Cloning and Characterization of a Streptomyces Single Module Type Non-ribosomal Peptide Synthetase Catalyzing a Blue Pigment Synthesis
この文献の参照には次のURLをご利用ください : https://ir.lib.hiroshima-u.ac.jp/00048634
ID | 48634 |
本文ファイル | |
著者 |
Takahashi, Hitoshi
大学院医歯薬学総合研究科
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
|
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
|
巻 | 282巻
|
号 | 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
|