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ID 48634
本文ファイル
著者
Takahashi, Hitoshi 大学院医歯薬学総合研究科
Kumagai, Takanori 大学院医歯薬学総合研究科 広大研究者総覧
Kitani, Kyoko 大学院医歯薬学総合研究科
Mori, Miwako 大学院医歯薬学総合研究科
Matoda, Yasuyuki 大学院医歯薬学総合研究科
Sugiyama, Masanori 大学院医歯薬学総合研究科 広大研究者総覧
抄録(英)
In the present study, we cloned a gene, designated bpsA, which encodes a single module type non-ribosomal peptide synthetase (NRPS) from a d-cycloserine (DCS)-producing Streptomyces lavendulae ATCC11924. A putative oxidation domain is significantly integrated into the adenylation domain of the NRPS, and the condensation domain is absent from the module. When S. lividans was transformed with a plasmid carrying bpsA, the transformed cells produced a blue pigment, suggesting that bpsA is responsible for the blue pigment synthesis. However, to produce the blue pigment in Escherichia coli, the existence of the 4′-phosphopantetheinyl transferase (PPTase) gene from Streptomyces was necessary, in addition to bpsA. The chemical structure of the pigment was determined as 5,5′-diamino-4,4′-dihydroxy-3,3′-diazadiphenoquinone-(2,2′), called indigoidine. The bpsA gene product, designated BPSA, was overproduced in an E. coli host-vector system and purified to homogeneity, demonstrating that the recombinant enzyme prefers l-Gln as a substrate. The in vitro experiment using l-Gln also showed that the blue pigment was formed by the purified BPSA only when the enzyme was phosphopantetheinylated by adding a Streptomyces PPTase purified from E. coli cells. Each site-directed mutagenesis experiment of Lys598, Tyr601, Ser603, and Tyr608, which are seen in the oxidation domain of BPSA, suggests that these residues are essential for the binding of FMN to the protein and the synthesis of the blue pigment.
掲載誌名
Journal of Biological Chemistry
282巻
12号
開始ページ
9073
終了ページ
9081
出版年月日
2007-03-23
出版者
The American Society for Biochemistry and Molecular Biology, Inc.
ISSN
0021-9258
1083-351X
出版者DOI
言語
英語
NII資源タイプ
学術雑誌論文
広大資料タイプ
学術雑誌論文
DCMIタイプ
text
フォーマット
application/pdf
著者版フラグ
publisher
権利情報
This research was originally published in the Journal of Biological Chemistry. H. Takahashi, T. Kumagai, K. Kitani, M. Mori, Y. Matoba, and M. Sugiyama. Cloning and Characterization of a Streptomyces Single Module Type Non-ribosomal Peptide Synthetase Catalyzing a Blue Pigment Synthesis. J. Biol. Chem. 2007; 282:pp.9073-9081. © the American Society for Biochemistry and Molecular Biology
関連情報URL
部局名
医歯薬学総合研究科



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