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Mutant gene and coding protein of glutamate dehydrogenase of Bacillus natto

A technology of glutamate dehydrogenase and Bacillus natto, which is used in genetic engineering, plant genetic improvement, oxidoreductase and other directions

Inactive Publication Date: 2011-07-27
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

Examples

Experimental program
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Embodiment

[0028] Step 2, determine the active site of Bacillus natto GDH and the site-directed mutagenesis scheme, use the SAMM molecular simulation analysis software to perform energy minimization analysis on the protein model, and obtain the possible interaction with the substrate glutamic acid (Glu ) have amino acid residues connected by hydrogen bonds, and perform mutation simulation on several amino acid residues with a higher probability of occurrence; the details are as follows:

[0029] ClustalX: Using ClustalX, using 1GTM, 1EUZ, 1PZO, 1HWY, 1V9L, and 2RIR as the comparison objects, multiple sequence alignments of Bacillus subtilis GDH were obtained, and four highly consistent amino acid sequences were obtained, respectively: 80 - Residue 82, Residue 112-120, Residue 195-200, Residue 351-358, the four sequences are very close to the spatial distance of the substrate glutamic acid The region of the enzyme activity center is likely to be contained in it;

[0030] SAMM: Use the S...

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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 fin...

Claims

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Application Information

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Patent Type & Authority Patents(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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