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Method for preparing D-amino acid and alpha keto acid

A technology of alpha keto acid and amino acid, applied in the field of biology

Inactive Publication Date: 2018-12-28
浙江正硕生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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There are other methods that have certain limitations

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  • Method for preparing D-amino acid and alpha keto acid
  • Method for preparing D-amino acid and alpha keto acid
  • Method for preparing D-amino acid and alpha keto acid

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Embodiment Construction

[0037] A preparation method of D-amino acid and alpha keto acid, comprising the steps of:

[0038] 1. Construction of recombinant bacteria:

[0039] Total gene synthesis from Proteus mirabilis ( Proteus mirabilis) L-amino acid deaminase (L-aminoacid deaminase, LAAD) gene (genbank accession number EU669819.1) LAAD, and connected into the pET24a vector ( NdeI / Hind ), construct the plasmid LAAD, transform it into E.Coli BL21 (DE3), and construct the recombinant strain PMLAAD.

[0040] Total gene synthesis is derived from lactic acid bacteria ( Lactobacillus buchneri strain JCM 1115) amino acid racemase (amino acid racemase, AAR) gene (genbank accession number KC413940.1) AAR, and connected into the pET24a vector ( NdeI / Hind ), construct the plasmid AAR, transform it into E.Coli BL21 (DE3), and construct the recombinant strain AAR.

[0041] 2. Fermentation

[0042] Recombinant bacteria PMLAAD and AAR were fermented with TB.

[0043] 3) Medium

[0044] (1) LB:

...

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Abstract

The invention belongs to the field of biotechnology, and relates to a method for preparing D-amino acid and alpha keto acid. The method includes the first step of recombinant construction: conductingwhole gene synthesis to obtain an L-amino acid deaminase gene LAAD and connecting an expression vector to be transformed into an expression strain to construct a recombinant PMLAAD; conducting whole gene synthesis to obtain an amino acid racemase gene AAR and connecting the expression vector to be transformed into the expression strain to construct a recombinant AAR; the second step of fermentation; the third step of enzyme catalysis; the fourth step of isolation and purification. The alpha-keto acid calcium and the D-amino acid are produced by enzyme catalysis, and both products have economicvalue and high product quality, so that the production cost is greatly reduced.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a preparation method of D-amino acid and alpha keto acid. Background technique [0002] There are 20 kinds of amino acids in nature, all of which are L-type amino acids, which are the basic structural units of proteins. Therefore, it is generally believed that D-amino acids rarely exist in nature. In recent years, with the deepening of scientific research, it has been found that although D-amino acids are not the basic structural nitrogen source of proteins, they are found in many plants, microorganisms and higher plants. Presence of D-amino acids. [0003] At present, the application fields of D-amino acid are mainly: 1. Medicine. For the synthesis of peptide drugs, such as peptide antibiotics (Apoxicillin ASPOXICILLIN), gastrointestinal drugs (OXTREOTIDE), diuretics, diarrhea drugs (SANDOSTATINACETATE), enzyme inhibitors, corticotropin analogs, analgesic drugs, weight loss Drugs, ...

Claims

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Application Information

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IPC IPC(8): C12P13/08C12P13/06C12P13/22C12P7/40C12N1/21C12R1/19
CPCC12N9/0022C12N9/90C12P7/40C12P13/06C12P13/08C12P13/222C12Y104/03002C12Y501/0101
Inventor 吴黎诚郭小雷章权
Owner 浙江正硕生物科技有限公司
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