阅读说明：本技术 发酵生产l-赖氨酸的方法 (Method for producing L-lysine by fermentation ) 是由 F·施耐德 G·蒂尔巴赫 K·福斯 T·贝克尔 于 2019-07-24 设计创作，主要内容包括：本发明提供了使用具有分泌L-赖氨酸的能力的谷氨酸棒杆菌(Corynebacterium glutamicum)物种的细菌发酵生产L-赖氨酸的新方法,在所述细菌的染色体中含有编码具有水解酶活性的多肽的多核苷酸的变体。(The present invention provides a novel method for the fermentative production of L-lysine using a bacterium of the species Corynebacterium glutamicum (Corynebacterium glutamicum) having the ability to secrete L-lysine, which contains in its chromosome a variant of a polynucleotide encoding a polypeptide having hydrolase activity.)

1. A method for producing L-lysine by fermentation, comprising the following steps:

a) providing a bacterium of the species Corynebacterium glutamicum (Corynebacterium glutamicum) having the ability to secrete L-lysine, comprising in its chromosome a polynucleotide encoding a polypeptide comprising the amino acid sequence of SEQ ID NO. 4,

b) culturing the bacterium under suitable conditions in a suitable medium, and

c) accumulating the L-lysine in the medium to form an L-lysine containing fermentation broth.

2. The method of claim 1, wherein in the provided bacterium, the polynucleotide encoding the amino acid sequence comprises the nucleotide sequence of positions 923 to 2812 of SEQ ID NO 3.

3. The method of claim 1, wherein in the provided bacterium, the polynucleotide encoding the amino acid sequence comprises the nucleotide sequence of positions 923 to 2815 of SEQ ID NO. 3.

4. The method of claim 1, wherein in the provided bacteria, the polynucleotide encoding the amino acid sequence comprises the nucleotide sequence of positions 585 to 2815 of SEQ ID NO 3.

5. The method of claim 1, wherein in the provided bacteria, the polynucleotide encoding the amino acid sequence comprises the nucleotide sequence of positions 565 to 2815 of SEQ ID No. 3.

Example 1

Sequence of NCgl1480 gene of Corynebacterium glutamicum strain DM1933

The "", in the name NCgl1480, indicates that the coding sequence of NCgl1480 comprises a sequence encoding 26 additional amino acids at the 5' end (see SEQ ID NO:1 and SEQ ID NO:2) in comparison with the information given under log _ tag NCgl1480, according to the present invention.

Strain DM1933 is the L-lysine producer described by Blombach et al (Applied and environmental Microbiology75(2),419-427, 2009). It is deposited according to the budapest bar at DSMZ under accession number DSM 25442.

The nucleotide sequence of the chromosome of strain DM1933 was determined by Illumina whole genome sequencing technology (Illumina inc., San Diego, CA, US). See, e.g., Benjak et al (2015) white-Genome sequencing for comprehensive Genome and De Novo Genome Assembly. in: Parish T., Roberts D. (eds) Mycobacterium protocols in Molecular Biology, Vol 1285 HumanaPress, NY, US) and Bennet, S. (Pharmacogenomics 5(4),433-438, 2004).

The nucleotide sequence of the NCgl1480 coding sequence of the strain DM1933 (including the nucleotide sequences upstream and downstream thereof) was found to be identical to the nucleotide sequence of ATCC13032 shown in SEQ ID NO: 1.

DM1933 contains in its chromosome a variant of the aspartokinase gene encoding a feedback resistant aspartokinase polypeptide. The feedback resistant aspartokinase polypeptide has an amino acid sequence of SEQ ID NO 6 of the sequence Listing, wherein the amino acid threonine (Thr) at amino acid sequence position 311 is replaced by isoleucine (Ile). In US7,338,790, the abbreviation "lysC T311I" is used to indicate the exchange. Blombach et al uses the abbreviation "lysC (T311I)".