WO2005097985A1 - 1,3-特異性リパーゼ粉末、その製造方法及びその使用 - Google Patents
1,3-特異性リパーゼ粉末、その製造方法及びその使用 Download PDFInfo
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- WO2005097985A1 WO2005097985A1 PCT/JP2005/006909 JP2005006909W WO2005097985A1 WO 2005097985 A1 WO2005097985 A1 WO 2005097985A1 JP 2005006909 W JP2005006909 W JP 2005006909W WO 2005097985 A1 WO2005097985 A1 WO 2005097985A1
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- Prior art keywords
- lipase
- powder
- aqueous solution
- lipase powder
- mass
- Prior art date
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- 108090001060 Lipase Proteins 0.000 title claims abstract description 121
- 239000004367 Lipase Substances 0.000 title claims abstract description 121
- 102000004882 Lipase Human genes 0.000 title claims abstract description 121
- 235000019421 lipase Nutrition 0.000 title claims abstract description 121
- 239000000843 powder Substances 0.000 title claims abstract description 61
- 238000000034 method Methods 0.000 title claims abstract description 17
- 239000007864 aqueous solution Substances 0.000 claims abstract description 24
- 238000001694 spray drying Methods 0.000 claims abstract description 19
- 239000000203 mixture Substances 0.000 claims abstract description 13
- 241000235403 Rhizomucor miehei Species 0.000 claims abstract description 7
- 239000003921 oil Substances 0.000 claims description 20
- 239000003925 fat Substances 0.000 claims description 17
- 238000005809 transesterification reaction Methods 0.000 claims description 17
- 239000002245 particle Substances 0.000 claims description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- 239000000243 solution Substances 0.000 claims description 12
- 239000012528 membrane Substances 0.000 claims description 11
- 238000004519 manufacturing process Methods 0.000 claims description 7
- 235000019198 oils Nutrition 0.000 description 16
- 235000019197 fats Nutrition 0.000 description 14
- 238000006243 chemical reaction Methods 0.000 description 9
- 230000000052 comparative effect Effects 0.000 description 8
- 238000001035 drying Methods 0.000 description 7
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 150000002148 esters Chemical class 0.000 description 5
- 238000003756 stirring Methods 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 235000019484 Rapeseed oil Nutrition 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- 239000003957 anion exchange resin Substances 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 235000014113 dietary fatty acids Nutrition 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- -1 etc. Substances 0.000 description 3
- 239000000194 fatty acid Substances 0.000 description 3
- 229930195729 fatty acid Natural products 0.000 description 3
- 150000004665 fatty acids Chemical class 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 238000005374 membrane filtration Methods 0.000 description 3
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 3
- 235000014593 oils and fats Nutrition 0.000 description 3
- 239000008363 phosphate buffer Substances 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- 239000007921 spray Substances 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- 235000000346 sugar Nutrition 0.000 description 3
- URAYPUMNDPQOKB-UHFFFAOYSA-N triacetin Chemical compound CC(=O)OCC(OC(C)=O)COC(C)=O URAYPUMNDPQOKB-UHFFFAOYSA-N 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 235000010469 Glycine max Nutrition 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 240000008042 Zea mays Species 0.000 description 2
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 2
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 2
- 239000003054 catalyst Substances 0.000 description 2
- 235000005822 corn Nutrition 0.000 description 2
- 238000005886 esterification reaction Methods 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 235000013312 flour Nutrition 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 235000013773 glyceryl triacetate Nutrition 0.000 description 2
- 235000019626 lipase activity Nutrition 0.000 description 2
- 239000004006 olive oil Substances 0.000 description 2
- 235000008390 olive oil Nutrition 0.000 description 2
- 238000010979 pH adjustment Methods 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 230000009257 reactivity Effects 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 230000000717 retained effect Effects 0.000 description 2
- 229960002622 triacetin Drugs 0.000 description 2
- VLPFTAMPNXLGLX-UHFFFAOYSA-N trioctanoin Chemical compound CCCCCCCC(=O)OCC(OC(=O)CCCCCCC)COC(=O)CCCCCCC VLPFTAMPNXLGLX-UHFFFAOYSA-N 0.000 description 2
- PHYFQTYBJUILEZ-IUPFWZBJSA-N triolein Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC(OC(=O)CCCCCCC\C=C/CCCCCCCC)COC(=O)CCCCCCC\C=C/CCCCCCCC PHYFQTYBJUILEZ-IUPFWZBJSA-N 0.000 description 2
- 235000015112 vegetable and seed oil Nutrition 0.000 description 2
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 1
- LDVVTQMJQSCDMK-UHFFFAOYSA-N 1,3-dihydroxypropan-2-yl formate Chemical compound OCC(CO)OC=O LDVVTQMJQSCDMK-UHFFFAOYSA-N 0.000 description 1
- 108090000604 Hydrolases Proteins 0.000 description 1
- 102000004157 Hydrolases Human genes 0.000 description 1
- 108010048733 Lipozyme Proteins 0.000 description 1
- 241001558145 Mucor sp. Species 0.000 description 1
- 241000235402 Rhizomucor Species 0.000 description 1
- 241000135252 Rhizomucor sp. Species 0.000 description 1
- 235000019485 Safflower oil Nutrition 0.000 description 1
- 229930182558 Sterol Natural products 0.000 description 1
- BAECOWNUKCLBPZ-HIUWNOOHSA-N Triolein Natural products O([C@H](OCC(=O)CCCCCCC/C=C\CCCCCCCC)COC(=O)CCCCCCC/C=C\CCCCCCCC)C(=O)CCCCCCC/C=C\CCCCCCCC BAECOWNUKCLBPZ-HIUWNOOHSA-N 0.000 description 1
- PHYFQTYBJUILEZ-UHFFFAOYSA-N Trioleoylglycerol Natural products CCCCCCCCC=CCCCCCCCC(=O)OCC(OC(=O)CCCCCCCC=CCCCCCCCC)COC(=O)CCCCCCCC=CCCCCCCCC PHYFQTYBJUILEZ-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 150000008044 alkali metal hydroxides Chemical class 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- OEERIBPGRSLGEK-UHFFFAOYSA-N carbon dioxide;methanol Chemical compound OC.O=C=O OEERIBPGRSLGEK-UHFFFAOYSA-N 0.000 description 1
- 150000001733 carboxylic acid esters Chemical class 0.000 description 1
- 150000001735 carboxylic acids Chemical class 0.000 description 1
- 239000012159 carrier gas Substances 0.000 description 1
- 239000003729 cation exchange resin Substances 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 229920001429 chelating resin Polymers 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000003240 coconut oil Substances 0.000 description 1
- 235000019864 coconut oil Nutrition 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 235000005687 corn oil Nutrition 0.000 description 1
- 239000002285 corn oil Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000032050 esterification Effects 0.000 description 1
- DJLUGWFUVRDHLO-UHFFFAOYSA-N ethyl 4,5-dimethyl-6-oxo-7-propyl-7,8-dihydrocyclopenta[e][1]benzofuran-2-carboxylate Chemical class O=C1C(CCC)CC2=C1C(C)=C(C)C1=C2C=C(C(=O)OCC)O1 DJLUGWFUVRDHLO-UHFFFAOYSA-N 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 239000001087 glyceryl triacetate Substances 0.000 description 1
- 239000010651 grapefruit oil Substances 0.000 description 1
- 239000001307 helium Substances 0.000 description 1
- 229910052734 helium Inorganic materials 0.000 description 1
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 description 1
- 239000012510 hollow fiber Substances 0.000 description 1
- 239000012051 hydrophobic carrier Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- FCCDDURTIIUXBY-UHFFFAOYSA-N lipoamide Chemical compound NC(=O)CCCCC1CCSS1 FCCDDURTIIUXBY-UHFFFAOYSA-N 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000004570 mortar (masonry) Substances 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 229940049964 oleate Drugs 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 229920002492 poly(sulfone) Polymers 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 235000005713 safflower oil Nutrition 0.000 description 1
- 239000003813 safflower oil Substances 0.000 description 1
- 238000005185 salting out Methods 0.000 description 1
- 239000008159 sesame oil Substances 0.000 description 1
- 235000011803 sesame oil Nutrition 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 239000003549 soybean oil Substances 0.000 description 1
- 235000012424 soybean oil Nutrition 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 235000003702 sterols Nutrition 0.000 description 1
- 150000005846 sugar alcohols Polymers 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 150000003626 triacylglycerols Chemical class 0.000 description 1
- 229940093609 tricaprylin Drugs 0.000 description 1
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 1
- 229940117972 triolein Drugs 0.000 description 1
- 235000019871 vegetable fat Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
Classifications
-
- 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
- C12P7/6445—Glycerides
- C12P7/6458—Glycerides by transesterification, e.g. interesterification, ester interchange, alcoholysis or acidolysis
-
- 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
-
- 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
Definitions
- the present invention relates to a 1,3-specific lipase powder which can be suitably used for transesterification of various fats and oils, a method for producing the same, and a method for immersing or infiltrating a 1,3-specific lipase powder in fat or oil.
- the present invention relates to a lipase composition, a method for transesterifying fats and oils using the lipase powder, and the like.
- Lipases are widely used in esterification reactions between various carboxylic acids such as fatty acids and alcohols such as monoalcohols and polyhydric alcohols, and transesterification reactions between a plurality of carboxylic acid esters. Of these, transesterification is an important technology for the production of esters, sugar esters and steroids of various fatty acids, including the modification of animal and vegetable fats and oils. If lipase, which is a fat and oil hydrolase, is used as a catalyst for these reactions, transesterification can be carried out under mild conditions at room temperature to about 70 ° C. The lipase as a catalyst is a natural product, and its safety is also high. In addition, the target product can be efficiently produced due to its substrate specificity and positional specificity.
- lipase has conventionally been used as a carrier such as an anion exchange resin (Patent Document 1), a phenol-adsorbed resin (Patent Document 2), a hydrophobic carrier (Patent Document 3), and a cation exchange resin (Patent Document 1). 4), immobilized on chelating resins (Patent Document 5) and the like, and used for esterification and ester exchange reactions.
- a carrier such as an anion exchange resin (Patent Document 1), a phenol-adsorbed resin (Patent Document 2), a hydrophobic carrier (Patent Document 3), and a cation exchange resin (Patent Document 1). 4), immobilized on chelating resins (Patent Document 5) and the like, and used for esterification and ester exchange reactions.
- lipase is immobilized and used for the transesterification reaction.
- a powerful immobilized lipase is not only accompanied by loss of the original lipase activity due to the immobilization process, but also has a porous structure.
- an acidic carrier was used, the pores were clogged with raw materials and products, resulting in a decrease in the transesterification rate.
- water retained by the carrier is brought into the reaction system, so that a side reaction, for example, It was difficult to avoid the production of diglyceride / monoglyceride in the transesterification of oils and fats.
- a lipase is added to the ester-containing raw material so that 90% or more of the dispersed lipase powder particles are maintained at a particle size in the range of 1 to 100 ⁇ m during the transesterification reaction.
- a method of performing a transesterification reaction by dispersing a powder has been proposed (Patent Document 6). It has also been proposed to use an enzyme powder obtained by drying an enzyme solution containing a phospholipid and a fat-soluble vitamin (Patent Document 7).
- Patent Document 1 Japanese Patent Application Laid-Open No. Sho 60-98984
- Patent Document 2 JP-A-61-202688
- Patent document 3 JP-A-2-138986
- Patent Document 4 JP-A-3-61485
- Patent Document 5 JP-A-1-2622795
- Patent Document 6 Patent No. 2668187
- Patent Document 7 JP-A-2000-106873
- An object of the present invention is to provide a lipase powder in which 1,3-selectivity of a 1,3-specific lipase is improved.
- Another object of the present invention is to provide a lipase composition obtained by immersing or infiltrating the lipase powder in fat or oil.
- the 1,3-specific lipase derived from Rhizomucor miehei is a lipase with extremely high 1,3 selectivity and is used by being immobilized on an anion exchange resin.
- the present invention has been made based on the finding that a spherical powder having a water content of 10% by mass or less, without using it, further improves the 1,3 selectivity!
- the present invention provides a 1,3-specific lipase derived from Rhizomucor miehe oil, which is spherical and has a water content of 10% by mass or less.
- the present invention also provides a lipase composition wherein the lipase powder is immersed or infiltrated in fat or oil.
- the present invention also provides a method for producing the lipase powder, which comprises spray-drying a lipase-containing aqueous solution whose pH has been adjusted to 6 to 7.5.
- the present invention also provides a transesterification lipase containing the lipase powder.
- the present invention also provides a method for ester exchange of fats and oils, which comprises using the above-mentioned lipase for transesterification.
- the 1,3-specific lipase targeted in the present invention is a 1,3-specific lipase derived from Rhizomucor miehei belonging to the genus Rhizomucor sp. Conventionally, Rhizomucor miehei; is sometimes treated as a genus Mucor sp.
- this lipase is used as a powder having a spherical shape and a water content of 10% by mass or less.
- the water content is preferably from 6.5 to 8.5% by weight to a force of not more than 10% by weight.
- the particle size of the lipase powder of the present invention can be arbitrarily set, it is preferable that 90% by mass or more of the lipase powder has a particle size of 1 to 100 ⁇ m.
- the average particle size is preferably from 20 to 80 ⁇ m, more preferably from 20 to 50 ⁇ m.
- the particle size of the lipase powder can be measured, for example, using a particle size distribution analyzer (LA-500) manufactured by HORIBA.
- LA-500 particle size distribution analyzer manufactured by HORIBA.
- Such a lipase powder can be easily prepared by, for example, spray-drying (spray drying) an aqueous solution containing a lipase.
- the lipase-containing aqueous solution includes a lipase culture solution from which cells have been removed, a purified culture solution, a solution obtained by dissolving and dispersing a lipase powder obtained from these in water, and a commercially available lipase solution.
- examples thereof include those in which the powder is dissolved and dispersed again in water, commercially available liquid lipase, and the like.
- those from which low-molecular components such as salts are removed to further enhance lipase activity are preferable, and those from which low-molecular components such as sugar are removed to further improve powder properties are more preferable.
- Lipase cultures include, for example, soy flour, peptone, corn 'Stap' liquor, KH
- Aqueous solution containing PO, (NH) SO, MgSO containing PO, (NH) SO, MgSO.
- soy flour 0.1 to 20 wt% preferably from 1.0 to 10 wt%, peptone 0.1 to 30 mass 0/0, preferably from 0.5 to 10 weight 0/0, corn ' Suthep 'liquor from 0.1 to 30 weight 0/0, preferably rather is from 0.5 to 10 weight 0/0, ⁇ ⁇ 0. 01 ⁇ 20 mass 0/0, preferably from 0.1 to 5 mass 0/0
- soy flour 0.1 to 20 wt% preferably from 1.0 to 10 wt%, peptone 0.1 to 30 mass 0/0, preferably from 0.5 to 10 weight 0/0, corn ' Suthep 'liquor from 0.1 to 30 weight 0/0, preferably rather is from 0.5 to 10 weight 0/0, ⁇ ⁇ 0. 01 ⁇ 20 mass 0/0, preferably from 0.1 to 5 mass 0/0
- peptone 0.1 to 30 mass 0/0 preferably from 0.5 to 10 weight 0/0
- ⁇ is 0.01 to 20% by mass, preferably 0.05 to 5% by mass.
- Culture conditions are culture temperature
- Isolation of the cells is preferably performed by centrifugation, membrane filtration, or the like. Further, removal of low molecular components such as salts and sugars can be performed by UF membrane treatment. Specifically, after performing UF membrane treatment, concentrating the aqueous solution containing lipase to a volume of 1Z2, and then adding the same amount of phosphate buffer as the concentrated solution, it is repeated 1 to 5 times. A lipase-containing aqueous solution from which low molecular components have been removed can be obtained.
- the centrifugation is preferably controlled at 200 to 20,000 X g, and the membrane filtration is preferably controlled at a pressure of 3.0 kg / m2 or less using an MF membrane, a filter press or the like.
- the stirring speed of the homogenizer is 500 to 30,000 rpm, preferably ⁇ 1,000 to 15,000 rpm, and the rotating speed of the worst blender is 500 to 10,000, rpm, preferably ⁇ 1,000 to 5, OOOrpm. is there.
- the stirring time is 0.5 to 10 minutes, preferably 1 to 5 minutes.
- the ultrasonic crushing is preferably performed under the conditions of 1 to 50 KHz, preferably 10 to 20 KHz. It is better to use glass balls having a diameter of about 0.1 to 0.5 mm for the ball mill. In the present invention, it is preferable to use an aqueous solution containing 5 to 30% by mass as a solid content as the lipase-containing aqueous solution.
- the solid content concentration in the lipase-containing aqueous solution can be determined as Brix.
- the pH of the lipase-containing aqueous solution is preferably adjusted to 6 to 7.5.
- pH adjustment may be performed in any step before the drying step such as spray drying (spray drying) so that the pH of the lipase-containing aqueous solution is adjusted in advance so that the pH immediately before the drying step is within the above range. It may be adjusted.
- an alkali metal hydroxide such as sodium hydroxide which can use various alkali agents and acids.
- the lipase-containing aqueous solution may be concentrated in a middle step before the drying step.
- the method of concentration is not particularly limited, but evaporator, flash evaporator, UF membrane concentration, MF membrane concentration, salting out with inorganic salts, precipitation method with a solvent, adsorption method with ion exchange cellulose, etc., water absorbing gel Water absorption method.
- UF membrane concentration and evaporator are used.
- Spray drying is preferably performed using a spray dryer such as a nozzle countercurrent type, a disk countercurrent type, a nozzle cocurrent type, and a disk cocurrent type.
- a spray dryer such as a nozzle countercurrent type, a disk countercurrent type, a nozzle cocurrent type, and a disk cocurrent type.
- Preferably may Disk parallel flow force s, atomizer revolution number 4, 000 ⁇ 20, OOOrpm, Caro heat population temperature 100 to 20 0 ° C, outlet temperature 40 - control to spray drying in L00 ° C ( Spray drying).
- the lipase powder thus prepared can be used as it is. It is handled as a lipase composition immersed or infiltrated in fats and oils.
- the mass of the fat or oil in the lipase composition is preferably 0.1 to 20 times, more preferably 1 to 20 times, the lipase powder.
- the lipase composition is prepared by adding oil to a lipase powder produced by spray drying (spray drying) or the like, and uniformly stirring the mixture with a stirrer, a three-one motor or the like.
- the fats and oils can be easily obtained by adding fats and oils to the powder recovery section of the laser dryer in advance, stirring uniformly after recovery, and removing excess fats and oils by filtration.
- oils and fats to be impregnated or infiltrated into the lipase powder are not particularly limited, and vegetable oils such as rapeseed oil, soybean oil, grapefruit oil, olive oil, olive oil, safflower oil, corn oil, coconut oil, sesame oil, etc.
- vegetable oils such as rapeseed oil, soybean oil, grapefruit oil, olive oil, olive oil, safflower oil, corn oil, coconut oil, sesame oil, etc.
- triacylglycerols such as triolein (glycerin trioleate), tricaprylin (glycerin trioctanoate), triacetin (glycerin triacetate), triptyline (glycerin triptylate), fatty acid esters, and sterol esters.
- Triacylglycerols such as triolein (glycerin trioleate), tricaprylin (glycerin trioctanoate), triacetin (glycerin triacetate), tripty
- the 1,3 selectivity of 1,3 specific lipase is improved in each step, and the fatty acid residue at position 2 of the starting triglyceride can be retained in the transesterified product at a very high ratio. it can.
- a lipase (product name: Palatase 20000L) derived from Rhizomucor miehei manufactured by Novozyms Japan Ltd. in a form in which lipase is dissolved and dispersed in an aqueous solution is used by using a UF module (Asahi Kasei Kogyo Co., Ltd. SIP-0013). A low-molecular component was removed to obtain a lipase-containing aqueous solution (solid content: 10.6% by mass).
- liquid lipase (paratase 20000 L) was treated with a UF membrane under ice-cooling, concentrated to a volume of 1Z2, and then 0.01 M phosphate buffer of pH 7 was added in the same amount as the concentrated solution. The same operation of adding the phosphate buffer to the UF membrane after treatment of the obtained solution was repeated three times to obtain a lipase-containing aqueous solution (lipase concentrated solution: buffer at a volume ratio of 1: 1).
- the pH of the lipase-containing aqueous solution was adjusted to pH 6.8 to 6.9 using an aqueous sodium hydroxide solution.
- This liquid is sprayed using a spray dryer (SD-1000: manufactured by Tokyo Rika Kikai Co., Ltd.) under the conditions of an inlet temperature of 130 ° C, a dry air volume of 0.7 to 1. lmVmin, and a spray pressure of ll to 12 kpa. Then Pase powder was obtained.
- the shape of the lipase powder particles was spherical, and 90% by mass or more of the lipase powder was in the range of particle size 1 to LOO ⁇ m, and the average particle size was 8.2 ⁇ m.
- the particle size was measured using a particle size distribution analyzer (LA-500) manufactured by HORIBA. The water content measured by the method of drying at 105 ° C for 1 hour and heat was 7.9% by mass.
- the solid content concentration in the lipase-containing aqueous solution was determined as Brix.% Using a saccharimeter (BRX-242, manufactured by CIS Co., Ltd.).
- Lipozyme RMIM manufactured by Novozims Japan KK in which lipase derived from Rhizomucor miehei was immobilized on an anion exchange resin was used as Comparative Example 1.
- a lipase powder was obtained in the same manner as in Example 1, except that freeze drying was performed instead of spray drying (spray drying).
- freeze-drying a lipase-containing aqueous solution whose pH has been adjusted to 6.8 to 6.9 is placed in an eggplant-shaped flask, frozen with dry ice methanol, and then freeze-dried by Tokyo Rika Kikai Co., Ltd. (FDU-830). And freeze-dried at 0.15 Torr for 1 day. After drying, the mixture was lightly ground in a mortar to obtain a lipase powder.
- the shape of the obtained lipase powder was amorphous, and the water content was 11.3% by mass.
- Example 1,3-selectivity of the lipase powders of Comparative Example 1 and Comparative Example 2 was measured by the following method.
- Table 1 shows the measurement results of 1,3-selectivity of Example 1 and Comparative Example 1.
- the lipase powder obtained in Comparative Example 2 had a strong transesterification activity.
- reaction substrates Using lmol of GRYCERYL-1,3-PALMITATE-2-OLEATE (POP) and 3mol of OCTANOIC ETHYL (C8Et) as reaction substrates, and adding lipase powder to be 0.5 to 5% by weight of the substrate, depending on the enzyme activity, The reaction was performed at 60 ° C, sampled over time, and diluted with hexane. This sample was analyzed by GC, and the reaction rates at the 1st and 3rd positions (C16: 0Et) and 2nd position (C18: 1Et) were determined by the following formula.
- the value of the final reaction rate at this time is variable.
- the reactivity of the 1,3-positions when the reactivity of the 2-position was set to 1 was determined.
- Carrier gas helium
- a 5-fold amount of rapeseed oil was added to the lipase powder obtained in Example 1, the lipase powder was impregnated with the rapeseed oil, and the oil and fat was removed by filtration.
- the weight of lipase powder / rapeseed oil was 55/45. Was prepared.
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Priority Applications (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CA002530200A CA2530200A1 (en) | 2004-04-08 | 2005-04-08 | 1,3-specific lipase powder, methods for producing the same and use thereof |
EP05728498A EP1734115A4 (en) | 2004-04-08 | 2005-04-08 | POWDER OF A 1,3-SPECIFIC LIPASE, METHOD FOR THE PRODUCTION THEREOF AND USE THEREOF |
CN2005800005485A CN1806044B (zh) | 2004-04-08 | 2005-04-08 | 1,3-特异性脂肪酶粉末、其制备方法及其应用 |
JP2006519483A JPWO2005097985A1 (ja) | 2004-04-08 | 2005-04-08 | 1,3−特異性リパーゼ粉末、その製造方法及びその使用 |
US11/320,760 US20060105438A1 (en) | 2004-04-08 | 2005-12-30 | 1,3-Specific lipase powder, methods for producing the same and use thereof |
Applications Claiming Priority (2)
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JP2004-114444 | 2004-04-08 | ||
JP2004114444 | 2004-04-08 |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
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US11/320,760 Continuation US20060105438A1 (en) | 2004-04-08 | 2005-12-30 | 1,3-Specific lipase powder, methods for producing the same and use thereof |
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WO2005097985A1 true WO2005097985A1 (ja) | 2005-10-20 |
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PCT/JP2005/006909 WO2005097985A1 (ja) | 2004-04-08 | 2005-04-08 | 1,3-特異性リパーゼ粉末、その製造方法及びその使用 |
Country Status (9)
Country | Link |
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US (1) | US20060105438A1 (ja) |
EP (1) | EP1734115A4 (ja) |
JP (1) | JPWO2005097985A1 (ja) |
KR (1) | KR20070006657A (ja) |
CN (1) | CN1806044B (ja) |
CA (1) | CA2530200A1 (ja) |
MY (1) | MY155007A (ja) |
TW (1) | TW200538464A (ja) |
WO (1) | WO2005097985A1 (ja) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2011068076A1 (ja) * | 2009-12-01 | 2011-06-09 | 日清オイリオグループ株式会社 | リパーゼ粉末製剤及びその使用 |
JP5145609B2 (ja) * | 2007-03-16 | 2013-02-20 | 日清オイリオグループ株式会社 | リパーゼ粉末製剤、その製造方法及び使用 |
CN108165540A (zh) * | 2018-02-11 | 2018-06-15 | 中国农业大学 | 一种米黑根毛霉α-淀粉酶及其编码基因与应用 |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE102004019472A1 (de) * | 2004-04-22 | 2005-11-17 | Bayer Healthcare Ag | Phenylacetamide |
UA97127C2 (uk) * | 2006-12-06 | 2012-01-10 | Бандж Ойлз, Инк. | Спосіб безперервної ферментативної обробки композиції, що містить ліпід, та система для його здійснення |
Citations (2)
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JPS5417179A (en) * | 1977-07-08 | 1979-02-08 | Sankyo Co Ltd | Preparation of powdered enzyme |
JPH0779789A (ja) * | 1993-09-17 | 1995-03-28 | Nisshin Oil Mills Ltd:The | リパーゼ粉末を用いたエステル交換法 |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
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US4233405A (en) * | 1979-10-10 | 1980-11-11 | Rohm And Haas Company | Process for spray drying enzymes |
DK402583D0 (da) * | 1983-09-05 | 1983-09-05 | Novo Industri As | Fremgangsmade til fremstilling af et immobiliseret lipasepraeparat og anvendelse deraf |
US5288619A (en) * | 1989-12-18 | 1994-02-22 | Kraft General Foods, Inc. | Enzymatic method for preparing transesterified oils |
CN1181187C (zh) * | 2003-04-22 | 2004-12-22 | 江南大学 | 一种脂肪酶产生菌及其筛选方法和产业化应用 |
-
2005
- 2005-04-07 TW TW094111045A patent/TW200538464A/zh unknown
- 2005-04-07 MY MYPI20051553A patent/MY155007A/en unknown
- 2005-04-08 WO PCT/JP2005/006909 patent/WO2005097985A1/ja not_active Application Discontinuation
- 2005-04-08 CN CN2005800005485A patent/CN1806044B/zh active Active
- 2005-04-08 JP JP2006519483A patent/JPWO2005097985A1/ja active Pending
- 2005-04-08 EP EP05728498A patent/EP1734115A4/en not_active Withdrawn
- 2005-04-08 KR KR1020067000996A patent/KR20070006657A/ko not_active Application Discontinuation
- 2005-04-08 CA CA002530200A patent/CA2530200A1/en not_active Abandoned
- 2005-12-30 US US11/320,760 patent/US20060105438A1/en not_active Abandoned
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS5417179A (en) * | 1977-07-08 | 1979-02-08 | Sankyo Co Ltd | Preparation of powdered enzyme |
JPH0779789A (ja) * | 1993-09-17 | 1995-03-28 | Nisshin Oil Mills Ltd:The | リパーゼ粉末を用いたエステル交換法 |
Non-Patent Citations (3)
Title |
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DECAGNY B. ET AL.: "1H-NMR on line monitoring of water activity during lipase catalysed esterification.", BIOCHIM.BIOPHYS.ACTA., vol. 1387, no. 1-2, 1998, pages 129 - 135, XP004278467 * |
See also references of EP1734115A4 * |
XIAO Y. ET AL.: "Purification and partial characterization of Rhizomucor meihei lipase for ester synthesis.", APPL.BIOCHEM.BIOTECH., vol. 59, no. 2, 1996, pages 145 - 158, XP008052456 * |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP5145609B2 (ja) * | 2007-03-16 | 2013-02-20 | 日清オイリオグループ株式会社 | リパーゼ粉末製剤、その製造方法及び使用 |
WO2011068076A1 (ja) * | 2009-12-01 | 2011-06-09 | 日清オイリオグループ株式会社 | リパーゼ粉末製剤及びその使用 |
EP2508598A1 (en) * | 2009-12-01 | 2012-10-10 | The Nisshin OilliO Group, Ltd. | Lipase powder preparation and use thereof |
EP2508598A4 (en) * | 2009-12-01 | 2013-05-29 | Nisshin Oillio Group Ltd | PREPARATION OF LIPASE POWDER AND USE THEREOF |
CN108165540A (zh) * | 2018-02-11 | 2018-06-15 | 中国农业大学 | 一种米黑根毛霉α-淀粉酶及其编码基因与应用 |
CN108165540B (zh) * | 2018-02-11 | 2020-09-01 | 中国农业大学 | 一种米黑根毛霉α-淀粉酶及其编码基因与应用 |
Also Published As
Publication number | Publication date |
---|---|
JPWO2005097985A1 (ja) | 2008-02-28 |
EP1734115A1 (en) | 2006-12-20 |
CN1806044A (zh) | 2006-07-19 |
EP1734115A4 (en) | 2007-10-24 |
TW200538464A (en) | 2005-12-01 |
US20060105438A1 (en) | 2006-05-18 |
CN1806044B (zh) | 2012-01-11 |
KR20070006657A (ko) | 2007-01-11 |
CA2530200A1 (en) | 2005-10-20 |
MY155007A (en) | 2015-08-28 |
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