Saccharomyces cerevisiae and application thereof in preparing (R)-(+)-beta-hydroxy-benzenepropanoic acid ethylester by microbial transformation

A technology of ethyl carbonyl phenylpropionate and Saccharomyces cerevisiae, which is applied to Saccharomyces cerevisiae and the application field in the preparation of (R)-(+)-β-hydroxyphenylpropionate ethyl ester by microbial transformation, and can solve the problem of product yield and Problems such as unsatisfactory enantioselectivity, to achieve the effects of easy large-scale industrial production, high biotransformation rate, and simple production and operation

Inactive Publication Date: 2010-05-19
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] According to the reports and studies on the synthesis of (R)-(+)-beta-hydroxyphenylpropionate by microbial methods at present, the yield and enantioselectivity of the product are not ideal, and it is necessary to find a catalyst with higher catalytic efficiency. strains, and select appropriate reaction conditions to maximize product yield and enantiomeric excess

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Slant culture: Inoculate CGMCC No.3361 bacteria into the slant medium and culture at 30°C for 4-6 days. The slant medium is prepared as follows: wort juice 10g / L, yeast powder 3g / L, peptone 5g / L, glucose 10g / L, agar 20g / L, natural pH, solvent is water; sterilized at 121°C 20min, after sterilizing, cool to make a slope.

[0028] Seed culture and fermentation: Both the seed and the fermentation medium use liquid medium, which is prepared according to the following composition: glucose 30g / L, yeast powder 3g / L, ammonium sulfate 5g / L, anhydrous MgSO 4 0.25g / L, K 2 HPO 4 ·3H 2 O 1g / L, KH 2 PO 4 1g / L, sterilized at 121°C for 20min. The pH value of the liquid medium was adjusted to 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0 with 1mol / L NaOH or 1mol / L HCl respectively.

[0029] Use an inoculation needle to take a ring of bacteria from the slant culture medium and inoculate it into a 250ml Erlenmeyer flask containing 100ml of liquid medium, and culture it at 30°C and 1...

Embodiment 2

[0035] Slant culture: Inoculate CGMCC No.3361 bacteria into the slant medium and culture at 30°C for 4-6 days. The slant medium is prepared as follows: wort juice 10g / L, yeast powder 3g / L, peptone 5g / L, glucose 10g / L, agar 20g / L, natural pH, solvent is water; sterilized at 121°C 20min, after sterilizing, cool to make a slope.

[0036] Seed culture and fermentation: Both the seed and the fermentation medium use liquid medium, which is prepared according to the following composition: glucose 30g / L, yeast powder 3g / L, ammonium sulfate 5g / L, anhydrous MgSO 4 0.25g / L, K 2 HPO 4 ·3H 2 O 1g / L, KH 2 PO 4 1g / L, sterilized at 121°C for 20min. Adjust the pH of the liquid medium to 7.0.

[0037] Use an inoculation needle to take a ring of bacteria from the slant culture medium and inoculate it into a 250ml Erlenmeyer flask containing 100ml of liquid medium, and culture it at 30°C and 180r / min for 24h to obtain seeds. The seed liquid was inoculated into a 250 ml Erlenmeyer flask ...

Embodiment 3

[0043] Slant culture: Inoculate CGMCC No.3361 bacteria into the slant medium and culture at 30°C for 4-6 days. The slant medium is prepared as follows: wort juice 10g / L, yeast powder 3g / L, peptone 5g / L, glucose 10g / L, agar 20g / L, natural pH, solvent is water; sterilized at 121°C 20min, after sterilizing, cool to make a slope.

[0044] Seed culture and fermentation: Both the seed and the fermentation medium use liquid medium, which is prepared according to the following composition: glucose 30g / L, yeast powder 3g / L, ammonium sulfate 5g / L, anhydrous MgSO 4 0.25g / L, K 2 HPO 4 ·3H 2 O 1g / L, KH 2 PO 4 1g / L, sterilized at 121°C for 20min. Adjust the pH of the liquid medium to 7.0.

[0045] Use an inoculation needle to take a ring of bacteria from the slant culture medium and inoculate it into a 250ml Erlenmeyer flask containing 100ml of liquid medium, and culture it at 30°C and 180r / min for 24h to obtain seeds. The seed liquid was inoculated into a 250 ml Erlenmeyer flask ...

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Abstract

The invention provides a new strain-Saccharomyces cerevisiae CGMCC NO.3361 and application thereof in preparing (R)-(+)-beta-hydroxy-benzenepropanoic acid ethylester by microbial transformation. The Saccharomyces cerevisiae was collected in China General Microbiological Culture Collection Center (CGMCC), the address of which is Institute of Microbiology, Chinese Academy of Sciences, Datun Road, Chaoyang District, Beijing 100101. The collection date is October 23, 2009 and the collection number is CGMCC NO.3361. The beneficial effects of preparing (R)-(+)-beta-hydroxy-benzenepropanoic acid ethylester by the microbial transformation method in the invention are mainly as follows: (1) the biocatalyst is a microbial thallus, thus is lower in cost than the chemical catalysts; (2) the environment is friendly, the reaction conditions are mild and the transformation can be carried out smoothly under normal temperature and normal pressure; (3) the strain used in the production is safe and nontoxic; (4) the method is not influenced by seasons and is easy to realize large-scale industrial production; (5) the production operation is simple and convenient and the biotransformation rate is higher; and (6) coenzyme is unnecessary to be added in the whole reaction process.

Description

(1) Technical field [0001] The invention relates to a new bacterial strain—Saccharomyces cerevisiae CGMCC No.3361, and its application in the preparation of (R)-(+)-beta-hydroxyphenylpropionate ethyl by microbial transformation. (2) Background technology [0002] (R)-(+)-β-hydroxyphenylpropanoic acid ethyl ester (Benzenepropanoic acid, β-hydroxy, ethylester, (βR)-), molecular formula C 11 h 14 o 3 , molecular weight 194.23, CAS number: 72656-47-4. Ethyl (R)-(+)-β-hydroxyphenylpropionate is an important intermediate in the synthesis of (R)-atomoxetine. Atomoxetine is an inhibitor that inhibits the absorption of norepinephrine. Atomoxetine has a high affinity for norepinephrine absorption sites. It can maintain the concentration of norepinephrine in the nervous system to a certain level, which can improve people's endurance and antidepressant. The efficacy of (R)-atomoxetine is twice that of racemic atomoxetine and 9 times that of (S)-atomoxetine. [0003] (R)-(+)-β-Hyd...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/16C12P7/62C12R1/865
Inventor 欧志敏陈晓彦应国清
Owner ZHEJIANG UNIV OF TECH
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