Production improvement of antifungal, antitrypanosomal nucleoside sinefungin by rpoB mutation and optimization of resting cell system of Streptomyces incarnatus NRRL8089(MICROBIAL PHYSIOLOGY AND BIOTECHNOLOGY)
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概要
- 論文の詳細を見る
Sinefungin, a nucleoside antibiotic with potent antifungal, antiviral, and anti-trypanosome activities, has been a target for production enhancement in the past decades through medium optimization and strain improvement. For the purpose of introducing a more rational approach, we induced rpoB mutation in the producer strain, Streptomyces incarnatus NRRL 8089, by optimized UV-irradiation, and a resulting rifampicin-resistant strain rif-400 increased the sinefungin production by 7-fold. The growth and melanin production were obviously accelerated in the rifampicin-resistant high-producer mutant, while the morphological differentiation such as aerial mycelia and spiked-spore formation was retained. Molecular cloning and DNA sequencing identified a single mutation A1340G in the rpoB gene, which encodes the β-subunit of RNA polymerase, and the resulting amino acid substitution Asp447Gly corresponded to one of mutations that reportedly allowed the transcriptional up-regulation of actinorhodin production in S. coelicolor A3(2). Sinefungin production was further enhanced by resting cell system using the rpoB mutant strain in the presence of 10mM L-Arg. D-Arg or L-ornithine did not enhance the sinefungin production, and >50mM urea strongly suppressed the nucleoside antibiotic production, supporting the proposed biosynthetic mechanism by which urea is liberated from the guanidino-group-bearing intermediate that is produced by enzymatic condensation of L-Arg and ATP.
著者
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FUKUDA Koji
Department of Cardiovascular Medicine, Tohoku University Graduate School of Medicine
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Tamura Takashi
Department of Bioresources Chemistry, Graduate School of Natural Science and Technology, Okayama Uni
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Ito Hideyuki
Division of Pharmaceutical Sciences, Graduate School of Medicine, Dentistry, and Pharmaceutical Scie
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Yamamoto Sayaka
Department of Bioresources Chemistry, Graduate School of Natural Science and Technology, Okayama Uni
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Ochi Kozo
National Food Research Institute
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Inagaki Kenji
Department of Bioresources Chemistry, Graduate School of Natural Science and Technology, Okayama Uni
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Ochi Kozo
National Food Res. Inst.
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Furutani Y
Okayama Univ. Okayama Jpn
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Inagaki Kenji
Dep. Of Bioresources Chemistry Graduate School Of Natural Sci. And Technol. Okayama Univ. 1-1-1 Tsus
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Inagaki Kenji
Department Of Bioresources Chemistry Graduate School Of Natural Science And Technology Okayama Unive
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Inagaki Kenji
Department Of Biofunctional Chemistry Graduate School Of Natural Science And Technology Okayama Univ
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Tamura T
Department Of Biofunctional Chemistry Graduate School Of Natural Science And Technology Okayama Univ
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Yamamoto Sayaka
Department Of Bioresources Chemistry Graduate School Of Natural Science And Technology Okayama Unive
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Yamamoto Sayaka
Department Of Applied Life Science Faculty Of Engineering Sojo University
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Tamura Takashi
Department Of Applied Chemistry Faculty Of Science Science University Of Tokyo
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Ito Hideyuki
Division Of Pharmaceutical Sciences Graduate School Of Medicine Dentistry And Pharmaceutical Science
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Inagaki Kenji
Department Of Bioresources Chemistry Graduate School Of Natural Science And Technology Okayama Unive
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Fukuda Koji
Department Of Bioresources Chemistry Graduate School Of Natural Science And Technology Okayama Unive
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Inagaki K
Department Of Bioresources Chemistry Graduate School Of Natural Science And Technology Okayama Unive
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INAGAKI KENJI
Research Institute for Wakan-Yaku, Toyama Medical and Pharmaceutical University
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