Method for catalytically preparing tanshinol through planococcus rifietoensis

A technology of danshensu and cocci, which is applied in the field of bioengineering, can solve the problems of expensive catalysts and retention

Inactive Publication Date: 2016-12-07
汕头市佳禾生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The catalyst for chiral synthesis of Danshensu (patent CN201210420488.4) is extremely expensive, and currently only stays at the laboratory level

Method used

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  • Method for catalytically preparing tanshinol through planococcus rifietoensis

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Analytical Determination of Transformation Products.

[0023] Prepare medium as follows: glucose 20g / L, yeast yeast extract 5g / L, peptone 10g / L, sodium chloride 5g / L. The liquid volume in the 500ml Erlenmeyer flask is 100ml, a total of 50 bottles, sterilized at 120°C for 20 minutes. Take Planococcus rifietoensisATCC BAA-790 glycerol tube seed solution 1mL to inoculate, the fermentation temperature is 30°C, and the shaker speed is 200rpm. After culturing for 24 hours, the fermentation broth was centrifuged (rotating speed 10000rpm, time 10min), the supernatant was removed to obtain bacterial cells, the bacterial cells obtained by centrifugation were washed once with sterile water and then centrifuged (rotating speed 10000rpm, time 10min) to obtain wet bacterial cells. Get 20g of wet thallus and put it into 1000ml solution with cinnamic acid concentration of 2g / L, and mix well. Shake (100 rpm) at 35°C for 24 hours, then heat in a water bath at 100°C for 3 minutes to kil...

Embodiment 2

[0027] The medium composition is: fructose 50g / L, ammonium sulfate 5g / L. The volume of liquid in a 250ml Erlenmeyer flask is 50ml, sterilized at 120°C for 20 minutes. Take Planococcus rifietoensis ATCC BAA-790 glycerol tube seed solution 0.5mL to inoculate, the fermentation temperature is 20°C, and the shaker speed is 200rpm. After culturing for 72 minutes, the fermentation broth was centrifuged (10,000 rpm, 10 min), the supernatant was removed to obtain bacterial cells, and the centrifuged bacterial cells were washed once with sterile water, and then centrifuged (10,000 rpm, 10 min) to obtain wet bacterial cells. Get the wet thallus and put it into the 2ml solution whose L-dopa concentration is 2g / L, so that the thallus concentration reaches 1g / L, and mix well. After shaking (100 rpm) on a shaking table at 20° C. for 72 hours, the converted solution was filtered through a microporous membrane, and the concentration of danshensu in the converted solution was analyzed by liqui...

Embodiment 3

[0029] The medium composition is: sucrose 25g / L, ammonium sulfate 1g / L, diammonium hydrogen phosphate 1g / L, potassium dihydrogen phosphate 0.5g / L, magnesium sulfate 0.2g / L, ferrous sulfate 0.1g / L. The volume of liquid in a 250ml Erlenmeyer flask is 50ml, sterilized at 120°C for 20 minutes. Take Planococcus rifietoensis ATCC BAA-790 glycerol tube seed solution 0.5mL to inoculate, the fermentation temperature is 35°C, and the shaker speed is 200rpm. After culturing for 48 hours, the fermentation broth was centrifuged (10,000 rpm, 10 min), the supernatant was removed to obtain bacterial cells, and the centrifuged bacterial cells were washed once with sterile water, and then centrifuged (10,000 rpm, 10 min) to obtain wet bacterial cells. Take 0.05g of wet bacteria and put it into 2ml solution with cinnamaldehyde concentration of 2g / L, and mix well. After shaking (100 rpm) on a shaking table at 20° C. for 24 hours, the converted solution was filtered through a microporous membrane...

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Abstract

The invention discloses a method for producing R-(+)-3-(3,4-dihydroxyphenyl)-2-hydracrylic acid (D-tanshinol) through planococcus rifietoensis, and belongs to the technical field of bioengineering. By means of planococcus rifietoensis, D-tanshinol is catalytically prepared from substrates (cinnamic acid, cinnamyl aldehyde, cinnamyl alcohol, L-DOPA, L-tyrosine and L-phenylalanine). Specifically, the catalysis process A of adding the substrates to fermentation liquid in the liquid fermentation process or the catalysis process B of obtaining cells through liquid fermentation culture and putting the cells in a reaction system to catalyze the substrates is adopted. The method has good industrial application prospects.

Description

technical field [0001] The preparation of danshensu by using the catalyzed substrate of Lepitoplankton belongs to the technical field of bioengineering. The invention relates to a method for preparing danshensu with a catalyzed substrate of Lepitoplankton sp., and the invention is of great significance to the industrial production of danshensu. Background technique [0002] Danshensu, scientific name R-(+)-3-(3,4-dihydroxyphenyl)-2-hydroxypropionic acid, D-(+)-β-(3,4-dihydroxyphenyl)lactic acid, English Name: Danshensu, (R)-(+)-3-(3,4-Dihydroxyphenyl)-lactic acid, (R)-(+)-3-(3,4-Dihydroxyphenyl)-2-hydroxypropanoic acid, is A dextrophenolic acid compound. [0003] Danshensu is an important active ingredient in the water extract of Danshen, and its structure was obtained and identified from the water extract of Danshen in 1980 in China (Study on the water-soluble active ingredients of Danshen, Ⅱ.D(+)β(3,4- The structure of dihydroxyphenyl) lactic acid, Shanghai First Medica...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/42C12R1/01
CPCC12P7/42
Inventor 蔡宇杰金大勇管政兵丁彦蕊杨波廖祥儒
Owner 汕头市佳禾生物科技有限公司
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