A Strain of Bacillus subtilis Producing High-Yield L-lysine: Construction

doi:10.11924/j.issn.1000-6850.casb20191000699

Abstract

Abstract:

To breed a strain producing L-lysine with non-resistance, high yield, good genetic stability, low production cost, pepper anther and Bacillus subtilis were used as test materials, the shuttle expression vector pHT43 and gene knockout vector pK18mobsacB was performed as a vector to transform lysine-rich protein gene (CFLR) into Bacillus subtilis. After two rounds of self-recombination and screening of culture medium, the final strain CMCC (B) 63501/pK18mobsacB-ΔALA::CFLR, with high yield of L-lysine was obtained without antibiotic resistance. After 20 generations of serial passage, the strain still expressed a significant higher number of CFLR copies than the wild type. The subsequent data of High-Performance Liquid Chromatography (HPLC) revealed that the content of L-lysine in recombinant strain was 64.89% higher than that of the wild strain. The above results indicated that the recombinant strain we constructed were featured with non-resistance, high yield, good genetic stability and low purification cost. Therefore, the strain exhibits outstanding potential for future applications.

Key words: CFLR gene, Bacillus subtilis, gene cloning, homologous exchange, HPLC

CLC Number:

Xia Shuli, Han Jing, Wang Lixue, Yang Chunlei, Liu Changyue, Wang Hailiang, Wu Kuang, Shi Huiling. A Strain of Bacillus subtilis Producing High-Yield L-lysine: Construction[J]. Chinese Agricultural Science Bulletin, 2020, 36(30): 98-105.

Figures/Tables 9

References 21

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