CN101157946A - Technique for preparing phosphatidyl serine rich in highly-unsaturated fatty acid - Google Patents
Technique for preparing phosphatidyl serine rich in highly-unsaturated fatty acid Download PDFInfo
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- CN101157946A CN101157946A CNA2007101138080A CN200710113808A CN101157946A CN 101157946 A CN101157946 A CN 101157946A CN A2007101138080 A CNA2007101138080 A CN A2007101138080A CN 200710113808 A CN200710113808 A CN 200710113808A CN 101157946 A CN101157946 A CN 101157946A
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Abstract
The invention relates to a technique for preparing diacylglyceryl-phosphorylserine rich in unsaturated acid, which is characterized in that a reactor is preheated to 30 to 60 DEG C, squid lecithin and L-serine at the mass ratio of 1:1-15 is added, phosphatidase D2-15U/g is also added as reaction substrate, gas R134a is pumped to lead the pressure of the reactor to reach 4-6MPa, the pressure is reduced after agitating reaction for 2 to 8 hours, R134a is recycled after the reaction mixture comes into a separation pot which is communicated with the reactor under the temperature of 30 to 60 DEG C, reaction mixture is collected from the a sample receiving port of the separation pot, at last, the reaction mixture is washed by using acetone and centrifuged for 5 to 10 min to separate the solid phase for further purification. The diacylglyceryl-phosphorylserine prepared by the invention rich in high unsaturated fatty acid as the medium in catalytic reaction of phosphatidase D has the advantages of simple technique, low equipment input, short reaction time, high content of unsaturated fatty acid and high yield and the neglecting of organic solvent. Furthermore, R134a can be completely recycled after reaction, which is beneficial to the separation of products and repeated use of enzyme.
Description
Technical field
The present invention relates to a kind of preparation and be rich in the technology of the phosphatidylserine of polyunsaturated fatty acid.
Background technology
N-3 polyunsaturated fatty acid in the fish oil (n-3 PUFA); especially timnodonic acid (EPA) and docosahexenoic acid (DHA) have that prevention and treatment cardiovascular disorder, anti-inflammatory are anticancer, strengthening immunity, help physiologically actives such as brain development and vision protection.Phosphatidylserine (PS) is the important composition composition of mammalian cell membrane, as widespread uses in medicine, makeup, foodstuffs industry such as liposome, emulsifying agents.Phosphatidylserine also has the activity of improving the memory function of suffering from old dysmnesia and AlzheimerShi disease patient.The processing step that extracts phosphatidylserine from natural product is numerous and diverse, and yield is low, and valuable product has limited the further widespread use of phosphatidylserine.In recent years, utilize Phospholipase D catalyzing acyl shift reaction, from phosphatidylcholine (PC) the preparation PS that raw material is easy to get, existing bibliographical information.From sleeve-fish-processing waste---the PC that obtains the squid ovum, EPA and DHA content were very high during its lipid acid was formed, with it is raw material, the PS of the high n-3 high unsaturated fatty acid by enzymatic reaction preparation can have the biological function of n-3 polyunsaturated fatty acid and PS concurrently, has realized the higher value application of waste.Enzymic catalytic reaction in the past mainly is to carry out in traditional organic solvent (normal hexane etc.), long reaction time, and the product separating step is loaded down with trivial details, and product yield is low.
Summary of the invention
The purpose of this invention is to provide a kind of technology that is rich in the high unsaturated fatty acid phosphatidylserine that in subcritical R134a system, prepares, to overcome the above-mentioned shortcoming of existing technology.
The technology of the phosphatidylserine of polyunsaturated fatty acid is rich in a kind of preparation, it is characterized in that earlier reactor being heated to 30 ℃-60 ℃, in reactor, add squid Yelkin TTS and L-Serine, both mass ratioes are 1: 1-15, and adding Phospholipase D 2-15U/g reaction substrate, closed reactor afterwards, pump into R134a gas and make reactor pressure reach 4-6MPa, after stirring reaction 2-8 hour, decompression, the temperature that reaction mixture is entered be communicated with reactor is 30-60 ℃ a separating still, reclaim R134a, collect reaction mixture from the sample mouth that connects of separating still again, use the washing with acetone reaction mixture at last, tell solid phase in centrifugal 5-10 minute, carry out purifying again.
The present invention utilizes the medium of subcritical R134a as the Phospholipase D catalyzed reaction, the phosphatidylserine of high unsaturated fatty acid is rich in preparation, and technology is simple, and equipment drops into low, speed of response is fast, time is short, makes the content height of the unsaturated fatty acids in the product, product yield height, and reaction need not organic solvent, finishing back R134a in reaction can reclaim fully, is beneficial to the separation of product and the recycling of enzyme, realizes green reaction process.
Embodiment
Embodiment one
Reactor is heated to 40 ℃, takes by weighing squid Yelkin TTS 75mg, place reactor; Take by weighing Calcium Chloride Powder Anhydrous (being used to keep the stability of enzyme) 0.1g, solid L-Serine 0.85g is dissolved in the phosphate buffered saline buffer of 2.4mlpH5.5, adds in the reactor; Precision takes by weighing Phospholipase D 9.6U, adds in the reactor; After reactor is airtight, opens and be equipped with 1,1,1, the storage tank of 2-Tetrafluoroethane (R134a) makes R134a gas enter reactor by the ram pump pressurization after condenser liquefaction.After reactor pressure reached 6MPa, R134a reached subcritical state.The stop plunger pump cuts out storage tank, stirring reaction.React after 3.5 hours, open shut off valve decompression, the temperature that reaction mixture is entered be communicated with reactor is 40 ℃ a separating still.Open compressor and reclaim R134a from separating still; After recovery is finished, collect reaction mixture from the sample mouth that connects of separating still.With 10ml washing with acetone reaction mixture, centrifugal 5 minutes, to tell solid phase, purifying and promptly get the phosphatidylserine 58mg that is rich in polyunsaturated fatty acid, transformation efficiency is 90.6%, the content of EPA, DHA is respectively 12.42%, 32.09%.
Embodiment two
Reactor and separating still are heated to 40 ℃, take by weighing squid Yelkin TTS 300mg, place reactor; Take by weighing Calcium Chloride Powder Anhydrous 0.4g, solid L-Serine 3.5g is dissolved in the phosphate buffered saline buffer of 10mlpH5.5, adds in the reactor; Precision takes by weighing Phospholipase D 38U, adds in the reactor; After system is airtight, open the R134a storage tank, make R134a gas after condenser liquefaction, enter reactor by the ram pump pressurization.After reactor pressure reached 6MPa, the stop plunger pump cut out storage tank, stirring reaction.React after 3.5 hours, open shut off valve decompression, the temperature that reaction mixture is entered be communicated with reactor is 40 ℃ a separating still.Open compressor and reclaim R134a from separating still; After recovery is finished, collect reaction mixture from the sample mouth that connects of separating still.With 20ml washing with acetone reaction mixture, centrifugal 5 minutes, to tell solid phase, purifying and promptly get the phosphatidylserine 260mg that is rich in high unsaturated fatty acid, transformation efficiency is 91.3%, the content of EPA, DHA is 12.39%, 32.13%.
The mass ratio of squid Yelkin TTS of the present invention and L-Serine is 1: 1-15, the amount that every gram reaction substrate adds Phospholipase D is 2-15U, temperature of reaction is 30-60 ℃, reaction pressure is 4-6MPa, reaction times is 3-8 hour, described subcritical one of fluidic pressure, these two decisive processing parameters of temperature of being meant are on the stagnation point, and another parameter is in subcritical state.R134a is a kind of novel refrigerant harmless to atmospheric ozone layer, and its critical temperature is 101.1 ℃; Its emergent pressure is lower than CO
2Emergent pressure, and the dissolving power of subcritical R134a is higher than supercritical CO
2, can guarantee enough dissolving poweies to solute, can reduce disposable apparatus input and working cost greatly again.The viscous modulus of subcritical R134a is less than its supercritical state, and spread coefficient can help reducing the diffusional limitation that substrate contacts with enzyme in the enzymic catalytic reaction more greater than its supercritical state, improves enzymatic kinetics, improves speed of reaction.
Claims (1)
1. technology for preparing the phosphatidylserine that is rich in high unsaturated fatty acid, it is characterized in that earlier reactor being heated to 30 ℃-60 ℃, in reactor, add squid Yelkin TTS and L-Serine, both mass ratioes are 1: 1-15, and adding Phospholipase D 2-15U/g reaction substrate, closed reactor afterwards, pump into R134a gas and make reactor pressure reach 4-6MPa, after stirring reaction 2-8 hour, decompression, the temperature that reaction mixture is entered be communicated with reactor is 30-60 ℃ a separating still, reclaim R134a, collect reaction mixture from the sample mouth that connects of separating still again, use the washing with acetone reaction mixture at last, tell solid phase in centrifugal 5-10 minute, carry out purifying again.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007101138080A CN101157946A (en) | 2007-09-13 | 2007-09-13 | Technique for preparing phosphatidyl serine rich in highly-unsaturated fatty acid |
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| CNA2007101138080A CN101157946A (en) | 2007-09-13 | 2007-09-13 | Technique for preparing phosphatidyl serine rich in highly-unsaturated fatty acid |
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| CN101157946A true CN101157946A (en) | 2008-04-09 |
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| CNA2007101138080A Pending CN101157946A (en) | 2007-09-13 | 2007-09-13 | Technique for preparing phosphatidyl serine rich in highly-unsaturated fatty acid |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012071768A1 (en) * | 2010-12-03 | 2012-06-07 | Zhang Yongzhi | Method for preparing high quality euphausia superba oil with phosphatidylserine enriched of polyunsaturated double bond fatty acyl group |
| CN103966277A (en) * | 2014-05-22 | 2014-08-06 | 南京工业大学 | Method for preparing phosphatidylserine under catalysis of immobilized phospholipase D |
| CN105950584A (en) * | 2016-03-30 | 2016-09-21 | 中国海洋大学 | Phospholipase D and application thereof |
-
2007
- 2007-09-13 CN CNA2007101138080A patent/CN101157946A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012071768A1 (en) * | 2010-12-03 | 2012-06-07 | Zhang Yongzhi | Method for preparing high quality euphausia superba oil with phosphatidylserine enriched of polyunsaturated double bond fatty acyl group |
| CN103966277A (en) * | 2014-05-22 | 2014-08-06 | 南京工业大学 | Method for preparing phosphatidylserine under catalysis of immobilized phospholipase D |
| CN105950584A (en) * | 2016-03-30 | 2016-09-21 | 中国海洋大学 | Phospholipase D and application thereof |
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Open date: 20080409 |