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Process for preparing choline glycerophosphatide (GPC) with non-aqueous phase enzymatic method

A technology of glycerophosphocholine and enzymatic preparation, which is applied in the direction of fermentation, can solve the problems of expensive reagents, high environmental pollution, difficult control of reaction conditions, etc., achieve mild conditions, high yield, and change the production process.

Inactive Publication Date: 2012-10-31
曹明成 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, many studies have been done on the preparation of GPC by chemical method, and the yield is relatively high, but there are some disadvantages in the preparation method of chemical method, such as difficult to control the reaction conditions, expensive reagents, and large environmental pollution.

Method used

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  • Process for preparing choline glycerophosphatide (GPC) with non-aqueous phase enzymatic method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Take 5g of soybean lecithin (PC50) and ultrasonically dissolve it in 41ml of heptane solvent, then add dropwise the enzyme solution containing 2.5ml of phospholipase A1 and 10ml of acetic acid / sodium acetate buffer solution of pH5.0, and place it at 40 Stir and react at constant temperature for 4 hours at ℃, take out the reaction solution, and then take an extraction agent equal to the volume of the reaction solution, that is, CH 3 OH and distilled water mixed solvent for extraction (CH in the extractant 3 OH and distilled water volume ratio 1:1.5), take the lower layer of water, concentrate and dry to get the GPC product; the crude GPC product obtained after the reaction liquid is extracted and purified can be qualitatively analyzed and detected by TLC method: the selected developing agent is: dichloro Methane:methanol:triethylamine=35:60:5, after the sample was spotted and developed, its Rf value was calculated to be 0.28 by comparing it with the GPC standard. At the...

Embodiment 2

[0025] Take 5g of egg yolk lecithin (PC70) and ultrasonically dissolve it in 41ml of heptane solvent, then add dropwise the enzyme solution containing 2.5ml of phospholipase A1 and 5ml of acetic acid / sodium acetate buffer solution with pH4.0, and place it at 40 Stir and react at constant temperature for 4 hours at ℃, take out the reaction solution, and then take an extraction agent equal to the volume of the reaction solution, that is, CH 3 OH and distilled water mixed solvent for extraction (CH in the extractant 3 OH and distilled water volume ratio 1:1.5), after extraction, the water phase is concentrated and dried to obtain the GPC product; the crude GPC product obtained after the reaction liquid is extracted and purified can be qualitatively analyzed and detected by TLC. First of all, choose a suitable developing agent, so that the GPC and impurities in the sample can be effectively separated on the thin-layer plate. After exploration, the developing agent is finally deter...

Embodiment 3

[0028] Take 5g of soybean lecithin (PC70) and ultrasonically dissolve it in 41ml of heptane solvent, then add dropwise the enzyme solution containing 2.5ml of phospholipase A1 and 5ml of acetic acid / sodium acetate buffer solution of pH4.0, and place it at 40 Stir and react at a constant temperature for 4 hours at ℃, take out the reaction solution, and then take an extraction agent that is 3 times the volume of the reaction solution, that is, CH 3 OH and distilled water mixed solvent for extraction (CH in the extractant 3 OH and distilled water volume ratio 1:1.5), after extraction, the water phase is concentrated and dried to obtain the GPC product; the crude GPC product obtained after the reaction liquid is extracted and purified can be qualitatively analyzed and detected by TLC. First of all, choose a suitable developing agent, so that the GPC and impurities in the sample can be effectively separated on the thin-layer plate. After exploration, the developing agent is finally...

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Abstract

The invention discloses a process for preparing choline glycerophosphatide (GPC) with a non-aqueous phase enzymatic method. The process comprises the steps that: lecithin is dissolved in a heptane solvent; an enzyme-containing solution containing phospholipase A1 and an acetic acid / acetate buffer solution is dropped into the solution; the mixture is subjected to a constant-temperature reaction under a temperature of 30-60 DEG C; a reaction liquid is extracted; and an aqueous phase is concentrated and is dried by baking, so that the GPC product is obtained. In the adopted lecithin, the content of phosphatidyl choline is no lower than 50%. An adopted extractant is a mixed solvent of CH3OH and water. The invention provides the process for preparing choline glycerophosphatide (GPC) with the non-aqueous phase enzymatic method. The process adopts an appropriate reaction system, and assists in preparing GPC products with high yield and high purity. Therefore, domestic existing GPC production processes are fundamentally changed, and GPC products with excellent quality can be prepared highly efficiently with mild conditions.

Description

technical field [0001] The invention belongs to the technical field of preparing glycerophosphorylcholine by a biological method, and specifically relates to a method for preparing GPC by using a phospholipase capable of hydrolyzing two fatty acid chains in lecithin to catalyze the breaking of ester chains in a non-aqueous phase system. [0002] Background technique [0003] Glycerophosphorylcholine (GPC) is one of the products of phospholipid metabolism in the body. In vivo, the most important physiological function of GPC is to cross the blood-brain barrier and provide the necessary choline for the biosynthesis of acetylcholine and phosphatidylcholine (PC). Acetylcholine is an important neurotransmitter in the central nervous system, which helps the brain to complete learning, memory and cognitive activities, and can control light sleep and muscle activity, and can even repair the partially damaged cognition of early-stage dementia patients ability. Increase the synthes...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/00
Inventor 张洪斌毛慧芳曹明成
Owner 曹明成
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