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Method for producing R-alpha-hydroxybutyrate by using 1, 2-butanediol as substrate

A technology of hydroxybutyric acid and butanediol, which is applied in the field of producing R-alpha-hydroxybutyric acid, can solve the problems that the method for producing R-alpha-hydroxybutyric acid has not been reported, and achieves easy separation and purification, simple reaction system, Inexpensive effect

Active Publication Date: 2013-05-15
上海肆芃科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the current retrieval results show that the method for producing R-α-hydroxybutyric acid using Gluconobacter oxydans has not been reported

Method used

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  • Method for producing R-alpha-hydroxybutyrate by using 1, 2-butanediol as substrate
  • Method for producing R-alpha-hydroxybutyrate by using 1, 2-butanediol as substrate
  • Method for producing R-alpha-hydroxybutyrate by using 1, 2-butanediol as substrate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] (1) Preparation of biocatalyst

[0043] Gluconobacter oxidans DSM 2003 (purchased from the German Collection of Microorganisms) was selected, cultured by conventional methods, the bacteria were separated and collected, and the bacteria were washed with pH 7.4 potassium phosphate buffer for 3 times, and the bacteria were suspended in deionized water to make the bacteria concentration When reaching 200 g wet cells / liter, the obtained complete cell suspension is the biocatalyst, stored at 4°C for later use;

[0044] Among them, the above-mentioned Gluconobacter oxydans is cultivated using the following medium formula: D-sorbitol 80 g / L; yeast powder 24 g / L; ammonium sulfate 5 g / L; potassium dihydrogen phosphate 2 g / L; magnesium sulfate 5 g / L Liter; Calcium carbonate 10 g / liter.

[0045] (2) Conversion

[0046] The biocatalyst prepared in step (1), ie, intact cells, is mixed with the substrate 1,2-butanediol aqueous solution, so that the concentration of the biocatalyst in the mix...

Embodiment 2

[0052] (1) Preparation of biocatalyst

[0053] Select Gluconobacter oxydans DSM 2003, culture by conventional methods, separate and collect the bacteria, wash the bacteria 3 times with pH 7.4 potassium phosphate buffer, and suspend the bacteria in deionized water to make the bacteria concentration reach 200 g wet cells / liter. The obtained complete cell suspension is the biocatalyst, stored at 4°C for later use;

[0054] Among them, the above-mentioned Gluconobacter oxydans is cultivated using the following medium formula: D-sorbitol 80 g / L; yeast powder 24 g / L; ammonium sulfate 5 g / L; potassium dihydrogen phosphate 2 g / L; magnesium sulfate 5 g / L Liter; Calcium carbonate 10 g / liter.

[0055] (2) Conversion

[0056] Mix the biocatalyst prepared in step (1), i.e. intact cells, with the substrate 1,2-butanediol aqueous solution, so that the concentration of the biocatalyst in the mixture is 100 g wet cells / liter, 1,2-butanediol The concentration is 50 g / L, and disodium edetate is added ...

Embodiment 3

[0062] (1) Preparation of biocatalyst

[0063] Select Gluconobacter oxydans DSM 2003, culture by conventional methods, separate and collect the bacteria, wash the bacteria 3 times with pH 7.4 potassium phosphate buffer, and suspend the bacteria in deionized water to make the bacteria concentration reach 200 g wet cells / liter. The obtained complete cell suspension is the biocatalyst, stored at 4°C for later use;

[0064] Among them, the above-mentioned Gluconobacter oxydans is cultivated using the following medium formula: D-sorbitol 80 g / L; yeast powder 24 g / L; ammonium sulfate 5 g / L; potassium dihydrogen phosphate 2 g / L; magnesium sulfate 5 g / L Liter; Calcium carbonate 10 g / liter.

[0065] (2) Conversion

[0066] The biocatalyst prepared in step (1), ie, intact cells, is mixed with the substrate 1,2-butanediol aqueous solution, so that the concentration of the biocatalyst in the mixture is 120 g wet cells / liter, 1,2-butanediol The concentration is 60 g / L, and disodium edetate is ad...

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Abstract

The invention discloses a method for producing 1,2-butanediol by using R-alpha-hydroxybutyrate as a substrate. The method comprises steps of: culturing Gluconobacter oxydans DSM 2003 by a conventional method to prepare a biological catalyst; mixing the biological catalyst with a substrate 1,2-butylene glycol aqueous solution; adding ethylene diamine tetraacetic acid and oscillating for 1-30 h under condition of 20-50 DEG C and pH of 4.0-10.0 to obtain a transformation liquid; and preparing a solution containing R-alpha-hydroxybutyrate through the transformation liquid. The method provided by the invention has the following characteristics: (1) the method employs biological catalysis, has simple reaction system, mild reaction conditions, short steps and simple operation; and the biological catalyst can be easily removed, so as to facilitate subsequent separation and purification; (2) the method has a short reaction period, and the product R-alpha-hydroxybutyrate can accumulate to a high concentration; (3) the substrate 1,2-butanediol has low price, and is easy to acquire; and (4) product enantiomer has high excessive rate reaching higher than 99%, so as to lay foundation for the efficient production of R-alpha-hydroxybutyrate.

Description

Technical field [0001] The present invention relates to a method for producing R-α-hydroxybutyric acid, in particular to a method that uses intact cells of Gluconobacter oxydans DSM 2003 as a biocatalyst to catalyze the production of R-α-hydroxybutyrate by 1,2-butanediol The sour method. Background technique [0002] Alpha-hydroxybutyric acid is an important industrial intermediate, which can be used to synthesize isoleucine and certain drugs. α-Hydroxybutyric acid is divided into R and S configurations. High purity chiral R-α-hydroxybutyric acid can be used to synthesize biodegradable polymer polyα-hydroxybutyric acid [P(2HB)] 【1】 . In addition, R-α-hydroxybutyric acid can be used to prepare azinothricin family anticancer antibiotics 【2，3】 . [0003] After literature search, there are usually two kinds of starting substrates for the production of high-purity R-α-hydroxybutyric acid. One is α-ketobutyric acid, which generates R-α-hydroxybutyrate through stereoselective enzymati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/42C12R1/01
Inventor 许平高超张文马翠卿
Owner 上海肆芃科技有限公司
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