Significance of the 20-kDa Subunit of Heterodimeric 2-Deoxy-scyllo-inosose Synthase for the Biosynthesis of Butirosin Antibiotics in Bacillus circulans(Microbiology & Fermentation Technology)

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概要

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• A gene (btrC2) encoding the 20-kDa subunit of 2-deoxy-scyllo-inosose (DOI) synthase, a key enzyme in the biosynthesis of 2-deoxystreptamine, was identified from the butirosin-producer Bacillus circulans by reverse genetics. The deduced amino acid sequence of BtrC2 closely resembled that of YaaE of B. subtilis, but the function of the latter has not been known to date. Instead, BtrC2 appeared to show sequence similarity to a certain extent with HisH of B. subtilis, an amidotransferase subunit of imidazole glycerol phosphate synthase. Disruption of btrC2 reduced the growth rate compared with the wild type, and simultaneously antibiotic producing activity was lost. Addition of NH_4Cl to the medium complemented only the growth rate of the disruptant, and both the growth rate and antibiotic production were restored by addition of yeast extract. In addition, a heterologous co-expression system of btrC2 with btrC was constructed in Escherichia coli. The simultaneously over-expressed BtrC2 and BtrC constituted a heterodimer, the biochemical features of which resembled those of DOI synthase from B. circulans more than those of the recombinant homodimeric BtrC. Despite the similarity of BtrC2 to HisH the heterodimer showed neither aminotransfer nor amidotransfer activity for 2-deoxy-scyllo-inosose as a substrate. All the observations suggest that BtrC2 is involved not only in the secondary metabolism, but also in the primary metabolism in B. circulans. The function of BtrC2 in the butirosin biosynthesis appears to be indirect, and may be involved in stabilization of DOI synthase and in regulation of its enzyme activity.

• 社団法人日本農芸化学会の論文
• 2002-07-23

著者

• Tamegai Hideyuki

  Department of Chemistry, College of Humanities and Sciences, Nihon University

• KAKINUMA Katsumi

  Department of Chemistry, Tokyo Institute of Technology

• KUDO Fumitaka

  Department of Chemistry, Tokyo Institute of Technology

• Kakinuma Katsumi

  Tokyo Institute of Technology, Department of Chemistry

• NANGO Eriko

  Department of Chemistry, Tokyo Institute of Technology

• KOIKE-TAKESHITA Ayumi

  Department of Chemistry, Tokyo Institute of Technology

• Kakinuma K

  Tokyo Inst. Technol. Tokyo

• Kakinuma Katsumi

  Department Of Chemistry And Materials Science Tokyo Institute Of Technology

• Kakinuma Katsumi

  Department Of Chemistry Tokyo Inst. Of Tech.

• Koike-takeshita Ayumi

  Department Of Chemistry Tokyo Institute Of Technology

• Koike‐takeshita A

  Bio Research Laboratory Toyota Motor Corporation

• Kudo Fumitaka

  Department Of Chemistry Tokyo Institute Of Technology

• Kudo Fumitaka

  Department Of Orthopedic Surgery Kyorin University School Of Medicine

• Kakinuma K

  Tokyo Inst. Technology Tokyo Jpn

• Tamegai Hideyuki

  Department Of Chemistry College Of Humanities And Sciences Nihon University

• Nango Eriko

  Department Of Chemistry Graduate School Of Science And Engineering Tokyo Institute Of Technology

• Kakinuma Katsumi

  Department of Chemistry & Materials Science, and Department of Chemistry, Tokyo Institute of Technology

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