CN105177073A - Method of preparing phosphatidylserine - Google Patents
Method of preparing phosphatidylserine Download PDFInfo
- Publication number
- CN105177073A CN105177073A CN201510566411.1A CN201510566411A CN105177073A CN 105177073 A CN105177073 A CN 105177073A CN 201510566411 A CN201510566411 A CN 201510566411A CN 105177073 A CN105177073 A CN 105177073A
- Authority
- CN
- China
- Prior art keywords
- solution
- phosphatidylserine
- lecithin materials
- chain fat
- acetoacetic ester
- Prior art date
- Legal status (The legal status 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 status listed.)
- Granted
Links
- TZCPCKNHXULUIY-RGULYWFUSA-N 1,2-distearoyl-sn-glycero-3-phosphoserine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(=O)OC[C@H](N)C(O)=O)OC(=O)CCCCCCCCCCCCCCCCC TZCPCKNHXULUIY-RGULYWFUSA-N 0.000 title claims abstract description 69
- ZWZWYGMENQVNFU-UHFFFAOYSA-N Glycerophosphorylserin Natural products OC(=O)C(N)COP(O)(=O)OCC(O)CO ZWZWYGMENQVNFU-UHFFFAOYSA-N 0.000 title claims abstract description 69
- 238000000034 method Methods 0.000 title claims abstract description 55
- 238000006243 chemical reaction Methods 0.000 claims abstract description 45
- 102000011420 Phospholipase D Human genes 0.000 claims abstract description 29
- 108090000553 Phospholipase D Proteins 0.000 claims abstract description 29
- 238000000199 molecular distillation Methods 0.000 claims abstract description 29
- 150000003904 phospholipids Chemical class 0.000 claims abstract description 22
- 241001465754 Metazoa Species 0.000 claims abstract description 20
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 claims abstract description 4
- 239000000243 solution Substances 0.000 claims description 79
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 claims description 56
- 239000000787 lecithin Substances 0.000 claims description 56
- 229940067606 lecithin Drugs 0.000 claims description 56
- 235000010445 lecithin Nutrition 0.000 claims description 56
- 239000000463 material Substances 0.000 claims description 51
- XYIBRDXRRQCHLP-UHFFFAOYSA-N ethyl acetoacetate Chemical compound CCOC(=O)CC(C)=O XYIBRDXRRQCHLP-UHFFFAOYSA-N 0.000 claims description 37
- 239000012071 phase Substances 0.000 claims description 34
- 238000004821 distillation Methods 0.000 claims description 26
- 239000011259 mixed solution Substances 0.000 claims description 26
- 235000019198 oils Nutrition 0.000 claims description 20
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 claims description 16
- 239000000203 mixture Substances 0.000 claims description 16
- 238000003756 stirring Methods 0.000 claims description 16
- 239000008351 acetate buffer Substances 0.000 claims description 14
- 238000010438 heat treatment Methods 0.000 claims description 12
- 125000004494 ethyl ester group Chemical group 0.000 claims description 11
- 239000008346 aqueous phase Substances 0.000 claims description 10
- 238000013016 damping Methods 0.000 claims description 10
- YYZUSRORWSJGET-UHFFFAOYSA-N ethyl octanoate Chemical compound CCCCCCCC(=O)OCC YYZUSRORWSJGET-UHFFFAOYSA-N 0.000 claims description 10
- 239000012530 fluid Substances 0.000 claims description 10
- 235000021323 fish oil Nutrition 0.000 claims description 9
- LVGKNOAMLMIIKO-UHFFFAOYSA-N Elaidinsaeure-aethylester Natural products CCCCCCCCC=CCCCCCCCC(=O)OCC LVGKNOAMLMIIKO-UHFFFAOYSA-N 0.000 claims description 7
- LVGKNOAMLMIIKO-QXMHVHEDSA-N ethyl oleate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC LVGKNOAMLMIIKO-QXMHVHEDSA-N 0.000 claims description 7
- 229940093471 ethyl oleate Drugs 0.000 claims description 7
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 6
- 239000001110 calcium chloride Substances 0.000 claims description 6
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 6
- 230000035484 reaction time Effects 0.000 claims description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 abstract description 27
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 abstract description 19
- 239000007788 liquid Substances 0.000 abstract description 14
- 238000009835 boiling Methods 0.000 abstract description 12
- 238000004519 manufacturing process Methods 0.000 abstract description 10
- 239000003960 organic solvent Substances 0.000 abstract description 6
- 238000003912 environmental pollution Methods 0.000 abstract description 5
- 230000003139 buffering effect Effects 0.000 abstract 1
- 238000009776 industrial production Methods 0.000 abstract 1
- 231100000331 toxic Toxicity 0.000 abstract 1
- 230000002588 toxic effect Effects 0.000 abstract 1
- 102000004190 Enzymes Human genes 0.000 description 11
- 108090000790 Enzymes Proteins 0.000 description 11
- 102000002322 Egg Proteins Human genes 0.000 description 8
- 108010000912 Egg Proteins Proteins 0.000 description 8
- 239000000284 extract Substances 0.000 description 8
- 210000004681 ovum Anatomy 0.000 description 8
- 239000002904 solvent Substances 0.000 description 7
- 230000002051 biphasic effect Effects 0.000 description 6
- DVSZKTAMJJTWFG-UHFFFAOYSA-N docosa-2,4,6,8,10,12-hexaenoic acid Chemical compound CCCCCCCCCC=CC=CC=CC=CC=CC=CC(O)=O DVSZKTAMJJTWFG-UHFFFAOYSA-N 0.000 description 6
- MBMBGCFOFBJSGT-KUBAVDMBSA-N docosahexaenoic acid Natural products CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCC(O)=O MBMBGCFOFBJSGT-KUBAVDMBSA-N 0.000 description 6
- 239000000843 powder Substances 0.000 description 6
- 239000002360 explosive Substances 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 238000006555 catalytic reaction Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 230000001131 transforming effect Effects 0.000 description 4
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- 210000004556 brain Anatomy 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 239000007789 gas Substances 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 208000024827 Alzheimer disease Diseases 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000012159 carrier gas Substances 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 239000007791 liquid phase Substances 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 125000001820 oxy group Chemical group [*:1]O[*:2] 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 2
- HNAGHMKIPMKKBB-UHFFFAOYSA-N 1-benzylpyrrolidine-3-carboxamide Chemical compound C1C(C(=O)N)CCN1CC1=CC=CC=C1 HNAGHMKIPMKKBB-UHFFFAOYSA-N 0.000 description 1
- HIXDQWDOVZUNNA-UHFFFAOYSA-N 2-(3,4-dimethoxyphenyl)-5-hydroxy-7-methoxychromen-4-one Chemical compound C=1C(OC)=CC(O)=C(C(C=2)=O)C=1OC=2C1=CC=C(OC)C(OC)=C1 HIXDQWDOVZUNNA-UHFFFAOYSA-N 0.000 description 1
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 208000031295 Animal disease Diseases 0.000 description 1
- 208000006096 Attention Deficit Disorder with Hyperactivity Diseases 0.000 description 1
- 208000036864 Attention deficit/hyperactivity disease Diseases 0.000 description 1
- 241000238366 Cephalopoda Species 0.000 description 1
- 241000283074 Equus asinus Species 0.000 description 1
- 241000283073 Equus caballus Species 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 206010039966 Senile dementia Diseases 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-N acetic acid Substances CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 210000001557 animal structure Anatomy 0.000 description 1
- 238000000149 argon plasma sintering Methods 0.000 description 1
- PKSROMPNLONTJT-UHFFFAOYSA-N azanium;chloroform;methanol;hydroxide Chemical group N.O.OC.ClC(Cl)Cl PKSROMPNLONTJT-UHFFFAOYSA-N 0.000 description 1
- 210000004958 brain cell Anatomy 0.000 description 1
- 230000006931 brain damage Effects 0.000 description 1
- 231100000874 brain damage Toxicity 0.000 description 1
- 230000003925 brain function Effects 0.000 description 1
- 208000029028 brain injury Diseases 0.000 description 1
- OBNCKNCVKJNDBV-UHFFFAOYSA-N butanoic acid ethyl ester Natural products CCCC(=O)OCC OBNCKNCVKJNDBV-UHFFFAOYSA-N 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 241001233037 catfish Species 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000033077 cellular process Effects 0.000 description 1
- WORJEOGGNQDSOE-UHFFFAOYSA-N chloroform;methanol Chemical compound OC.ClC(Cl)Cl WORJEOGGNQDSOE-UHFFFAOYSA-N 0.000 description 1
- 208000012839 conversion disease Diseases 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 230000032050 esterification Effects 0.000 description 1
- 238000005886 esterification reaction Methods 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- PEDCQBHIVMGVHV-UHFFFAOYSA-N glycerol group Chemical group OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 1
- 239000004519 grease Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 235000001497 healthy food Nutrition 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000036651 mood Effects 0.000 description 1
- 210000003061 neural cell Anatomy 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 238000010606 normalization Methods 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 230000007096 poisonous effect Effects 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000004853 protein function Effects 0.000 description 1
- 239000010453 quartz Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000007634 remodeling Methods 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- 239000007901 soft capsule Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
- 238000010792 warming Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to a method of preparing phosphatidylserine. The method comprises the following steps: taking phospholipase D to catalyze a basic group exchange reaction of marine animal phospholipid and L-serine; centrifuging and carrying out molecular distillation, and separating and purifying to prepare the phosphatidylserine. By utilizing a liquid-liquid double-phase reaction system formed by middle-chain and long-chain fatty acid ethyl esters and a buffering solution, and utilizing the middle-chain and long-chain fatty acid ethyl esters which have high boiling points and are safe to eat to replace the organic solvents including diethyl ether, ethyl acetate and the like which have low boiling points, are flammable and combustible and are toxic and harmful, the production safety is improved while the reaction efficiency is guaranteed and the environmental pollution is not caused, so that the method is suitable for industrial production.
Description
Technical field:
The present invention relates to field of marine biotechnology, particularly relate to a kind of method preparing phosphatidylserine.
Background technology:
Phosphatidylserine (PS) be a kind of uniquely can the phosphatide of regulating cell film key protein function, important regulating effect is had to many cellular process, its major function to improve brain cell activity, to treatment encephalatrophy, prevent senile dementia, improve the brain function of the elderly and have good curative effect, simultaneously to the effect of repairing brain damage, treatment childhood hyperkinetic syndrome has highly significant; In addition, PS can promote recovery, balance mood, the alleviate depression of brain fag.
Phosphatidylserine is present in the microbial film of all animals, higher plant and microorganism, is one of important component of cell membrane phospholipid.Containing a large amount of phosphatidylserine in human neural cells film, containing abundant DHA (docosahexenoic acid) on its glycerol backbone, to the maintenance of neurocyte, reparation and function, there is vital role.Phosphatidylserine have passed U.S. FDA certification in 2007, one of new resource food that the Ministry of Health of Ye Shi China announces for 2010.
It has been generally acknowledged that to have with DHA the physiological function better improving alzheimer's disease with the phosphatidylserine of chemical bonds, it is normally to derive from the phosphatide of marine animal for raw material.
The preparation method of phosphatidylserine mainly contains extraction method and enzyme transforming process, wherein, extraction method mainly extracts PS from vegetable cell, animal Yelkin TTS, PS content in plant is less, so extraction method is to be extracted as master in the Yelkin TTS of animal, from the Yelkin TTS of animal, extract PS mainly extract from the brain of animal and internal organ, at present, external major part with the animal brain of the poultry such as ox, sheep, rabbit, horse, donkey for raw material extracts PS.In recent years, due to Animal diseases such as mad cow diseases, the security utilizing animal Yelkin TTS to extract the method for PS is subject to the query of people.
Enzyme transforming process mainly with natural Yelkin TTS for matrix, add Serine, under the effect of Phospholipase D, generate phosphatidylserine.In prior art, the liquid liquid biphasic reaction system of the many employings of enzyme' s catalysis " organic phase-damping fluid " of phosphatidylserine, uses the organic solvents such as chloroform, benzene, ethyl butyrate, ethyl acetate or ether can obtain higher reaction conversion ratio usually.But because above-mentioned organic solvent boiling point is very low, inflammable and explosive, poisonous and harmful, easily causes environmental pollution, is unsuitable for suitability for industrialized production.
Summary of the invention:
The present invention is in order to make up the deficiencies in the prior art, provide a kind of method preparing phosphatidylserine, first react with the Baseexchange of Phospholipase D catalysis marine animal phosphatide and Serine, then through centrifugal and molecular distillation, obtained phosphatidylserine after separation and purification; The liquid liquid biphasic reaction system that in utilization, long-chain fat acetoacetic ester and damping fluid are formed, long-chain fat acetoacetic ester substitutes low, the inflammable and explosive and virulent organic solvent of the boiling point such as ether and ethyl acetate with high boiling point and in edible safety, production security is improve while ensure that reaction efficiency, avoid causing environmental pollution, be suitable for suitability for industrialized production, efficiently solve problems of the prior art.
The present invention for solving the problems of the technologies described above adopted technical scheme is:
Prepare a method for phosphatidylserine, comprise the following steps:
(1) from marine animal, lecithin materials is extracted to obtain;
(2) lecithin materials obtained in step (1) is dissolved in middle long-chain fat acetoacetic ester, obtains solution A;
(3) Phospholipase D and Serine being dissolved in pH value is in the acetate buffer solution of 4.5 ~ 6.0, obtains solution B;
(4) solution A obtained in step (2) is added in the solution B obtained in step (3), obtain mixed solution C, the mixed solution C heating obtained, stirring reaction, obtain reacted solution D;
(5) adopt centrifugal method to remove aqueous phase the reacted solution D obtained in step (4) and obtain oil phase;
(6) adopt molecular distillation method to remove middle long-chain fat acetoacetic ester the oil phase obtained in step (5), collect molecular distillation heavy phase, be i.e. obtained phosphatidylserine.
In lecithin materials described in step (1), total phospholipids content is 40% ~ 80%; Middle long-chain fat acetoacetic ester described in step (2) is one or more the mixture in ethyl octylate, fish oil ethyl ester, ethyl oleate or DHA-EE; The vigor of step (3) described Phospholipase D is 80 ~ 100U/mg.
The weight ratio of described lecithin materials and middle long-chain fat acetoacetic ester is 1:0.5 ~ 10, the add-on of described Phospholipase D is 1% ~ 10% of lecithin materials weight, the add-on of described Serine is 10% ~ 300% of lecithin materials weight, and the add-on of described damping fluid is 200% ~ 2000% of lecithin materials weight.
In described step (4), the temperature of reaction of heating, stirring reaction is 20 ~ 60 DEG C, and the reaction times is 4 ~ 24h; The distillation temperature of described step (6) Middle molecule distillation is 60 ~ 120 DEG C, and distillation pressure is 0.1 ~ 20Pa.
A kind of phosphatidylserine be prepared from by above-mentioned arbitrary described method.
Prepare a method for phosphatidylserine, comprise the following steps:
(1) from marine animal, lecithin materials is extracted to obtain;
(2) lecithin materials obtained in step (1) is dissolved in middle long-chain fat acetoacetic ester, obtains solution A;
(3) Phospholipase D, Serine and calcium chloride being dissolved in pH value is in the acetate buffer solution of 4.5 ~ 6.0, obtain solution B ';
(4) solution A obtained in step (2) is added the solution B that obtains in step (3) ' in, obtain mixed solution C ', the mixed solution C ' heating obtained, stirring reaction, obtain reacted solution D ';
(5) the reacted solution D ' employing centrifugal method obtained in step (4) is removed aqueous phase and obtain oil phase;
(6) adopt molecular distillation method to remove middle long-chain fat acetoacetic ester the oil phase obtained in step (5), collect molecular distillation heavy phase, be i.e. obtained phosphatidylserine.
In lecithin materials described in step (1), total phospholipids content is 40% ~ 80%; Middle long-chain fat acetoacetic ester described in step (2) is one or more the mixture in ethyl octylate, fish oil ethyl ester, ethyl oleate or DHA-EE; The vigor of step (3) described Phospholipase D is 80 ~ 100U/mg.
The weight ratio of described lecithin materials and middle long-chain fat acetoacetic ester is 1:0.5 ~ 10, the add-on of described Phospholipase D is 1% ~ 10% of lecithin materials weight, the add-on of described Serine is 10% ~ 300% of lecithin materials weight, and the add-on of described damping fluid is 200% ~ 2000% of lecithin materials weight.
In described step (4), the temperature of reaction of heating, stirring reaction is 20 ~ 60 DEG C, and the reaction times is 4 ~ 24h; The distillation temperature of described step (6) Middle molecule distillation is 60 ~ 120 DEG C, and distillation pressure is 0.1 ~ 20Pa.
A kind of phosphatidylserine be prepared from by above-mentioned arbitrary described method.
The present invention adopts such scheme, first reacts with the Baseexchange of Phospholipase D catalysis marine animal phosphatide and Serine, then through centrifugal and molecular distillation, obtained phosphatidylserine after separation and purification; The liquid liquid biphasic reaction system that in utilization, long-chain fat acetoacetic ester and damping fluid are formed, long-chain fat acetoacetic ester substitutes low, the inflammable and explosive and virulent organic solvent of the boiling point such as ether and ethyl acetate with high boiling point and in edible safety, production security is improve while ensure that reaction efficiency, avoid causing environmental pollution, be suitable for suitability for industrialized production.
Embodiment:
Below in conjunction with embodiment, the invention will be further described, but the present invention is not limited thereto, and the preparation method in embodiment is customary preparation methods, no longer describes in detail.
Prepare a method for phosphatidylserine, comprise the following steps:
(1) from marine animal, lecithin materials is extracted to obtain;
(2) lecithin materials obtained in step (1) is dissolved in middle long-chain fat acetoacetic ester, obtains solution A;
(3) Phospholipase D and Serine being dissolved in pH value is in the acetate buffer solution of 4.5 ~ 6.0, obtains solution B;
(4) solution A obtained in step (2) is added in the solution B obtained in step (3), obtain mixed solution C, the mixed solution C heating obtained, stirring reaction, obtain reacted solution D;
(5) adopt centrifugal method to remove aqueous phase the reacted solution D obtained in step (4) and obtain oil phase;
(6) adopt molecular distillation method to remove middle long-chain fat acetoacetic ester the oil phase obtained in step (5), collect molecular distillation heavy phase, be i.e. obtained phosphatidylserine.
In lecithin materials described in step (1), total phospholipids content is 40% ~ 80%; Middle long-chain fat acetoacetic ester described in step (2) is one or more the mixture in ethyl octylate, fish oil ethyl ester, ethyl oleate or DHA-EE; The vigor of step (3) described Phospholipase D is 80 ~ 100U/mg.
The weight ratio of described lecithin materials and middle long-chain fat acetoacetic ester is 1:0.5 ~ 10, the add-on of described Phospholipase D is 1% ~ 10% of lecithin materials weight, the add-on of described Serine is 10% ~ 300% of lecithin materials weight, and the add-on of described damping fluid is 200% ~ 2000% of lecithin materials weight.
In described step (4), the temperature of reaction of heating, stirring reaction is 20 ~ 60 DEG C, and the reaction times is 4 ~ 24h; The distillation temperature of described step (6) Middle molecule distillation is 60 ~ 120 DEG C, and distillation pressure is 0.1 ~ 20Pa.
A kind of phosphatidylserine be prepared from by above-mentioned arbitrary described method.
Prepare a method for phosphatidylserine, comprise the following steps:
(1) from marine animal, lecithin materials is extracted to obtain;
(2) lecithin materials obtained in step (1) is dissolved in middle long-chain fat acetoacetic ester, obtains solution A;
(3) Phospholipase D, Serine and calcium chloride being dissolved in pH value is in the acetate buffer solution of 4.5 ~ 6.0, obtain solution B ';
(4) solution A obtained in step (2) is added the solution B that obtains in step (3) ' in, obtain mixed solution C ', the mixed solution C ' heating obtained, stirring reaction, obtain reacted solution D ';
(5) the reacted solution D ' employing centrifugal method obtained in step (4) is removed aqueous phase and obtain oil phase;
(6) adopt molecular distillation method to remove middle long-chain fat acetoacetic ester the oil phase obtained in step (5), collect molecular distillation heavy phase, be i.e. obtained phosphatidylserine.
In lecithin materials described in step (1), total phospholipids content is 40% ~ 80%; Middle long-chain fat acetoacetic ester described in step (2) is one or more the mixture in ethyl octylate, fish oil ethyl ester, ethyl oleate or DHA-EE; The vigor of step (3) described Phospholipase D is 80 ~ 100U/mg.
The weight ratio of described lecithin materials and middle long-chain fat acetoacetic ester is 1:0.5 ~ 10, the add-on of described Phospholipase D is 1% ~ 10% of lecithin materials weight, the add-on of described Serine is 10% ~ 300% of lecithin materials weight, and the add-on of described damping fluid is 200% ~ 2000% of lecithin materials weight.
In described step (4), the temperature of reaction of heating, stirring reaction is 20 ~ 60 DEG C, and the reaction times is 4 ~ 24h; The distillation temperature of described step (6) Middle molecule distillation is 60 ~ 120 DEG C, and distillation pressure is 0.1 ~ 20Pa.
A kind of phosphatidylserine be prepared from by above-mentioned arbitrary described method.
Embodiment 1:
Extract from marine animal SHISHAMO ovum total phospholipids content be 40% 20g lecithin materials be dissolved in 40g ethyl octylate, obtain solution A; Be that 80U/mg powder lecithin enzyme D, 30gL-Serine is dissolved in 40g, pH value is in the acetate buffer solution of 5.6 by 0.2g vigor, obtain solution B; The solution A obtained is added in the solution B obtained, obtains mixed solution C, the mixed solution C obtained is heated to 20 DEG C of stirring reaction 24h, obtains reacted solution D; Centrifugal method is adopted to isolate oil phase and aqueous phase solution D after reaction terminates, middle long-chain fat acetoacetic ester is removed by all or part of for the oil phase part obtained employing molecular distillation method, distillation temperature in molecular distillation is 60 DEG C, distillation pressure 0.1 ~ 1.0Pa, inlet amount is 2.5g/min, knifing rotating speed is 150rpm, collect molecular distillation heavy phase component i.e. obtained phosphatidylserine, detecting total phospholipids content in obtained phosphatidylserine is 40.2%, and the content of phosphatidylserine in phosphatide is 85.3%.
Embodiment 2:
Extract from marine animal catfish ovum total phospholipids content be 70% 20g lecithin materials be dissolved in 20g fish oil ethyl ester (DHA content is 17.96%), obtain solution A; Be that 85U/mg powder lecithin enzyme D, 35gL-Serine is dissolved in 120g, pH value is in the acetate buffer solution of 6.0 by 0.3g vigor, obtain solution B; The solution A obtained is added in the solution B obtained, obtains mixed solution C, the mixed solution C obtained is heated to 30 DEG C of stirring reaction 20h, obtains reacted solution D; Centrifugal method is adopted to isolate oil phase and aqueous phase solution D after reaction terminates, middle long-chain fat acetoacetic ester is removed by all or part of for the oil phase part obtained employing molecular distillation method, distillation temperature in molecular distillation is 70 DEG C, distillation pressure 1 ~ 5Pa, inlet amount is 2.5g/min, knifing rotating speed is 150rpm, collect molecular distillation heavy phase component i.e. obtained phosphatidylserine, detecting total phospholipids content in obtained phosphatidylserine is 51%, the content of phosphatidylserine in phosphatide is 84.9%, and in the fish oil ethyl ester do not steamed, DHA content is 34.64%.
Embodiment 3:
Extract from marine animal squid ovum total phospholipids content be 50% 2g lecithin materials be dissolved in 20g ethyl oleate, obtain solution A; Be that 95U/mg powder lecithin enzyme D, 2gL-Serine is dissolved in 300g, pH value is in the acetate buffer solution of 4.5 by 1g vigor, obtain solution B; The solution A obtained is added in the solution B obtained, obtains mixed solution C, the mixed solution C obtained is heated to 50 DEG C of stirring reaction 8h, obtains reacted solution D; Centrifugal method is adopted to isolate oil phase and aqueous phase solution D after reaction terminates, middle long-chain fat acetoacetic ester is removed by all or part of for the oil phase part obtained employing molecular distillation method, distillation temperature in molecular distillation is 90 DEG C, distillation pressure 5 ~ 15Pa, inlet amount is 2.5g/min, knifing rotating speed is 150rpm, collect molecular distillation heavy phase component i.e. obtained phosphatidylserine, detecting total phospholipids content in obtained phosphatidylserine is 55%, and the content of phosphatidylserine in phosphatide is 86.5%.
Embodiment 4:
Extract from marine animal flying fish ovum total phospholipids content be 60% 20g lecithin materials be dissolved in 10gDHA ethyl ester, obtain solution A; Be that the calcium chloride that 100U/mg powder lecithin enzyme D, 60gL-Serine and 0.2g can improve Phospholipase D stability is dissolved in 400g, pH value is in the acetate buffer solution of 5.0 by 2g vigor, obtain solution B '; The solution A obtained is added the solution B that obtains ' in, obtain mixed solution C ', the mixed solution C obtained ' be heated to 60 DEG C of stirring reaction 4h, obtain reacted solution D '; After reaction terminates, solution D ' employing centrifugal method is isolated oil phase and aqueous phase, middle long-chain fat acetoacetic ester is removed by all or part of for the oil phase part obtained employing molecular distillation method, distillation temperature in molecular distillation is 120 DEG C, distillation pressure 15 ~ 20Pa, inlet amount is 2.5g/min, knifing rotating speed is 150rpm, collect molecular distillation heavy phase component i.e. obtained phosphatidylserine, detecting total phospholipids content in obtained phosphatidylserine is 58%, and the content of phosphatidylserine in phosphatide is 87.2%.
Adopting existing enzyme transforming process ether to make solvent, from SHISHAMO ovum, prepare the experiment of phosphatidylserine as follows:
Take total phospholipids content in SHISHAMO ovum be 41% 20g lecithin materials be dissolved in 40g ether, it is 80U/mg powder lecithin enzyme D, 30gL-Serine and 0.2g calcium chloride is dissolved in 40g, pH value is in the acetate buffer solution of 5.6 by 0.2g vigor, then by the mixing solutions of lecithin materials and ethyl octylate and the acetate buffer solution mix and blend being dissolved with Phospholipase D, and mixed solution is heated to 20 DEG C of reaction 24h; By mixed solution layering after reaction terminates, collect upper strata ether and volatilize solvent composition i.e. obtained phosphatidylserine mutually, detecting total phospholipids content in obtained phosphatidylserine is 41.3%, and the content of phosphatidylserine in phosphatide is 86.1%.
Adopting existing enzyme transforming process ethyl acetate to make solvent, from SHISHAMO ovum, prepare the experiment of phosphatidylserine as follows:
Take total phospholipids content in SHISHAMO ovum be 41% 20g lecithin materials be dissolved in 40g ethyl acetate, it is 80U/mg powder lecithin enzyme D, 30gL-Serine and 0.2g calcium chloride is dissolved in 40g, pH value is in the acetate buffer solution of 5.6 by 0.2g vigor, then by the mixing solutions of lecithin materials and ethyl octylate and the acetate buffer solution mix and blend being dissolved with Phospholipase D, and mixed solution is heated to 20 DEG C of reaction 24h; By mixed solution layering after reaction terminates, collect upper strata ethyl acetate and volatilize solvent composition i.e. obtained phosphatidylserine mutually, detecting total phospholipids content in obtained phosphatidylserine is 40.5%, and the content of phosphatidylserine in phosphatide is 85.8%.
Sample detection analysis condition is as follows:
The first method that the mensuration of total phospholipids content adopts GB/T5537-2008 to specify measures.
The measuring method of lipid acid composition is as follows:
By sample dissolution in chloroformic solution, point sample is on thin layer silica gel precoated plate, in sherwood oil-ether-acetic acid mixed solution (85:15:1, V/V/V) launch in, phosphatide, triglyceride level and ethyl ester band is scraped after iodine vapor develops the color slightly, adopt gc analysis after esterification respectively, utilize area normalization method to calculate the relative content of each lipid acid.
GC conditions is: gas chromatograph used is be furnished with the Agilent6890 gas chromatograph that injection port is shunting/Splitless injecting samples mouth, and injector temperature is 250 DEG C, and splitting ratio is 20:1; Analytical column selects HP-INNOWax quartz capillary column (30m × 0.32mm × 0.25 μm), adopt gradient increased temperature, initial temperature is 170 DEG C, be warming up to 210 DEG C with 3 DEG C/min and keep 30min, its carrier gas is high pure nitrogen (99.999%), and flow rate of carrier gas is 1mL/min; Detector is flame ionization detector (FID), detector temperature 280 DEG C, and the flow velocity of hydrogen, air and make-up gas (high pure nitrogen) is respectively 40mL/min, 450mL/min and 40mL/min.
The measuring method of phosphatide composition is as follows:
Sample is collecting precipitation part after cold acetone washing, be dissolved in chloroform-methanol mixed solution (4:1, V/V) after volatilizing solvent, carry out efficient liquid phase chromatographic analysis, automatic integration obtains the peak area of each phosphatide composition, calculates the content of each phospholipid composition according to typical curve.
Efficient liquid phase chromatographic analysis is: adopt 1100 type liquid chromatographs, detector is the 2000ELSD type light scattering detector of Alltech company of the U.S., analytical column is Rx-SIL type normal phase silica gel chromatography analytical column (250mm × 4.6mmi.d., 5 μm); Moving phase adopts gradient elution, and mobile phase A is chloroform-methanol-ammoniacal liquor (80:19.5:0.5, V/V/V), Mobile phase B is chloroform-methanol-water-ammonia water (60:34:0.5:5.5, V/V/V/V), flow velocity is 1.0ml/min, and gradient elution program is as shown in the table:
It has been generally acknowledged that, in liquid liquid biphasic reaction system be in the research of catalyst preparing phosphatidylserine with Phospholipase D, the solvent with two substituted oxy group has activation to Phospholipase D, therefore higher reaction efficiency can be obtained by the reaction process of ether or ethyl acetate, but these two kinds of solvent boiling points are low, inflammable and explosive, and have certain toxicity, in actual industrialization is produced, there is safety issue.
And the middle long-chain fat acetoacetic ester that carbonatoms is greater than 10 possesses two substituted oxy group, can add with Phospholipase D be catalyzer liquid liquid biphasic reaction system in for suitability for industrialized production.The boiling point of middle long-chain fat acetoacetic ester is high, not volatile, and possesses certain edible safety, and if the boiling point that can be used as the ethyl octylate of foodstuff additive is 206 DEG C, the fish oil ethyl ester containing EPA and DHA can be used as the production of healthy food material for soft capsule.After reaction terminates, by long-chain fat acetoacetic ester in all or part of removing of molecular distillation technique.Phosphatide non-refractory, but research finds, and under the condition of high vacuum, the existence of neutral grease (as fatty-acid ethyl ester, triglyceride level etc.) has provide protection to the phosphatide under high temperature.
Elder generation of the present invention reacts with the Baseexchange of Phospholipase D catalysis marine animal phosphatide and Serine, then through centrifugal and molecular distillation, obtained phosphatidylserine after separation and purification; The liquid liquid biphasic reaction system that in utilization, long-chain fat acetoacetic ester and damping fluid are formed, long-chain fat acetoacetic ester substitutes low, the inflammable and explosive and virulent organic solvent of the boiling point such as ether and ethyl acetate with high boiling point and in edible safety, production security is improve while ensure that reaction efficiency, avoid causing environmental pollution, be suitable for suitability for industrialized production.
The above is only preferred embodiment of the present invention, and be not restriction the present invention being made to other form, any those skilled in the art may utilize the technology contents of above-mentioned announcement to be changed or be modified as the Equivalent embodiments of equivalent variations.In every case be do not depart from technical solution of the present invention content, any simple modification, equivalent variations and the remodeling done above embodiment according to technical spirit of the present invention, still belong to the protection domain of technical solution of the present invention.
The present invention does not describe part in detail, is the known technology of those skilled in the art of the present technique.
Claims (10)
1. prepare a method for phosphatidylserine, it is characterized in that: comprise the following steps:
(1) from marine animal, lecithin materials is extracted to obtain;
(2) lecithin materials obtained in step (1) is dissolved in middle long-chain fat acetoacetic ester, obtains solution A;
(3) Phospholipase D and Serine being dissolved in pH value is in the acetate buffer solution of 4.5 ~ 6.0, obtains solution B;
(4) solution A obtained in step (2) is added in the solution B obtained in step (3), obtain mixed solution C, the mixed solution C heating obtained, stirring reaction, obtain reacted solution D;
(5) adopt centrifugal method to remove aqueous phase the reacted solution D obtained in step (4) and obtain oil phase;
(6) adopt molecular distillation method to remove middle long-chain fat acetoacetic ester the oil phase obtained in step (5), collect molecular distillation heavy phase, be i.e. obtained phosphatidylserine.
2. a kind of method preparing phosphatidylserine according to claim 1, is characterized in that: in the lecithin materials described in step (1), total phospholipids content is 40% ~ 80%; Middle long-chain fat acetoacetic ester described in step (2) is one or more the mixture in ethyl octylate, fish oil ethyl ester, ethyl oleate or DHA-EE; The vigor of step (3) described Phospholipase D is 80 ~ 100U/mg.
3. a kind of method preparing phosphatidylserine according to claim 1, it is characterized in that: the weight ratio of described lecithin materials and middle long-chain fat acetoacetic ester is 1:0.5 ~ 10, the add-on of described Phospholipase D is 1% ~ 10% of lecithin materials weight, the add-on of described Serine is 10% ~ 300% of lecithin materials weight, and the add-on of described damping fluid is 200% ~ 2000% of lecithin materials weight.
4. a kind of method preparing phosphatidylserine according to claim 1, is characterized in that: in described step (4), the temperature of reaction of heating, stirring reaction is 20 ~ 60 DEG C, and the reaction times is 4 ~ 24h; The distillation temperature of described step (6) Middle molecule distillation is 60 ~ 120 DEG C, and distillation pressure is 0.1 ~ 20Pa.
5. one kind as arbitrary in Claims 1-4 as described in the phosphatidylserine that is prepared from of method.
6. prepare a method for phosphatidylserine, it is characterized in that: comprise the following steps:
(1) from marine animal, lecithin materials is extracted to obtain;
(2) lecithin materials obtained in step (1) is dissolved in middle long-chain fat acetoacetic ester, obtains solution A;
(3) Phospholipase D, Serine and calcium chloride being dissolved in pH value is in the acetate buffer solution of 4.5 ~ 6.0, obtain solution B ';
(4) solution A obtained in step (2) is added the solution B that obtains in step (3) ' in, obtain mixed solution C ', the mixed solution C ' heating obtained, stirring reaction, obtain reacted solution D ';
(5) the reacted solution D ' employing centrifugal method obtained in step (4) is removed aqueous phase and obtain oil phase;
(6) adopt molecular distillation method to remove middle long-chain fat acetoacetic ester the oil phase obtained in step (5), collect molecular distillation heavy phase, be i.e. obtained phosphatidylserine.
7. a kind of method preparing phosphatidylserine according to claim 6, is characterized in that: in the lecithin materials described in step (1), total phospholipids content is 40% ~ 80%; Middle long-chain fat acetoacetic ester described in step (2) is one or more the mixture in ethyl octylate, fish oil ethyl ester, ethyl oleate or DHA-EE; The vigor of step (3) described Phospholipase D is 80 ~ 100U/mg.
8. a kind of method preparing phosphatidylserine according to claim 6, it is characterized in that: the weight ratio of described lecithin materials and middle long-chain fat acetoacetic ester is 1:0.5 ~ 10, the add-on of described Phospholipase D is 1% ~ 10% of lecithin materials weight, the add-on of described Serine is 10% ~ 300% of lecithin materials weight, and the add-on of described damping fluid is 200% ~ 2000% of lecithin materials weight.
9. a kind of method preparing phosphatidylserine according to claim 6, is characterized in that: in described step (4), the temperature of reaction of heating, stirring reaction is 20 ~ 60 DEG C, and the reaction times is 4 ~ 24h; The distillation temperature of described step (6) Middle molecule distillation is 60 ~ 120 DEG C, and distillation pressure is 0.1 ~ 20Pa.
10. one kind as arbitrary in claim 6 to 9 as described in the phosphatidylserine that is prepared from of method.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510566411.1A CN105177073B (en) | 2015-09-08 | 2015-09-08 | A kind of method preparing Phosphatidylserine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510566411.1A CN105177073B (en) | 2015-09-08 | 2015-09-08 | A kind of method preparing Phosphatidylserine |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105177073A true CN105177073A (en) | 2015-12-23 |
| CN105177073B CN105177073B (en) | 2016-06-08 |
Family
ID=54899499
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510566411.1A Active CN105177073B (en) | 2015-09-08 | 2015-09-08 | A kind of method preparing Phosphatidylserine |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105177073B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107334008A (en) * | 2017-06-14 | 2017-11-10 | 芜湖福民生物药业股份有限公司 | Fruit beverage containing phosphatidylserine and preparation method thereof |
| CN107688073A (en) * | 2017-11-02 | 2018-02-13 | 威海百合生物技术股份有限公司 | A kind of detection method of phosphatidylserine content |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103131737A (en) * | 2011-11-26 | 2013-06-05 | 江南大学 | Method of preparing phosphatidylserine in enzymic mode |
| CN103509047A (en) * | 2012-06-22 | 2014-01-15 | 上海傲硕生物科技有限公司 | Extraction process of phosphatidylcholine with antarctic krill as source and preparation method of phosphatidylserine |
-
2015
- 2015-09-08 CN CN201510566411.1A patent/CN105177073B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103131737A (en) * | 2011-11-26 | 2013-06-05 | 江南大学 | Method of preparing phosphatidylserine in enzymic mode |
| CN103509047A (en) * | 2012-06-22 | 2014-01-15 | 上海傲硕生物科技有限公司 | Extraction process of phosphatidylcholine with antarctic krill as source and preparation method of phosphatidylserine |
Non-Patent Citations (2)
| Title |
|---|
| 胡飞，等: "生物酶法制备磷脂酰丝氨酸的研究进展", 《中国油脂》, vol. 37, no. 6, 31 December 2012 (2012-12-31) * |
| 路福平，等: "双液相体系中磷脂酶D转酯合成磷脂酰丝氨酸的研究", 《2007年全国发酵行业高峰论坛》, 31 December 2007 (2007-12-31) * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107334008A (en) * | 2017-06-14 | 2017-11-10 | 芜湖福民生物药业股份有限公司 | Fruit beverage containing phosphatidylserine and preparation method thereof |
| CN107688073A (en) * | 2017-11-02 | 2018-02-13 | 威海百合生物技术股份有限公司 | A kind of detection method of phosphatidylserine content |
| CN107688073B (en) * | 2017-11-02 | 2020-12-04 | 威海百合生物技术股份有限公司 | Method for detecting content of phosphatidylserine |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105177073B (en) | 2016-06-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Kim et al. | Ionic liquid-mediated extraction of lipids from algal biomass | |
| Billakanti et al. | Enzyme-assisted extraction of fucoxanthin and lipids containing polyunsaturated fatty acids from Undaria pinnatifida using dimethyl ether and ethanol | |
| Balasubramanian et al. | Factors affecting cellular lipid extraction from marine microalgae | |
| Isaksson et al. | Lipid residue analyses of Early Neolithic funnel-beaker pottery from Skogsmossen, eastern Central Sweden, and the earliest evidence of dairying in Sweden | |
| CN104651422B (en) | A kind of method that triglyceride type DHA and EPA are extracted from deep-sea fish | |
| Zhang et al. | Why solid-state fermentation is more advantageous over submerged fermentation for converting high concentration of glycerol into Monacolin K by Monascus purpureus 9901: A mechanistic study | |
| Papakosta et al. | Early pottery use across the Baltic–a comparative lipid residue study on Ertebølle and Narva ceramics from coastal hunter-gatherer sites in southern Scandinavia, northern Germany and Estonia | |
| CN105821088A (en) | Method for preparing glyceride rich in EPA and DHA through enzyme catalysis | |
| Cheong et al. | Extraction and enrichment of n-3 polyunsaturated fatty acids and ethyl esters through reversible π–π complexation with aromatic rings containing ionic liquids | |
| CN109468349A (en) | A kind of method that enzyme process prepares 1- oleic acid -2- palmitinic acid -3- linoleic acid triglyceride | |
| Prommuak et al. | Simultaneous production of biodiesel and free lutein from Chlorella vulgaris | |
| Michalak et al. | Supercritical algal extracts: a source of biologically active compounds from nature | |
| CN103320217A (en) | Method for extracting krill oil rich in phospholipid from euphausia superba | |
| CN107058413A (en) | The method of Long carbon chain triglycerides in prepared by a kind of utilization camphor tree seeds oil and soybean oil | |
| CN105016956A (en) | Squalene extracting method | |
| CN103952448B (en) | A kind of method utilizing enzyme-chemically method directional preparation OPO | |
| CN103254993A (en) | Rice bran crude oil separation method | |
| CN105177073A (en) | Method of preparing phosphatidylserine | |
| CN102630943A (en) | Spirulina gamma-linolenic acid extractive and preparation method thereof | |
| CN102838684B (en) | Separating and purifying process of isochrysis galbana exopolysaccharide | |
| He et al. | Separation of saturated fatty acids from docosahexaenoic acid‐rich algal oil by enzymatic ethanolysis in tandem with molecular distillation | |
| CN105296142B (en) | Method for rapidly extracting and separating lipid components in peony seeds | |
| CN103509047B (en) | The extraction process of the phosphatidylcholine of a kind of antarctic krill and the preparation method of Phosphatidylserine | |
| Takatsuki et al. | Liquid chromatographic determination of free and added niacin and niacinamide in beef and pork | |
| CN1451729A (en) | Method for extracting yolk oil by enzyme hydrolysis-solvent extraction |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB03 | Change of inventor or designer information |
Inventor after: Sun Zhaomin Inventor after: Cai Shengli Inventor after: Cai Aiying Inventor after: Huang Shasha Inventor after: Li Wenping Inventor before: Cai Shengli Inventor before: Cai Aiying Inventor before: Sun Zhaomin Inventor before: Huang Shasha Inventor before: Li Wenping |
|
| COR | Change of bibliographic data | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: A method for preparing phosphatidylserine Effective date of registration: 20220621 Granted publication date: 20160608 Pledgee: Weihai commercial bank Limited by Share Ltd. revitalization branch Pledgor: WEIHAI BOYU FOOD Co.,Ltd. Registration number: Y2022980008452 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20230629 Granted publication date: 20160608 Pledgee: Weihai commercial bank Limited by Share Ltd. revitalization branch Pledgor: WEIHAI BOYU FOOD Co.,Ltd. Registration number: Y2022980008452 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: A method for preparing phosphatidylserine Effective date of registration: 20230703 Granted publication date: 20160608 Pledgee: Weihai commercial bank Limited by Share Ltd. revitalization branch Pledgor: WEIHAI BOYU FOOD Co.,Ltd. Registration number: Y2023980047050 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Granted publication date: 20160608 Pledgee: Weihai commercial bank Limited by Share Ltd. revitalization branch Pledgor: WEIHAI BOYU FOOD Co.,Ltd. Registration number: Y2023980047050 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: A method for preparing phosphatidylserine Granted publication date: 20160608 Pledgee: Weihai commercial bank Limited by Share Ltd. revitalization branch Pledgor: WEIHAI BOYU FOOD Co.,Ltd. Registration number: Y2024980027930 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right |