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Purification and characterization of an arginine regulatory protein, ArgR, in Corynebacterium glutamicum

  • Original Paper
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Journal of Industrial Microbiology & Biotechnology

Abstract

Corynebacterium glutamicum, a Gram-positive bacterium, has been widely used for industrial amino acid production. We previously showed that, in C. glutamicum, argCJBDFRGH arginine biosynthesis genes are clustered but independently transcribed from argC and argG promoters, leading to the generation of two transcripts corresponding to argCJBDFR and argGH. In this report, we show the effect of the C. glutamicum ArgR repressor on argC and argG promoters by overexpressing or disrupting the argR gene. Gel filtration assay results indicate that native ArgR is a hexamer of equal subunits with molecular mass of 110 kDa. Protein sequence analysis revealed the presence of an “SR” (Ser57-Arg58) motif for the DNA binding site at the N-terminal region and the “GTIAGDDTV” motif for arginine binding and its oligomerization at the C-terminal region. An argC or argG promoter–lacZ fusion reporter assay and argR mutational analysis showed that transcription of the argCJBDFR arginine biosynthesis genes is regulated from the argC promoter by ArgR in cooperation with l-arginine in C. glutamicum. This finding was supported by the gel mobility–shift assay showing direct binding of hexameric ArgR to the argC promoter in the presence of l-arginine. Unexpectedly, argGH transcription was not responsive to the level of ArgR repressor and/or arginine. In a further study, a C. glutamicum argR mutant was constructed by disrupting the chromosomal argR gene to manufacture an improved arginine-producing strain. Arginine productivity was increased in the C. glutamicum argR mutant strain under conditions of both limited and excessive arginine.

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Acknowledgments

This work was supported by Sookmyung Women’s University (2010).

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Authors and Affiliations

  1. Division of Biological Science and Research Center for Women’s Diseases, Sookmyung Women’s University, Seoul, 140-742, South Korea

    Sei-Hyun Yim, Samil Jung, Shun-kyoung Lee, Choong-Il Cheon, Eunsook Song & Myeong-Sok Lee

  2. Samsung Advanced Institute of Technology, Suwon, 440-600, South Korea

    Soo-Suk Lee

  3. Songdo Technopark, 7-50 Songdo-dong, Yeonsu-gu, Inchon, 406-840, South Korea

    Jin Shin

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  1. Sei-Hyun Yim
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Correspondence to Myeong-Sok Lee.

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S.-H. Yim and S. Jung contributed equally to this paper.

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Yim, SH., Jung, S., Lee, Sk. et al. Purification and characterization of an arginine regulatory protein, ArgR, in Corynebacterium glutamicum . J Ind Microbiol Biotechnol 38, 1911–1920 (2011). https://doi.org/10.1007/s10295-011-0977-9

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