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Method for preparing S-(+)-3-methyl hydroxybutyrate through microbial transformation

A technology for the transformation of methyl hydroxybutyrate and microorganisms, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of cumbersome catalyst preparation and high cost, and achieve easy large-scale industrial production, simple operation, The effect of mild reaction conditions

Inactive Publication Date: 2011-05-25
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] A large number of literature reports use various types of nickel catalysts to catalyze the asymmetric hydrogenation reaction of methyl acetoacetate to prepare methyl S-(+)-3-hydroxybutyrate. In this process, the preparation of the catalyst is cumbersome and the cost is high.

Method used

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  • Method for preparing S-(+)-3-methyl hydroxybutyrate through microbial transformation
  • Method for preparing S-(+)-3-methyl hydroxybutyrate through microbial transformation
  • Method for preparing S-(+)-3-methyl hydroxybutyrate through microbial transformation

Examples

Experimental program
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Effect test

Embodiment 1

[0027] Saccharomyces cerevisiae CGMCC No. 2266 strain was inoculated into the slant medium, and cultured at 30° C. for 4 to 6 days to prepare the bacterial cell slant. Use an inoculation needle to take an inoculation loop from the slant of the bacterium 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 a seed solution. Inoculate 10 mL of seed solution (the inoculum size is 10% of the volume of the liquid medium) into a 250 mL Erlenmeyer flask containing 100 mL of liquid medium, cultivate it for 24 hours at 30° C. and 180 r / min to obtain a fermented liquid, and centrifuge the fermented liquid to obtain Enzyme cells.

[0028] The determination of the dry weight of the bacteria is to take a small part from the wet cells after centrifuging the fermentation broth and dry it at 120°C for 48 hours to a constant weight, measure the weight of the dry cells, and calculate the ratio of dry cells in the ...

Embodiment 2

[0036] Saccharomyces cerevisiae CGMCC No.2266 strain was inoculated into the slant medium, and cultured at 30° C. for 4 to 6 days to obtain the slant surface of the bacteria. Use an inoculation needle to take an inoculation loop from the slant of the bacterium 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 a seed solution. The seed solution was inoculated in a 250mL Erlenmeyer flask containing 100mL of liquid medium with a 10% inoculation amount, cultivated at 30°C and 180r / min for 24 hours to obtain a fermentation broth, and the fermentation broth was centrifuged to obtain enzyme-containing bacterial cells. The dry weight of enzyme-containing bacterium cells contained in 1 liter of fermented liquid is 50 grams.

[0037] In five parts of Erlenmeyer flasks containing 100mL pH7.0 phosphate buffer solution, add the wet cells obtained after centrifugation of 200 milliliters of fermented liquid...

Embodiment 3

[0042] Saccharomyces cerevisiae CGMCC No.2266 strain was inoculated into the slant medium, and cultured at 30° C. for 4 to 6 days to obtain the slant surface of the bacteria. Use an inoculation needle to take an inoculation loop from the slant of the bacterium 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 a seed solution. The seed solution was inoculated in a 250mL Erlenmeyer flask containing 100mL of liquid medium with a 10% inoculation amount, cultivated at 30°C and 180r / min for 24 hours to obtain a fermentation broth, and the fermentation broth was centrifuged to obtain enzyme-containing bacterial cells. The dry weight of enzyme-containing bacterium cells contained in 1 liter of fermented liquid is 50 grams.

[0043] In five parts of Erlenmeyer flasks containing 100mL pH7.0 phosphate buffer solution, add the wet cells obtained after centrifugation of 200 milliliters of the above-mentio...

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Abstract

The invention provides a method for preparing S-(+)-3-methyl hydroxybutyrate through microbial transformation, which comprises the following steps that: in phosphate buffer solution with the pH value being 5.0 to 8.0, methyl acetoacetate is used as substrates, enzyme-containing strains obtained through fermenting saccharomyces cerevisiae CGMCC No. 2266 are used as microbial catalysts, conversion region is carried out for 8 to 40h at 25 to 45 DEG C, after the reaction is completed, and conversion liquid is prepared into the S-(+)-3-methyl hydroxybutyrate through separation and purification. The method provided by the invention has the main beneficial effects that 1. produced strains are safe and innoxious and are easy to be cultured in a large scale; 2. a large number of biological catalysts can be obtained, and the cost is low; 3. the reaction conditions are mild, and the environmental-friendly effect is realized; 4. the operation is simple and convenient, and the addition of coenzymes with a high price is not needed in the reaction process; and 5. the large-scale industrialized production is easy to realize, and the mol conversion rate is high.

Description

(1) Technical field [0001] The invention relates to a method for preparing methyl S-(+)-3-hydroxybutyrate by microbial transformation, in particular to a method using Saccharomyces cerevisiae CGMCC No.2266 as a biocatalyst and using methyl acetoacetate as a base Preparation of S-(+)-3-hydroxybutyrate methyl ester method. (2) Background technology [0002] S-(+)-3-hydroxybutyrate methyl ester ((S)-(+)-methyl 3-hydroxybutyrate), CAS accession number: 53562-86-0, molecular formula C 5 h 10 o 3 , molecular weight 118.13. Can be used for the synthesis of optically active poly-β-hydroxybutyrate P[(R)-HB], P[(R)-HB] is a kind of biopolyester in bacterial cells, which has good biodegradability, and its decomposition The products can all be used biologically without any pollution to the environment. Its melting temperature is 175-180°C, and it is a thermoplastic that can be completely decomposed. It can be used as a chiral intermediate for drug synthesis. [0003] S-(+)-methyl ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/62C12R1/865
Inventor 欧志敏陈庆美隋志红
Owner ZHEJIANG UNIV OF TECH
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