIPO (PCT)
Prior art keywords
lipase
oil
fats
oils
random
Prior art date
Application number
PCT/JP2013/058074
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English (en)
Japanese (ja)
Inventor
洋介 矢島
晴康 木田
真晴 加藤
Original Assignee
不二製油株式会社
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 不二製油株式会社 filed Critical 不二製油株式会社
Publication of WO2013146529A1 publication Critical patent/WO2013146529A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/18—Carboxylic ester hydrolases (3.1.1)
    • C12N9/20—Triglyceride splitting, e.g. by means of lipase
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23D—EDIBLE OILS OR FATS, e.g. MARGARINES, SHORTENINGS, COOKING OILS
    • A23D9/00—Other edible oils or fats, e.g. shortenings, cooking oils
    • A23D9/02—Other edible oils or fats, e.g. shortenings, cooking oils characterised by the production or working-up
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11C—FATTY ACIDS FROM FATS, OILS OR WAXES; CANDLES; FATS, OILS OR FATTY ACIDS BY CHEMICAL MODIFICATION OF FATS, OILS, OR FATTY ACIDS OBTAINED THEREFROM
    • C11C3/00—Fats, oils, or fatty acids by chemical modification of fats, oils, or fatty acids obtained therefrom
    • 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
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
    • C12P7/6436—Fatty acid esters

Definitions

  • the present invention relates to a novel method for producing fats and oils. More specifically, the present invention relates to a method for producing a random transesterified oil using a lipase obtained by culturing bacteria belonging to Burkholderia plantarii.
  • the transesterification reaction of fats and oils is an effective method for improving the physical properties of fats and oils, and mainly comprises chemical methods and enzymatic methods.
  • the chemical method includes a random transesterification reaction using a chemical catalyst, but this method requires washing with water to remove the catalyst after the reaction is completed, and the operation is very troublesome.
  • the enzymatic method does not require washing with water, it is necessary to perform the reaction at a temperature equal to or higher than the melting point of the fats and oils used. Therefore, when the heat resistance of the lipase is low, the lipase is easily deactivated and efficient. There is a problem that can not react properly.
  • Non-Patent Document 1 discloses a random transesterification reaction using a Candida lipase
  • Patent Document 1 discloses a fat and oil random ester using a lipase derived from Burkholderia gladioli. Exchange reactions are each disclosed.
  • Patent Document 2 reports an example of an enantioselective ring-opening method from oxetane-2-one using a lipase derived from Burkholderia plantarii. Using this lipase, a random triglyceride is reported. No examples of transesterification have been reported.
  • Non-Patent Document 1 is a method in which a large amount of enzyme of 10% or more is added to the fats and oils of the substrate and the reaction is carried out at 40 ° C. for 66 hours, and industrialization is difficult.
  • the lipase used for random transesterification in Patent Document 1 also has room for improvement in heat resistance. Under such circumstances, a lipase with high heat resistance, which can be transesterified with oil and fat at a high temperature, which cannot be solved by existing lipases, has been desired.
  • the present inventors searched for a fungus that produces a novel lipase that exhibits an efficient random transesterification reaction of fats and oils, a lipase derived from Burkholderia plantarii produces a lipase with high heat resistance, and uses this lipase
  • the present invention was completed by successfully producing an efficient random transesterified oil and fat. That is, the present invention (1) A method for producing random transesterified fats and oils using lipase produced by Burkholderia plantarii. (2) The method for producing a random transesterified oil or fat according to claim 1, wherein the oil or fat is a triacylglyceride.
  • a lipase derived from Burkholderia plantarii is allowed to act on fats and oils, whereby it is possible to obtain fats and oils subjected to random transesterification at a high temperature and for a long period of time.
  • the lipase used in the present invention is derived from Burkholderia plantarii. Specifically, a lipase obtained by culturing bacteria obtained by isolation from soil, distribution from an organization, etc., in a normal medium can be used. This lipase is preferably used by being immobilized on a carrier such as diatomaceous earth, silica, ceramic, activated carbon, or ion exchange resin. Such an immobilized lipase can be prepared by bringing the lipase solution into contact with the carrier by infiltration or spraying and then drying by ventilation, reduced pressure, heating or the like.
  • the fats and oils obtained in the present invention are glycerin fatty acid esters which are triacyl glycerides, diacyl glycerides or monoacyl glycerides, and the most suitable modification is to obtain triacyl glycerides.
  • As the substrate used in the present invention various glycerin fatty acid esters can be used.
  • Vegetable fats such as monkey fat
  • animal and vegetable fats such as fish oil, lard, beef tallow, milk fat, and their fractionated fats, hardened oils, transesterified oils, and MCT, trilaurin, triolein, tris
  • examples include triacylglycerides containing synthetic fats and oils such as palmitic acid and tristearin.
  • diacylglycerol esters and monoacylglycerol esters can also be used.
  • long chain saturated fatty acids such as stearic acid, palmitic acid, behenic acid, myristic acid, lauric acid, long chain unsaturated fatty acids such as oleic acid, linoleic acid, linolenic acid, palmitoleic acid, EPA, DHA, and capric acid , Caprylic acid, caproic acid, butyric acid, acetic acid and other medium chain or short chain fatty acids, and these short chain alcohol esters can be used in combination with the glycerin fatty acid ester. (Transesterification reaction)
  • transesterification reaction between the fatty glycerin esters as substrates preferably between the triacylglycerides, or between the fatty glycerin ester, preferably the triacylglyceride, and the fatty acid or fatty acid ester is carried out by transesterification of the present invention. Corresponds to the reaction.
  • Random transesterification reaction using the lipase of the present invention refers to a primary hydroxyl group of glycerin between fatty acids bound to glycerin, or between fatty acids bound to glycerin and free fatty acids or short-chain alcohol esters of fatty acids, By exchanging the bonding position regardless of the secondary hydroxyl group, the reaction is performed by transesterification so that the fatty acid composition at each position becomes the same, thereby modifying the fats and oils. “Randomized” means that the fatty acid composition at each position is the same.
  • the reaction using the lipase may be a batch type or a continuous type, but it is preferable to reduce the substrate moisture to 1% by weight or less because production of a hydrolyzate is effectively suppressed.
  • the amount of enzyme used for fats and oils is preferably from 50 ppm to 10,000 ppm, more preferably from 200 ppm to 2,000 ppm, based on the raw material fats and oils.
  • the reaction can be carried out in the absence of a solvent or in various solvents, but the reaction in the absence of a solvent or in a nonpolar solvent such as hexane is preferred, and the reaction in the absence of solvent is most preferred.
  • the reaction temperature is preferably 30 to 100 ° C, more preferably 40 to 80 ° C, and most preferably 60 to 75 ° C. Since the lipase of the present invention has high heat resistance, it is possible to carry out the reaction for a long period of time with little decrease in activity due to heat in both batch and continuous methods.
  • Each activity measurement of the hydrolysis activity and transesterification activity used in the Examples was performed by the following methods.
  • a) Hydrolysis activity This was carried out by partially modifying the fat digestion test method described in the Japanese Pharmacopoeia. The enzyme was allowed to react with an olive oil substrate (phosphate buffer, pH 7.0) at 37 ° C. for 30 minutes, and then the amount of enzyme that liberates 1 ⁇ mol of fatty acid per minute was 10 Units. When measuring the temperature dependence of activity, the reaction temperature was changed.
  • Random transesterification activity 2g of immobilized lipase was added to 100g of the substrate containing 6: 4 palm mid-melting fraction and palm low-melting fraction, and the mixture was allowed to act at 60 ° C. Asked.
  • (Randomization rate) ((tripalmitin content of reaction product) ⁇ (tripalmitin content of raw oil) / ((tripalmitin content of reaction equilibrium composition) ⁇ (tripalmitin content of raw oil)) ⁇ 100
  • the lipase of the present invention which is Example 1 of the lipase QLM of Comparative Example 1, has high reactivity at high temperatures and high heat resistance.
  • the lipase 0.04 wt% aqueous solution obtained by cultivation of Burkholderia plantarii used in Example 1 was stored at 5 ° C, 40 ° C, and 60 ° C for 24 hours, and then the hydrolysis activity was measured to determine the heat resistance of the lipase. The results are shown in Table 2 (Example 2). As a comparison, the same measurement was performed with lipase QLM, which is a lipase derived from Burkholderia gladioli (Comparative Example 2).
  • the lipase of the present invention was found to have high heat resistance with little inactivation at high temperatures.
  • Example 4 2 g of immobilized lipase derived from Burkholderia plantarii used in Example 3 was added to 100 g of palm super olein (IV65) and reacted at 60 ° C. for 24 hours (Example 4).
  • a 1,3-selective lipase, Neurase F3GR from Rhizopus niveus, Amano Enzyme was added to the oil and reacted in the same manner (Comparative Example 3).
  • a random transesterified oil produced using a chemical catalyst (Comparative Example 4) was prepared.
  • Table 3 shows the solid fat content (SFC) and the tripalmitin (PPP) content in the triacylglyceride (TG) composition for the raw material oil and the total of 4 points of Example 4 and Comparative Examples 3 to 4.
  • the lipase of the present invention catalyzes a strong random transesterification reaction, it can be used for oil and fat modification. These transesterified oils are useful as safe transesterified fats because they do not use chemical catalysts.

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Health & Medical Sciences (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Chemical & Material Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Polymers & Plastics (AREA)
  • Food Science & Technology (AREA)
  • Biomedical Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

L'invention concerne une lipase hautement résistante à la chaleur, qui permet une interestérification aléatoire à haute température d'une graisse ou d'une huile. D'ailleurs une interestérification aléatoire à haute température d'une graisse ou d'une huile ne peut être réalisée avec une lipase classique. Une lipase provenant de Burkholderia plantarii peut maintenir l'activité même à haute température, ce permet d'utiliser la lipase pour produire efficacement une huile ou une graisse interestérifiée de manière aléatoire. La graisse ou l'huile à produire par ce procédé est, de préférence, un triacylglycéride.
PCT/JP2013/058074 2012-03-28 2013-03-21 Procédé de production de graisse ou d'huile interestérifiée de manière aléatoire WO2013146529A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2012-073168 2012-03-28
JP2012073168 2012-03-28

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WO2013146529A1 true WO2013146529A1 (fr) 2013-10-03

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021020458A1 (fr) * 2019-08-01 2021-02-04 天野エンザイム株式会社 Nouvelle lipase et son utilisation

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0468102A1 (fr) * 1989-05-01 1992-01-29 Solvay Enzymes, Inc. Formulations détergentes contenant une lipase alcaline
JPH08332093A (ja) * 1995-06-07 1996-12-17 Asahi Denka Kogyo Kk エステル交換油脂の製造方法
JPH11253157A (ja) * 1998-03-09 1999-09-21 Meito Sangyo Co Ltd 新規リパーゼ生産能をもつ細菌、リパーゼ、その製造方法、およびその使用
JP2009540848A (ja) * 2006-06-27 2009-11-26 ビーエーエスエフ ソシエタス・ヨーロピア エステラーゼ活性を有するタンパク質
WO2010085975A1 (fr) * 2009-01-29 2010-08-05 Nordmark Arzneimittel Gmbh & Co. Kg Préparation pharmaceutique contenant des lipases d'origine bactérienne

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0468102A1 (fr) * 1989-05-01 1992-01-29 Solvay Enzymes, Inc. Formulations détergentes contenant une lipase alcaline
JPH08332093A (ja) * 1995-06-07 1996-12-17 Asahi Denka Kogyo Kk エステル交換油脂の製造方法
JPH11253157A (ja) * 1998-03-09 1999-09-21 Meito Sangyo Co Ltd 新規リパーゼ生産能をもつ細菌、リパーゼ、その製造方法、およびその使用
JP2009540848A (ja) * 2006-06-27 2009-11-26 ビーエーエスエフ ソシエタス・ヨーロピア エステラーゼ活性を有するタンパク質
WO2010085975A1 (fr) * 2009-01-29 2010-08-05 Nordmark Arzneimittel Gmbh & Co. Kg Préparation pharmaceutique contenant des lipases d'origine bactérienne

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021020458A1 (fr) * 2019-08-01 2021-02-04 天野エンザイム株式会社 Nouvelle lipase et son utilisation
JP6846577B1 (ja) * 2019-08-01 2021-03-24 天野エンザイム株式会社 新規リパーゼ及びその用途
US11718837B2 (en) 2019-08-01 2023-08-08 Amano Enzyme Inc. Lipase and uses of the same

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