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Accelerated Growth of Corynebacterium glutamicum by Up-Regulating Stress- Responsive Genes Based on Transcriptome Analysis of a Fast-Doubling Evolved Strain

Journal of Microbiology and Biotechnology 2020년 30권 9호 p.1420 ~ 1429
박지훈, 이수린, 이민주, 박경훈, 이승기, 김지현, 김필,
소속 상세정보
박지훈 ( Park Ji-Hoon ) - Catholic University Department of Biotechnology
이수린 ( Lee Su-Rin ) - Catholic University Department of Biotechnology
이민주 ( Lee Min-Ju ) - Catholic University Department of Biotechnology
박경훈 ( Park Kyung-Hoon ) - Catholic University Department of Biotechnology
이승기 ( Lee Seung-Ki ) - Catholic University Department of Biotechnology
김지현 ( Kim Ji-Hyun ) - Yonsei University Department of Systems Biology
김필 ( Kim Pil ) - Catholic University Department of Biotechnology

Abstract


Corynebacterium glutamicum, an important industrial strain, has a relatively slower reproduction rate. To acquire a growth-boosted C. glutamicum, a descendant strain was isolated from a continuous culture after 600 generations. The isolated descendant C. glutamicum, JH41 strain, was able to double 58% faster (td=1.15 h) than the parental type strain (PT, td=1.82 h). To understand the factors boosting reproduction, the transcriptomes of JH41 and PT strains were compared. The mRNAs involved in respiration and TCA cycle were upregulated. The intracellular ATP of the JH41 strain was 50% greater than the PT strain. The upregulation of NCgl1610 operon (a putative dyp-type heme peroxidase, a putative copper chaperone, and a putative copper importer) that presumed to role in the assembly and redox control of cytochrome c oxidase was found in the JH41 transcriptome. Plasmid-driven expression of the operon enabled the PT strain to double 19% faster (td=1.82 h) than its control (td=2.17 h) with 14% greater activity of cytochrome c oxidase and 27% greater intracellular ATP under the oxidative stress conditions. Upregulations of genes those might enhance translation fitness were also found in the JH41 transcriptome. Plasmid-driven expressions of NCgl0171 (encoding a cold-shock protein) and NCgl2435 (encoding a putative peptidyl-tRNA hydrolase) enabled the PT to double 22% and 32% faster than its control, respectively (empty vector: td=1.93 h, CspA: td=1.58 h, and Pth: td=1.44 h). Based on the results, the factors boosting growth rate in C. gluctamicum were further discussed in the viewpoints of cellular energy state, oxidative stress management, and translation.

키워드

Corynebacterium glutamicum; adaptive laboratory evolution; rapid reproduction; cellular energy; oxidative stress management; translation process

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