Mining and preliminary verification of genes involved in the high heterologous protein production of Corynebacterium glutamicum based on whole-genome resequencing
MENG Lihong1,2,3, LIU Xiuxia1,2,3*, YANG Yankun1,2,3, BAI Zhonghu 1,2,3
1(Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China) 2(National Engineering Laboratory of Cereal Fermentation Technology, Jiangnan University, Wuxi 214112, China) 3(School of Biotechnology, Jiangnan University, Wuxi 214122, China)
Abstract: Food-safe strain Corynebacterium glutamicum can be used as an excellent chassis cell for industrial production of high-value products. It is a potential host for the expression of recombinant proteins due to its good secretion system. In order to optimize the chassis cells and improve their heterologous protein expression ability, genes in C. glutamicum which can highly express heterologous protein were mined and verified. On the basis of whole-genome resequencing of C. glutamicum, bioinformatics was used to mine single-nucleotide polymorphisms(SNP) genes and their conserved domains. The recombinant strains with overexpression or knockout of SNP genes were constructed to evaluate their(SNP) growth and the expression ability of heterologous proteins EGFP and rtPTH. The resequencing data of high heterologous protein production strains were mined. There were 33 SNPs, mapped to five genes. Among them, SNP recombinant strains over-2370 and ko-973-974 had the highest fluorescence intensity, and the production of rtPTH by fermentation was nearly two-folds higher than that of wild type. This study has successfully mined genes involved in the high yield of heterologous protein, GL002370 and GL000974. These findings will help to obtain optimized chassis cells and provide guidance for in-depth research on target genes that can increase protein production.
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2021, Vol. 47 
