Characterization and Functional Modification of StaC and RebC, Which Are Involved in the Pyrrole Oxidation of Indolocarbazole Biosynthesis
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概要
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The diversity of indolocarbazole natural products results from the differences in oxidation states of the pyrroline ring moiety. In the biosynthetic pathways for staurosporine and rebeccamycin, two homologous enzymes having 64% identity, StaC and RebC, are responsible for the selective production of K252c, which has one oxo group at the pyrroline ring, and arcyriaflavin A, which has two. Although StaC has a FAD-binding motif, most StaC molecules do not contain FAD, and the protein cannot be reconstituted with FAD in vitro. In this study, we mutated Ala-118 in StaC by replacing a glutamine that is conserved in FAD monooxygenases, resulting in increased FAD content as well as catalytic activity. In addition, mutations around the substrate-binding sites of StaC and RebC can change the product selectivity. Specifically, StaC-N244R-V246T and RebC-F216V-R239N mutants produced substantial amounts of arcyriaflavin A and K252c, respectively.
- 2011-11-23
著者
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Otani H
Faculty Of Agriculture Tottori University
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IGARASHI Yasuhiro
Biotechnology Research Center, Toyama Prefectural University
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Nagano Shingo
Biometal Science Laboratory Riken Spring-8 Center
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Igarashi Yasuhiro
Department Of Mechanical Engineering The University Of Tokyo
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Onaka H
Department Of Engineering Toyama Prefectural University
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Onaka Hiroyasu
Dep. Of Biotechnology Fac. Of Engineering And Biotechnology Res. Center Toyama Prefectural Univ.
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Onaka Hiroyasu
Department Of Biotechnology Division Of Agriculture And Life Sciences The University Of Tokyo
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Shiro Yoshitsugu
Biometal Sci. Lab. Riken Spring-8 Center
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Asamizu Shumpei
Department Of Biotechnology Faculty Of Engineering Toyama Prefectural University
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Igarashi Yasuhiro
Department Of Biotechnology Faculty Of Engineering Toyama Prefectural University
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Shiro Yoshitsugu
Biometal Science Laboratory Riken Spring-8 Center
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Igarashi Y
Biotechnology Research Center Toyama Prefectural University
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