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Method for producing D-a amino acid series by using De's bacterium of Bakehuo in onions

A technology of Burkholderia bacterium and JS-02, which is applied in the direction of bacteria and fermentation, and can solve the problems of low production yield, D-phenylalanine production cost cannot meet the application needs, complex process and other problems

Inactive Publication Date: 2003-07-23
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above preparation methods or processes are complicated and the preparation yield is low, or the transition metal catalyst used is expensive, resulting in that the preparation cost of D-phenylalanine cannot meet the application needs

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Take 20.0g of glucose, 15.0g of corn steep liquor, 3.0g of NaCl, MgSO 4 ·7H 2 O0.5g, KH 2 PO 4 1.0g, CoCl 2 0.1g, N-carbamoyl phenylalanine 1.5g, add water 1000ml, adjust PH7.0-7.2, dissolve and put into 250ml triangular flasks, each bottle contains 50ml, then add 8 layers of gauze, wrap with kraft paper , 0.1MPa high-pressure steam sterilization for 25 minutes for later use.

[0037] Take out the strains stored in the refrigerator, put them on a fresh slant, cultivate them for 24 hours, insert them into the liquid culture medium prepared above, shake them at 180 rpm at 32°C for 24 hours, then separate them at 5000 rpm Centrifuge for 25 minutes, collect the bacteria, and make a bacteria suspension (every 100ml of bacteria suspension contains about 40 grams of wet bacteria).

[0038] Take 3.0g of DL-5-benzylhydantoin and put it into the prepared 500ml Erlenmeyer flask, and add 100ml of deionized water respectively to adjust the pH to 9.0, and then add 10ml of bacter...

Embodiment 2

[0040] Culture medium composition and disinfection conditions are the same as Example 1. After culturing for 14 hours, the strains were inserted into the prepared liquid culture medium, shaken at 30°C at 180 rpm for 30 hours, centrifuged at 5000 rpm for 25 minutes, collected bacteria, and made into a bacterial suspension (per 100ml bacterial suspension contains about 40 grams of wet bacterial cells).

[0041] Take 4.0g of DL-5-indolmethylhydantoin and put it into the prepared 500ml Erlenmeyer flask, and add 100ml of deionized water respectively to adjust the pH to 9.0, and then add 10ml of bacterial suspension to each Erlenmeyer flask. Replace the oxygen in the space of the Erlenmeyer flask with nitrogen, add a rubber stopper, and shake the reaction on a shaker. The reaction times were 42 hours, respectively. After the reaction, centrifuge at 4000 rpm for 30 minutes to remove bacteria and impurities, and concentrate under vacuum at 55-60°C. After being washed with alcohol, ...

Embodiment 3

[0043] Culture medium composition and disinfection conditions are the same as Example 1. After culturing for 18 hours, the strains were inserted into the prepared liquid medium, shaken at 180 rpm at 32°C for 30 hours, centrifuged at 5,000 rpm for 30 minutes, and the bacteria were collected to make a bacterial suspension (per 100ml bacterial suspension contains about 40 grams of wet bacterial cells).

[0044] Take 3.0g of DL-5-methylthioethylhydantoin and put it into the prepared 500ml Erlenmeyer flask, add 100ml of deionized water respectively, adjust the pH to 9.0, and then add 10ml of bacterial suspension into each Erlenmeyer flask. Replace the oxygen in the space of the Erlenmeyer flask with nitrogen, add a rubber stopper, and shake the reaction on a shaker. The reaction times were 36 hours, respectively. After the reaction, centrifuge at 4000 rpm for 25 minutes to remove bacteria and impurities, and concentrate under vacuum at 55-60°C. After washing with alcohol, the cr...

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PUM

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Abstract

A process for preparing the optically active D-alpha-amino acid series (D-Phe, D-Trap, D-Val, D-methionie, etc) from 5-substituted hydrantoin features use of Burkholderia cepecia JS-02 (CCTCC No.M202047) as splitting catalyst, which is obtained from soil. Its advantages are high output rate and high concentration of substrate.

Description

technical field [0001] D-a-amino acid is a class of optically active compounds that are widely used in the fields of medicine, food and pesticides. For example, D-phenylalanine, D-tryptophan, D-tyrosine and D-methionine are important components of synthetic polypeptide drugs; D-alanine is a key component of synthetic sweetener alitame; D-valine is an important component in the preparation of optically active insecticides, etc. Background technique [0002] The current synthetic method of this kind of substance usually adopts the chemical resolution method of the racemate. Taking D-phenylalanine as an example, the current preparation method is as follows: first, DL-phenylalanine is prepared by chemical synthesis, and L-phenylalanine and N-acetyl- D-phenylalanine, N-acetyl-D-phenylalanine obtained after separation is hydrolyzed at low temperature under the action of dilute acid to obtain D-phenylalanine; another method of asymmetric synthesis is used to prepare D-phenylalani...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P13/04
Inventor 韦萍周华于荣华李环万红贵胡永红欧阳平凯
Owner NANJING UNIV OF TECH
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