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Method for producing L-2-aminobutyric acid

A production method, the technology of aminobutyric acid, applied in the biological field, can solve the problems of high cost, unstable properties of butyronic acid, and restrictions on industrial application, and achieve the effects of low process cost, high conversion rate and product concentration

Active Publication Date: 2014-02-05
湖州颐盛生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This process adopts butanuonic acid as raw material, due to the high price of butanuonic acid, the price of 5g of butanuonic acid on the market is more than 100 yuan, resulting in high cost price of the process, and the property of butanuonic acid is unstable, which limits the application of the process. Industrial application

Method used

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  • Method for producing L-2-aminobutyric acid
  • Method for producing L-2-aminobutyric acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 The reaction system of threonine deaminase derived from Escherichia coli, leucine dehydrogenase derived from Bacillus cereus and glucose dehydrogenase derived from Bacillus subtilis catalyzes the production of L-2-aminobutyric acid

[0026] Prepare liquid medium TB (pH7.0-7.5), containing 12g / L peptone, 24g / L imported yeast extract, 5g / L glycerol, 2.13g / L KH 2 PO 4 , 16.43g / L K 2 HPO 4 ·3H 2 O. Medium TB was divided into 1L Erlenmeyer shake flasks with a volume of 200mL, and then heated and sterilized in an autoclave at 121°C for 20min.

[0027] Leucine dehydrogenase expression strains derived from Bacillus cereus (refer to Kula MR, Stoyan T, Recktenwald A. Cloning, sequencing and overexpression of the leucine dehydrogenasegene from Bacillus cereus. Journal of Biotechnology. 1997, 54: 77 -80 described method construction), Bacillus subtilis derived glucose dehydrogenase expression bacterial species (with reference to Lampel KA, et al. 43) and Escherichia...

Embodiment 2

[0033] Example 2 The reaction system of threonine deaminase derived from Escherichia coli, leucine dehydrogenase derived from Bacillus cereus and formate dehydrogenase derived from Candida boidinii catalyzes the production of L-2-aminobutyric acid

[0034] The strain fermentation, the preparation of the crude enzyme solution and the liquid phase detection method are the same as in Example 1, wherein, the formate dehydrogenase expression strain derived from Candida boidinii refers to Sakai Y, et al. Regulation of the formatedehydrogenase gene, FDH1 , in the methylotrophic yeast Candida boidinii and growth characteristics of an FDH1-disrupted strain on methanol, methylamine, and choline. JBacteriol.1997, 179 (14): 4480-5 described method construction.

[0035] The enzyme activity detection method of threonine deaminase derived from Escherichia coli and leucine dehydrogenase derived from Bacillus cereus is the same as that in Example 1.

[0036] The detection method of formate de...

Embodiment 3

[0038] Example 3 Valine dehydrogenase derived from Streptomyces albicans, threonine deaminase derived from Escherichia coli and glucose dehydrogenase derived from Bacillus subtilis catalyzed by the reaction system to produce L-2-aminobutyric acid

[0039] Strain fermentation, preparation of crude enzyme solution and liquid phase detection method are the same as in Example 1, wherein, the expression strain of valine dehydrogenase derived from Streptomyces albicans refers to Hyuu CG, Kim SS et al.Valine dehydrogenase from Streptomyces albus gene cloning heterologous expression and identification of active site by site-directed mutagenesis. FEMS Microbiology Letters. 2000, 182: 29-34 described method construction.

[0040] The enzyme activity detection method of threonine deaminase derived from Escherichia coli and glucose dehydrogenase derived from Bacillus subtilis is the same as that in Example 1.

[0041] The detection method of valine dehydrogenase enzyme activity derived fr...

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Abstract

The invention discloses a method for producing L-2-aminobutyric acid, and the method comprises the following step: catalyzing L-threonine utilized as a raw material through an enzyme catalysis system consisting of threonine deaminase, L-amino acid dehydrogenase and coenzyme regeneration systems, thus producing the L-2-aminobutyric acid. The method for producing theL-2-aminobutyric acid has the advantages that the raw material is low in price, the property is stable, and the production cost of the L-2-aminobutyric acid can be greatly lowered, the conversion rate and product concentricity are high, no influence caused by byproducts exists, and the method is suitable for industrialization application.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a production method of L-2-aminobutyric acid. Background technique [0002] L-2-aminobutyric acid, the English name is L(+)-2-Aminobutyric acid, the molecular formula is C4H9NO2, and the CAS No. is 1492-24-6. It is a non-natural chiral α-amino acid and is a new type of anti-epileptic Medicine - the main raw material for the production of levetiracetam. Levetiracetam is a new type of anti-epileptic drug developed by UCB (UCB) in Belgium. It was approved by the FDA in April 2000 and listed in the United States and the European Union. It is mainly used for the treatment of localized and secondary dysentery. generalized epilepsy. It is currently on the market in more than 66 countries and regions around the world, and has more than 1 million treatment records worldwide. It is currently the most widely used new antiepileptic drug in the treatment of epilepsy in the United St...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P13/04
Inventor 杨晟范文超陶荣盛朱傅赟沈正权陆沈高陈涛潘震华
Owner 湖州颐盛生物科技有限公司
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