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Bioconversion method of L-hydroxyproline

A biotransformation and hydroxyproline technology, applied in the field of L-hydroxyproline biotransformation, can solve the problems of low optical purity and low yield, and achieve simple process flow, strong stereoselectivity and ratio reasonable effect

Inactive Publication Date: 2017-12-05
绍兴百茵生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method is purely prepared by biological methods, the yield is low, and the optical purity is not high

Method used

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  • Bioconversion method of L-hydroxyproline
  • Bioconversion method of L-hydroxyproline
  • Bioconversion method of L-hydroxyproline

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] L- Biotransformation method of hydroxyproline , the reaction was carried out in a 1L shake flask, and the reaction system was controlled to 300mL, including 20g of L-proline and 10g of α-ketoglutarate as substrates, and 4.2g of proline-4-hydroxylase engineering The whole cell of the bacteria is used as a catalyst, and a surfactant and an additive are added to establish a biotransformation system to carry out a transformation reaction to obtain a transformation solution containing L-hydroxyproline. Among them, the surfactant is composed of 0.011g sorbitan ester, 0.132g Tween 80, and 0.004g triton X-100. The additive is a mixture of 0.048g ferrous sulfate, 0.035g ferrous chloride and 0.014g ferrous nitrate. The temperature of the biotransformation system is controlled at 35° C.; the pH value of the biotransformation system is controlled at 6; the transformation reaction is carried out in a shaking table, and the rotating speed of the shaking table is controlled at 200 ...

Embodiment 2

[0058] L- Biotransformation method of hydroxyproline , the reaction was carried out in a 5L beaker, and the reaction system was controlled to 2L, including 200g of L-proline, 65g of α-ketoglutarate as substrates, and 40g of proline-4-hydroxylase engineering bacteria Cells are used as catalysts, surfactants and additives are added to establish a biotransformation system for transformation reactions, and transformation liquid containing L-hydroxyproline is obtained. Among them, the surfactant is composed of 0.076g sorbitan ester, 0.703g Tween 80, and 0.022g triton X-100. Additive is the mixture of 1g ferrous sulfate, 0.667g ferrous chloride, 0.333g ferrous nitrate. The temperature of the biotransformation system is controlled at 37° C.; the pH value of the biotransformation system is controlled at 6.5, and the mechanical stirring speed is controlled at 200 r / min.

[0059] The conversion solution containing L-hydroxyproline is purified, comprising the following steps:

[006...

Embodiment 3

[0066] L- Biotransformation method of hydroxyproline , the reaction was carried out in a 50L fermenter, and the reaction system was controlled to 30L, including 2400g of L-proline and 800g of α-ketoglutarate as substrates, and 800g of proline-4-hydroxylase engineering The whole cell of the bacteria is used as a catalyst, and the biotransformation system is established by adding surfactants and additives to carry out the transformation reaction, and the transformation liquid containing L-hydroxyproline is obtained. Wherein, the surfactant consists of 12.8g of sorbitan ester, 32g of Tween 80, and 3.2g of Triton X-100. The additive is a mixture of 3.2g ferrous sulfate, 4g ferrous chloride and 0.8g ferrous nitrate. The temperature of the biotransformation system is controlled at 40° C.; the pH value of the biotransformation system is controlled at 8, and the mechanical stirring speed is controlled at 400 r / min.

[0067] The conversion solution containing L-hydroxyproline is pu...

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Abstract

The present invention discloses a bioconversion method of L-hydroxyproline. According to the bioconversion method, a surfactant and an additive are added to establish a bioconversion system by using L-proline and alpha-ketoglutaric acid as substrates and using the whole cell of engineering bacteria containing proline-4-hydroxylase as a catalyst, and a conversion reaction is performed to obtain the conversion liquid containing L-hydroxyproline. According to the present invention, the bioconversion method has advantages of strong stereoselectivity, mild reaction conditions, easy operation, low cost, less pollution and less catalyst consumption, and can complete the reactions difficultly performed in chemical reactions, wherein the space time yield is more than 62.36 g / d / L, the yield is more than 90%, the crystal purity is more than 98%, and the optical purity is more than 99%.

Description

technical field [0001] The present invention relates to a preparation method of amino acid derivatives, in particular to a L- Biotransformation method of hydroxyproline . Background technique [0002] Hydroxyproline (Hyp) is the main component of collagen and does not belong to the 20 common amino acids. In recent years, the research and development of L-hydroxyproline has attracted extensive attention from the fields of medicine, biochemistry, food and beauty. It can be used as a cosmetic additive, with anti-oxidation and anti-radiation effects; it also has weight-loss effects, and is expected to become an ideal weight-loss drug; It can also speed up wound healing and treat various skin diseases. As an important component of amino acid injection, L-hydroxyproline has a certain curative effect on hypoalbuminemia caused by acute and chronic liver diseases; it participates in the emulsification of fat and the formation of erythrocyte heme and globulin, and has the function...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/24C12R1/19
CPCC12P13/24
Inventor 易菊阳刘立明陈建华潘崇德
Owner 绍兴百茵生物技术有限公司
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