The present invention discloses a microbial polyunsaturated fatty acid(PUFA)-containing oil with a high triglyceride content and a high oxidative stability. In addition, a method is described for the recovery of such oil from a microbial biomass derived from a pasteurised fermentation broth, wherein the microbial biomass is subjected to extrusion to form granular particles, dried and the oil then extracted from the dried granules using an appropriate solvent.

Preparation of microbial polyunsaturated fatty acid containing oil from pasteurised biomass

A method for degumming an oil composition comprises the steps of (a) providing an oil composition containing a quantity of phospholipids, (b) contacting said oil composition simultaneously with one or more phospholipase A enzymes and one or more phospholipase C enzymes, under conditions sufficient for the enzymes to react with the phospholipids to create phospholipid reaction products, and (c) separating the phospholipids reaction products from the oil composition, the remaining oil composition after the separation being a degummed oil composition, whereby during step (b) the reaction of said one or more phospholipase A enzymes proceeds at a faster rate than it would in the absence of said one or more phospholipase C enzymes. In a preferred embodiment the reaction of step (b) continues for a duration of less than about one hour.

Enzymatic Degumming Utilizing a Mixture of PLA and PLC Phospholipases

A process of enzymatic degumming edible oils, comprising treating the edible oil with a lipid acyltransferase so as to transfer an acyl group from a major part of the phospholipid to one or more acyl acceptors, wherein the acyl acceptor may be any compound comprising a hydroxy group. In one embodiment preferably the acyl acceptor is water and in another embodiment preferably the acyl acceptor is one or more sterols and/or stanols. When the acyl acceptor is a stanol and/or sterol, one or more sterol esters and/or stanol esters are produced. The lipid acyltransferase for use in the process of the present invention may comprises one or more of the following amino acid sequences: SEQ ID No. 1, SEQ ID No. 3, SEQ ID No. 4, SEQ ID No. 5, SEQ ID No. 6, SEQ ID No. 7, SEQ ID No. 8, SEQ ID No. 9, SEQ ID No. 10, SEQ ID No. 11, SEQ ID No. 12, SEQ ID No. 13, SEQ ID No. 14, or SEQ ID No. 15, SEQ ID No. 16, SEQ ID No. 17, SEQ ID No. 18, SEQ ID No. 36, SEQ ID No. 38, SEQ ID No. 40, SEQ ID No. 41, SEQ ID No 45, SEQ ID No. 47, SEQ ID No. 50 or an amino acid sequence which has 75% or more identity thereto. A novel lipid acyltransferase comprising the amino acid sequence shown as SEQ ID No. 16 is also taught.

Enzymatic oil-degumming method

Edible fats containing arachidonic acid obtained from microorganisms belonging to the subgenus Mortierella of the genus Mortierella and being capable of producing arachidonic acid, containing little unsaponified matters and, above all, the smallest possible amount of cyclopropane sterols which have never been eaten, and being suitable for the production of foods, in particular, modified milks for infants. The fats contain not more than 0.8 % by weight, preferably not more than 0.6 % by weight of unsaponified matters and 20 % by weight or more of arachidonic acid originating in microorganisms. Further, these fats contain not more than 0.3 % by weight, preferably not more than 0.15 % by weight of 24,25-methylenecholest-5-en-3β-ol. The microorganisms are those belonging to the subgenus Mortierella of the genus Mortierella and being capable of producing arachidonic acid. These microorganisms belong to the species alpina of the genus Mortierella. The foods include modified milks for premature infants, modified milks for infants, foods for infants, and foods for pregnant women and nursing mothers containing the above-mentioned edible fats.

Edible fats containing arachidonic acid and foods containing the same

A process of water degumming an edible oil (preferably a crude edible oil) comprising the steps of: a) admixing approximately 0.1-5% w/w water with an edible oil (preferably a crude edible oil) and a lipid acyltransferase, b) agitating the admixture for between about 10 minutes and 180 minutes at about 45 to about 900C, and c) separating the oil phase and the gum phase. Preferably said lipid acyltranferase is a polypeptide having lipid acyltransferase activity which polypeptide is obtained by expression of the nucleotide sequence shown as SEQ ID No. 49 or a nucleotide sequence which as has 70% or more identity therewith; and/or is obtained by expression of a nucleic acid which hybridises under medium stringency conditions to a nucleic probe comprising the nucleotide sequence shown as SEQ ID No. 49; and/or is a polypeptide having lipid acyltranferase activity which polypeptide comprises the amino acid sequence shown as SEQ ID No. 68 or an amino acid sequence which as has 70% or more identity therewith. In one embodiment the lipid acyltransferase is preferably used in combination with a phospholipase C enzyme. A process for modifying the gum phase of a degummed oil using a lipid acyltransferase is also taught herein.

Process for edible oil refining using a lipid acyltransferase

PROBLEM TO BE SOLVED: To provide a degumming method whereby phospholipids can be hydrolyzed more efficiently with phospholipase even at high temps. (e.g. 70 deg.C) or higher); and phospholipase can be recovered and reused. SOLUTION: In a method for removing a gummy substance from an oil or fat by hydrolyzing phospholipids in the oil or fat with phospholipase A1 and/or A2 and removing the resultant hydrolyzate, the hydrolysis is conducted by using phospholipase immobilized on a cation exchanger comprising a hydrophobic support and cation exchange groups. Thus, the degumming can be carried out efficiently even at such a high temp. that the oil or fat loses its viscosity. Since an immobilized enzyme can be recovered and recycled, the degumming is done economically.

Degumming method with phospholipase

PURPOSE:To provide a high-purity arachidonic acid triglyceride and its production process. CONSTITUTION:The objective high-purity arachidonic acid triglyceride is a purified reaction product obtained by reacting arachidonic acid with glycerol in the presence of an immobilized lipase and has an arachidonic acid triglyceride concentration of >=99wt.%. The arachidonic acid triglyceride is produced by mixing arachidonic acid with glycerol at a stoichiometric ratio or thereabout and contacting the obtained substrate with an immobilized lipase to effect the reaction while removing water produced by the reaction as by-product to keep the water-concentration in the reaction liquid to <=1,000ppm.

High-purity arachidonic acid triglyceride and its production

PURPOSE:To obtain the subject oil and fat containing highly unsaturated fatty acid at a high concentration and useful as a preventive for circulatory diseases, functional food, etc., from an inexpensive raw material in high efficiency by a specific method using an oil and fat containing highly unsaturated fatty acid as a raw material. CONSTITUTION:An oil and fat containing highly unsaturated fatty acid (preferably an oil and fat rich in OMEGA-3 highly unsaturated fatty acid triglyceride such as fish oil) or an alcohol ester of a highly unsaturated fatty acid is hydrolyzed with an immobilized lipase (preferably produced by immobilizing a lipase of a microorganism of genus Pseudomonas to an anion exchange resin). The highly unsaturated fatty acid in the hydrolyzed product is concentrated e.g. by a low-temperature fractional crystallization. The concentrated highly unsaturated fatty acid is converted to triglyceride by reacting with glycerol in the presence of an immobilized lipase (preferably produced by immobilizing a lipase of microorganism of genus Candida to an acrylic resin) to obtain the objective oil and fat.

Production of oil and fat containing concentrated highly unsaturated fatty acid

PROBLEM TO BE SOLVED: To obtain the subject thermo-stable enzyme, which allows the enzymatic reaction at high temperature, and is stable at high temperature and useful for resolution of optically active isomers of synthetic amino acid and analysis of terminal carboxyl group of peptide. SOLUTION: This enzyme is obtained from strains, e.g., Pyrococcus Horikosi (JCM9974), having both aminoacylase and carboxypeptidase activities, and showing the following properties: (1) optimum temperature of 90 to 95°C and optimum pH of 6.5 to 8.0; (2) causing no deactivation even under heating at 95°C for 3h (pH: 7.5); (3) molecular weight of 40,000 (SDS-PAGE); and (4) having an amino acid sequence shown by the formula. Escherichia coli BL21 (DE3) containing a gene which codes protein having the above amino acid sequence is deposited as FERM P-16051. The enzyme of the present invention can be massively produced by the DNA recombination technique. COPYRIGHT: (C)1998,JPO

Thermo-stable enzyme having aminoacylase and carboxypeptidase activity, and gene coding the same

PROBLEM TO BE SOLVED: To provide such a process in the recovery of oil and fats from rice bran as is made both to elevate a yield of rice bran oil and to increase a content of physiologically active sterol and orizanol. SOLUTION: This yield-increasing process for physiologically active rice bran oil comprises pretreating rice bran with an enzyme in the extraction of oil from the rice bran, which results in a yield of oil and fats of 20 wt.% or more on the basis of the rice bran and also a content of sterol and orizanol of 1.8 wt.% or more. The enzyme is desirably an enzyme mixture contg. amylase, cellulase, and protease.

佳次小杉

Papers

Degumming method with phospholipase

High-purity arachidonic acid triglyceride and its production

Production of oil and fat containing concentrated highly unsaturated fatty acid

Thermo-stable enzyme having aminoacylase and carboxypeptidase activity, and gene coding the same

Yield-increasing process for physiologically active rice bran oil

佳次 小杉

Papers

Degumming method with phospholipase

High-purity arachidonic acid triglyceride and its production

Production of oil and fat containing concentrated highly unsaturated fatty acid

Thermo-stable enzyme having aminoacylase and carboxypeptidase activity, and gene coding the same

Yield-increasing process for physiologically active rice bran oil

佳次小杉