Functional analysis of a prenyltransferase gene (paxD) in the paxilline biosynthetic gene cluster

Liu, Chengwei; Noike, Motoyoshi; Minami, Atsushi; Oikawa, Hideaki; Dairi, Tohru

Functional analysis of a prenyltransferase gene (paxD) in the paxilline biosynthetic gene cluster

Liu, Chengwei; Noike, Motoyoshi; Minami, Atsushi et al.

Applied microbiology and biotechnology, 2014, 98(1), 199-206

Permalink : https://hdl.handle.net/2115/57639

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アクセス権	open access
URI	https://hdl.handle.net/2115/57639
タイトル	Functional analysis of a prenyltransferase gene (paxD) in the paxilline biosynthetic gene cluster
著者	Liu, Chengwei ; Noike, Motoyoshi ; Minami, Atsushi ; Oikawa, Hideaki ; Dairi, Tohru
著者名 Liu, Chengwei
著者名 Noike, Motoyoshi
著者名 Minami, Atsushi
著者名 Oikawa, Hideaki
e-Rad_Researcher 70264679 著者名 Dairi, Tohru
言語	英語
キーワード	Fungus
	Penicillium paxilli
	Indole-diterpene
	Paxilline
	Prenyltransferase
	PaxD
発行日	2014-01
出版者	Springer
収録物名	Applied microbiology and biotechnology
巻(号)	98(1)
ページ	199-206
抄録	Paxilline is an indole-diterpene produced by Penicillium paxilli. Six genes (paxB, C, G, M, P, and Q) in paxilline biosynthetic gene cluster were previously shown to be responsible for paxilline biosynthesis. In this study, we have characterized paxD, which is located next to paxQ and has weak similarities to fungal dimethylallyl tryptophan synthase genes. PaxD was overexpressed in Escherichia coli and the purified enzyme was used for in vitro analysis. When paxilline and dimethylallyl diphosphate were used as substrates, one major and one minor product, which were identified as di-prenyl paxilline and mono-prenyl paxilline by liquid chromatography-mass spectrometry analysis, were formed. The structure of the major product was determined to be 21,22-diprenylated paxilline, showing that PaxD catalyzed the successive di-prenylation. Traces of both products were detected in culture broth of P. paxilli by liquid chromatography-mass spectrometry analysis. The enzyme is likely to be a dimer and required no divalent cations. The optimum pH and temperature were 8.0 and 37 A degrees C, respectively. The Km values were calculated as 106.4 A +/- 5.4 mu M for paxilline and 0.57 A +/- 0.02 mu M for DMAPP with a kcat of 0.97 A +/- 0.01/s.
資源タイプ	学術雑誌論文
出版タイプ	Accepted Manuscript
PISSN	0175-7598
関連情報	PMID 23525886
関連情報 (isVersionOf)	出版社版DOI https://doi.org/10.1007/s00253-013-4834-9