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Mutant gene and coding protein of glutamate dehydrogenase of Bacillus natto

A technology of glutamate dehydrogenase and Bacillus natto, which can be used in genetic engineering, plant genetic improvement, oxidoreductase and other directions, and can solve problems that have not yet been reported.

Inactive Publication Date: 2010-09-08
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] finds through literature retrieval to prior art, does not see the report relevant with the theme of the present invention as yet

Method used

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  • Mutant gene and coding protein of glutamate dehydrogenase of Bacillus natto
  • Mutant gene and coding protein of glutamate dehydrogenase of Bacillus natto
  • Mutant gene and coding protein of glutamate dehydrogenase of Bacillus natto

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Embodiment

[0024] Step 1, homologous modeling of Bacillus natto GDH; first search for a protein sequence highly similar to the GDH protein (SEQ ID NO: 2) sequence in the PDB, try to model on the Swiss model, and use the Prosa2003 residue After the energy map analysis, the 1GTM protein from Pyrococcus furiosus and the 1EUZ protein from Mesophilic coccus were finally selected as templates for homology modeling, and the model was modeled, and the obtained protein model was optimized by using SAMM molecular simulation analysis software; the specific process is as follows :

[0025] Using NCBI's BLASTn to compare and analyze the obtained Bacillus natto GDH coding gene sequence (SEQ ID NO: 1), use the ORF Finder module to search for possible open reading frames in the sequence, and find one containing 422 Proteins of amino acid residues were compared and analyzed in BLASTp;

[0026] Six highly similar protein sequences (about 50% similarity) to the protein sequence were obtained in PDB, namel...

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Abstract

The invention relates to a mutant gene and coding protein of glutamate dehydrogenase of Bacillus natto, belonging to the field of biotechnology. The base sequence of the mutant gene of the coding gene of glutamate dehydrogenase of Bacillus natto is shown as follows: SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13 or SEQ ID NO: 15; the protein amino acid sequence of the mutant gene of the coding gene of glutamate dehydrogenase of Bacillus natto is shown as follows: SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14 or SEQ ID NO: 16. The invention also relates to a recombinant vector containing the mutant gene. When the mutation of the invention is introduced to wild strains by homologous recombination, gene engineering bacteria with low ammonia yield and unchanged growth speed is obtained, thus producing natto without ammonia odor.

Description

technical field [0001] The invention relates to a gene and encoded protein in the field of biotechnology, in particular to a mutant gene and encoded protein of the Bacillus natto glutamic acid dehydrogenase gene. Background technique [0002] Bacillus Subtilis Natto is a fermentation bacterium of Japanese natto, which can produce a variety of unique health-care ingredients after fermentation, such as nattokinase, polyglutamic acid, vitamin K 2 etc., nattokinase has been confirmed to have a good thrombolytic function, and no toxic side effects. In addition, natto also has physiological prevention and regulation functions such as preventing osteoporosis, anti-oxidation, antibacterial, and regulating the stomach. Except that the prevention of osteoporosis is related to the isoflavones contained in soybean itself, other functions mainly depend on the new substances produced after fermentation. The Japanese regard July 10th as the Natto Festival. However, natto is hard to find...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/53C12N9/04C12N15/63
Inventor 张建华陈丽丽王家乐
Owner SHANGHAI JIAO TONG UNIV
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