Method for preparing phosphatidylserine abundant in polyunsaturated fatty acid

A technology of unsaturated fatty acid and phosphatidylserine, applied in the field of bioengineering, can solve the problems of high manufacturing cost, difficult industrialization, complex device structure, etc., and achieve the effects of easy large-scale production, few reaction by-products, and simple production process

Inactive Publication Date: 2010-09-01
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These reactions all occur in the organic phase, the process operation is unsafe, and the product has solvent residue, which is harmful to health
Li Zhaojie and others used PLD to catalyze the reaction of squid lecithin (rich in EPA, DHA) and L-serine to prepare PS in the subcritical system (patent CN101157946), but the content of such squid lecithin in nature is limited, and the subcritical fluid is related. The structure of the device is complicated, the manufacturing cost is high, and it is limited to a certain subcritical fluid, so it is difficult to industrialize

Method used

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  • Method for preparing phosphatidylserine abundant in polyunsaturated fatty acid
  • Method for preparing phosphatidylserine abundant in polyunsaturated fatty acid
  • Method for preparing phosphatidylserine abundant in polyunsaturated fatty acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Weigh 0.75g egg yolk lecithin (purity ≥ 99%) and 0.918g gamma-linolenic acid ethyl ester (containing gamma-linolenic acid 72.72%) in a flat-bottomed glass sample bottle with a stopper, fill the bottle with nitrogen, at 50 ° C Stir to make lecithin completely dissolved in ethyl ester, then add phospholipase A by 10wt% 1 ( Ultra), seal the sample bottle, fill the bottle with nitrogen gas again, and stir the reaction at 50°C for 10h. After the reaction is completed, add 1.0mol / L HCl to the sample to terminate the reaction, and then add an equal amount of 1.0mol / L NaOH And, then add 18 mL of 0.2 mol / L sodium acetate buffer solution (pH5.6) containing 3.4 mol / L of L-serine and 4.8 U of Streptomyces phospholipase D to the sample, at 30 ° C, 200 r / min After the reaction was completed for 16 hours, 1.0 mol / L HCl was added to the sample to terminate the reaction. Take a small amount of sample, add an appropriate amount of chloroform solution, then centrifuge the sample at 400...

Embodiment 2

[0045] Weigh 1.25g soybean lecithin (purity ≥ 60%) and 1.05g fish oil ethyl ester (containing EPA9%, DHA42%) in a flat-bottomed glass sample bottle with a stopper, fill the bottle with nitrogen, and stir at 55°C to make the eggs Phospholipids are completely dissolved in ethyl ester, and phospholipase A is added at 20 wt% 1 ( Novo), seal the sample bottle, fill the bottle with nitrogen again, stir and react at 55°C for 10h, after the reaction is completed, add 1.0mol / L HCl to the sample to terminate the reaction, and then add an equivalent amount of 1.0mol / L NaOH And, then add 20 mL of 0.11 mol / L sodium acetate buffer solution (pH5.6) containing 2.0 mol / L of L-serine and 10 U of soybean phospholipase D to the sample, and react at 28 °C and 200 r / min for 18 h , after the reaction was completed, 1.0 mol / L HCl was added to the sample to terminate the reaction. Take a small amount of sample, add an appropriate amount of chloroform solution, then centrifuge the sample at 4000r / mi...

Embodiment 3

[0047] Weigh 0.95g of soybean lecithin (purity ≥ 90%) and 3.60g of fish oil (containing DHA and EPA50%) in a flat-bottomed glass sample bottle with a stopper, fill the bottle with nitrogen, and stir at 50°C to completely dissolve the lecithin In the fish oil, add Novozym 435 at 10wt%, seal the sample bottle, fill the bottle with nitrogen gas again, stir and react at 50°C for 14h, after the reaction is completed, filter to remove the enzyme, and then add 30mL of L containing 2.0mol / L to the sample - 0.22mol / L sodium acetate buffer solution (pH5.6) of serine and 6U of phospholipase D (derived from coryneform bacteria), reacted at 30°C and 200r / min for 24h, after the reaction was completed, add 1.0 mol / L HCl to terminate the reaction. Take a small amount of sample, add an appropriate amount of chloroform solution, then centrifuge the sample at 4000r / min for 10min, take the chloroform layer, analyze the sample, use TLC-GC to detect the fatty acid composition of phospholipids, and ...

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Abstract

The invention relates to a method for preparing phosphatidylserine abundant in polyunsaturated fatty acid and belongs to the technical field of bioengineering. The method is characterized by comprising the following steps of: firstly, catalyzing ester exchange reaction between phosphatide and polyunsaturated fatty acid ester by utilizing one or a mixture of phosphatidase A and lipase to generate phosphatide abundant in polyunsaturated fatty acid; and then catalyzing phosphor-transfer esterification reaction between the phosphatide abundant in the polyunsaturated fatty acid and L-serine by utilizing phosphatidase D to generate the phosphatidylserine abundant in the polyunsaturated fatty acid. The method has the advantages of no discharge of waste water, good product quality, no solvent residue, safe process operation, few reaction byproducts, no waste generation, cost reduction, simple production process and easy realization of scale production because of utilizing two enzymes to perform sub-step catalysis and perform reaction in the same reactor, and completing the reaction process in a non-solvent system. Therefore, the invention provides a good and feasible method for preparing the phosphatidylserine abundant in the polyunsaturated fatty acid.

Description

technical field [0001] The invention belongs to the technical field of bioengineering and relates to a method for preparing phosphatidylserine rich in polyunsaturated fatty acids. Background technique [0002] Phospholipids are an important component of cell membranes and play an important role in maintaining the normal life activities of cells. They are also natural surfactants with physiological effects and are widely used in food, medicine, cosmetics, health products and feed industries. Phospholipids mainly include phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS) and phosphatidic acid (PA). The physiological activity of phospholipids is closely related to the composition of its fatty acids and the properties of polar ends, and some phospholipids with specific structures have high medicinal value. The following formula is the structural formula of phosphoglycerides. [0003] [0004] Glyceryl Phosphate Struct...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/06C12P39/00C12R1/01C12R1/085C12R1/15C12R1/19C12R1/38C12R1/39C12R1/42C12R1/46C12R1/465C12R1/645C12R1/68C12R1/685C12R1/69C12R1/72C12R1/785C12R1/80C12R1/845C12R1/85
Inventor 杨天奎赵紫薇牟英
Owner DALIAN UNIV OF TECH
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