Bbd29_14255 gene mutant and application thereof in preparing l-glutamic acid

Abstract

The application discloses a BBD29_14255 gene mutant and application thereof in preparation of L-glutamic acid. The BBD29_14255 gene mutant disclosed by the application is a DNA molecule shown in SEQ ID No. 3, 5, 7 in the sequence listing, encodes a protein shown in SEQ ID No. 4, 6, 8, and a wild type BBD29_14255 gene is a DNA molecule shown in SEQ ID No. 1, encodes a protein shown in SEQ ID No. 2. Experiments prove that the BBD29_14255 gene and the mutant thereof can improve the yield of L-glutamic acid, can be used for producing L-glutamic acid, and have a good application prospect.

Claims

1. Protein, as shown in A1 or A2): A1) A mutant protein obtained by mutating the leucine residue at position 172 of SEQ ID No. 2 to a phenylalanine residue, a valine residue, or an isoleucine residue; A2) is a fusion protein obtained by attaching a tag to the N-terminus and / or C-terminus of A1).

2. The biological material related to the protein of claim 1 is any one of B1) to B4) below: B1) A nucleic acid molecule encoding the protein of claim 1; B2) An expression cassette containing the nucleic acid molecule described in B1); B3) A recombinant vector containing the nucleic acid molecule described in B1), or a recombinant vector containing the expression cassette described in B2); B4) Recombinant microorganisms containing the nucleic acid molecules described in B1), or recombinant microorganisms containing the expression cassette described in B2), or recombinant microorganisms containing the recombinant vector described in B3).

3. The biomaterial of claim 2, wherein: B1) The nucleic acid molecule described is as follows (b11), (b12), or (b13): b11) The DNA molecule shown in SEQ ID No. 3 of the sequence listing; b12) The DNA molecule shown in SEQ ID No. 5 of the sequence listing; b13) The DNA molecule shown in SEQ ID No. 7 of the sequence listing.

4. The biomaterial according to claim 2 or 3, characterized in that: B2) The expression cassette is the DNA molecule shown in SEQ ID No. 10, SEQ ID No. 11, or SEQ ID No. 12; B4) The recombinant microorganism is a recombinant microorganism obtained by replacing the BBD29_14255 gene in a microorganism containing the BBD29_14255 gene shown in SEQ ID No. 1 with SEQ ID No. 3, SEQ ID No. 5 or SEQ ID No. 7, or a recombinant microorganism obtained by introducing the nucleic acid molecule described in B1) into a microorganism and causing it to be expressed.

5. A method of producing L-glutamic acid, comprising: The protein of claim 1 is expressed in the recipient biological cells, or the content or activity of the protein shown in SEQ ID No. 4, SEQ ID No. 6, SEQ ID No. 8 or SEQ ID No. 2 is increased in the recipient biological cells to obtain recombinant biological cells; the recombinant biological cells are cultured to obtain L-glutamic acid.

6. The method according to claim 5, characterized in that: The biological cells are bacteria, algae, fungi, plant cells, or animal cells that can synthesize L-glutamic acid.

7. The method according to claim 6, characterized in that: The fungus is yeast; the bacteria is Corynebacterium glutamicum.

8. The method according to any one of claims 5-7, characterized in that: The method is achieved by introducing the gene encoding the protein shown in SEQ ID No. 4, SEQ ID No. 6, SEQ ID No. 8, or SEQ ID No. 2 into the recipient biological cell and allowing it to be expressed; Alternatively, the recipient biological cell contains the DNA molecule shown in SEQ ID No. 1, and the method is implemented by replacing the DNA molecule shown in SEQ ID No. 1 in the recipient biological cell with the DNA molecule shown in SEQ ID No. 3, SEQ ID No. 5, or SEQ ID No.

7.

9. The method according to any one of claims 5-7, characterized in that: The recombinant biological cells are cultured using a culture medium that enables the recombinant biological cells to grow. And / or, the recombinant biological cells are cultured under conditions that enable the recombinant biological cells to grow.

10. A product for preparing L-glutamic acid, comprising the protein of claim 1 or the biomaterial of any one of claims 2-4.

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