Method for preparing L-2-aminobutyric acid through biocatalysis

A technology of biocatalysis and aminobutyric acid, applied in the field of medicine and chemical industry, can solve the problems of high price

Active Publication Date: 2015-09-30
ABA CHEM CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the route of synthesizing levetiracetam through a series of reactions such as methylation, esterification, and amidation, using L-2-aminobutyric acid as the key starting material, although the reaction conditions are mild, side reactions are few, and the product quality is high , and the total yield is high, but the price of the raw material L-2-aminobutyric acid is expensive, therefore, the synthesis technology of L-2-aminobutyric acid has always been a focus of pharmaceutical engineering research in recent years

Method used

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  • Method for preparing L-2-aminobutyric acid through biocatalysis
  • Method for preparing L-2-aminobutyric acid through biocatalysis
  • Method for preparing L-2-aminobutyric acid through biocatalysis

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Experimental program
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Effect test

Embodiment 1

[0045] A kind of method that biocatalysis prepares L-2-aminobutyric acid comprises the following steps:

[0046] The first step, heterologous expression of formate dehydrogenase FDH and leucine dehydrogenase LDH:

[0047] (a) Heterologous expression and activity measurement of FDH:

[0048] (1) Obtain FDH gene fragment

[0049] Primers were designed according to the gene sequence of formate dehydrogenase (FDH) (GenBank accession numbers: XM_001525495 and AAB18593) and its front and back sequences, and the better upstream primer of formate dehydrogenase (FDH) obtained after screening was 5'-AAA CATATG AAAATCGTTCTCGTTTTGTACTCC-3' (the underline is the NdeI restriction site, SEQ ID NO.: 1); the downstream primer is 5'-AAA CTCGAG TGCGACCTTTTTTGTCATTAC-3' (the underline is the XhoI restriction site, SEQ ID NO.: 2). FDH gene fragments can be efficiently obtained by amplifying with the above-mentioned upstream and downstream primers.

[0050] (2) PCR reaction system

[0051] Th...

Embodiment 2

[0076] The steps are all the same as in Example 1, except that the reaction temperature in the second step is 25°C. The yield of the product obtained was 53.5%.

Embodiment 3

[0078] The steps are all the same as in Example 1, except that the reaction temperature in the second step is 28°C. The yield of the product obtained was 65.8%.

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Abstract

The invention provides a method for preparing L-2-aminobutyric acid through biocatalysis. The method comprises the following steps: establishing genetically engineered bacteria for simultaneously expressing FDH (Formate Dehydrogenase) and LDH (Leucine Dehydrogenase) to obtain an FDH/LDH co-expression crude enzyme solution containing the FDH and the LDH through fermentation; preparing a reaction system in a buffer solution, and performing biocatalytic reaction to prepare L-2-aminobutyric acid, wherein the reaction system comprises threonine, ammonium formate, a TD enzyme solution, the FDH/LDH co-expression crude enzyme solution, NAD<+> and pyridoxal phosphate. Through the adoption of the method, the preparation of L-2-aminobutyric acid can be simple, efficient and low in cost.

Description

technical field [0001] The invention relates to the field of medicine and chemical industry, in particular to a method for preparing L-2-aminobutyric acid by biocatalysis. Background technique [0002] L-2-aminobutyric acid is a key intermediate for the production of the new antiepileptic drug levetiracetam, and it is also a key chiral precursor for the synthesis of the antibacterial and anti-tuberculosis drug ethambutol. It has become an important chiral intermediate of various chiral drugs. Therefore, the synthesis technology of L-2-aminobutyric acid has always been a hot spot in pharmaceutical engineering research in recent years. [0003] Reduced coenzyme I (nicotinamide adenine dinucleotide or nicotinic acid diphosphate, NADH) is an important coenzyme in redox reactions. As an electron donor, one molecule of NADH can provide two electrons for the reduction of groups such as ketone groups. With the development of biocatalytic technology, NADH is increasingly used in c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/04C12N9/04C12N9/06C12N15/70
CPCC12P13/04
Inventor 李航徐军阙利民江岳恒蔡彤
Owner ABA CHEM CORP
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