• Title/Summary/Keyword: Mycothiol

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Identification of Genes for Mycothiol Biosynthesis in Streptomyces coelicolor A3(2)

  • Park Joo-Hong;Cha Chang-Jun;Roe Jung-Hye
    • Journal of Microbiology
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    • v.44 no.1
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    • pp.121-125
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    • 2006
  • Mycothiol is a low molecular weight thiol compound produced by a number of actinomycetes, and has been suggested to serve both anti-oxidative and detoxifying roles. To investigate the metabolism and the role of mycothiol in Streptomyces coelicolor, the biosynthetic genes (mshA, B, C, and D) were predicted based on sequence homology with the mycobacterial genes and confirmed experimentally. Disruption of the mshA, C, and D genes by PCR targeting mutagenesis resulted in no synthesis of mycothiol, whereas the mshB mutation reduced its level to about $10\%$ of the wild type. The results indicate that the mshA, C, and D genes encode non-redundant biosynthetic enzymes, whereas the enzymatic activity of MshB (acetylase) is shared by at least one other gene product, most likely the mca gene product (amidase).

Cloning and Expression of Inositol Monophosphatase Gene from Streptomyces coelicolor A[3]2 (Streptomyces coelicolor A[3]2에서 Mycothiol 생합성에 관여하는 Inositol Monophosphatase 유전자의 클로닝 및 발현)

  • Kim Jin Kwon;Choi Hack Sun;Kim Seong-Jun;Kim Si Wouk
    • KSBB Journal
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    • v.19 no.6 s.89
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    • pp.462-466
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    • 2004
  • Mycothiol (MSH), a low molecular antioxidant thiol compound, was purified and analyzed from Streptomyns coelicolor A[3]2 by the monobromobimane fluorescence detection method modified by this lab. Through HPLC chromatpgram, MSH fraction was obtained following the elution time of standard MSH (donated by Dr. Robert C. Fahey). That MSH showed the highest concentration among the thiol compounds contained in the cell indicated that MSH was the key thiol compound having antioxidant activity. To understand the role of gene of inositol monophosphatase (I-1-Pase) involved in the MSH biosynthesis, it was isolated from S. coelicolor A(3)2 and cloned and overexpressed in the Escherichia coli. The expressed I-1-Pase was purified through Ni-NTA column. The soluble protein consisted of 281 amino acids, and the molecular weight was 32 kDa. I-1-Pase of S. coelicolor A(3)2 had the sequence homology with those of human and E. coli by 24 and $25\%$, respectively, and had two conserved domains (mofif A and motif B) which were typical of I-1-Pase.

Comparative Genomics Reveals the Core and Accessory Genomes of Streptomyces Species

  • Kim, Ji-Nu;Kim, Yeonbum;Jeong, Yujin;Roe, Jung-Hye;Kim, Byung-Gee;Cho, Byung-Kwan
    • Journal of Microbiology and Biotechnology
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    • v.25 no.10
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    • pp.1599-1605
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    • 2015
  • The development of rapid and efficient genome sequencing methods has enabled us to study the evolutionary background of bacterial genetic information. Here, we present comparative genomic analysis of 17 Streptomyces species, for which the genome has been completely sequenced, using the pan-genome approach. The analysis revealed that 34,592 ortholog clusters constituted the pan-genome of these Streptomyces species, including 2,018 in the core genome, 11,743 in the dispensable genome, and 20,831 in the unique genome. The core genome was converged to a smaller number of genes than reported previously, with 3,096 gene families. Functional enrichment analysis showed that genes involved in transcription were most abundant in the Streptomyces pan-genome. Finally, we investigated core genes for the sigma factors, mycothiol biosynthesis pathway, and secondary metabolism pathways; our data showed that many genes involved in stress response and morphological differentiation were commonly expressed in Streptomyces species. Elucidation of the core genome offers a basis for understanding the functional evolution of Streptomyces species and provides insights into target selection for the construction of industrial strains.