Construction of a novel expression vector in Pseudonocardia autotrophica and its application to efficient biotransformation of compactin to pravastatin, a specific HMG-CoA reductase inhibitor

Fujii, Yoshikazu; Norihisa, Koji; Fujii, Tadashi; Aritoku, Yasuhide; Kagawa, Yusuke; Sallam, Khalid Ibrahim; Johdo, Osamu; Arisawa, Akira; Tamura, Tomohiro

Construction of a novel expression vector in Pseudonocardia autotrophica and its application to efficient biotransformation of compactin to pravastatin, a specific HMG-CoA reductase inhibitor

Fujii, Yoshikazu; Norihisa, Koji; Fujii, Tadashi et al.

Biochemical and Biophysical Research Communications, 2011, 404(1), 511-516

Permalink : https://hdl.handle.net/2115/44922

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アクセス権	open access
URI	https://hdl.handle.net/2115/44922
タイトル	Construction of a novel expression vector in Pseudonocardia autotrophica and its application to efficient biotransformation of compactin to pravastatin, a specific HMG-CoA reductase inhibitor
著者	Fujii, Yoshikazu ; Norihisa, Koji ; Fujii, Tadashi ; Aritoku, Yasuhide ; Kagawa, Yusuke ; Sallam, Khalid Ibrahim ; Johdo, Osamu ; Arisawa, Akira ; Tamura, Tomohiro
著者名 Fujii, Yoshikazu
著者名 Norihisa, Koji
著者名 Fujii, Tadashi
著者名 Aritoku, Yasuhide
著者名 Kagawa, Yusuke
著者名 Sallam, Khalid Ibrahim
著者名 Johdo, Osamu
著者名 Arisawa, Akira
e-Rad_Researcher 30357488 著者名 Tamura, Tomohiro
言語	英語
キーワード	Pseudonocardia autotrophica
	Pravastatin
	Rolling-circle type replication
	Acetone-inducible promoter
	Biotransformation
発行日	2011-01-07
出版者	Elsevier
収録物名	Biochemical and Biophysical Research Communications
巻(号)	404(1)
ページ	511-516
抄録	The novel plasmid vector (pTAOR4-Rev) suitable for gene expression in actinomycete strains of Pseudonocardia autotrophica was constructed from 2 P. autotrophica genetic elements, the novel replication origin and the acetone-inducible promoter. The replication origin was isolated from the endogenous plasmid of strain DSM 43082 and the acetone-inducible promoter was determined by analysis of the upstream region of an acetaldehyde dehydrogenase gene homologue in strain NBRC 12743. P. autotrophica strains transformed with pTAOR4-P450, carrying a gene for cytochrome P450 monooxygenase, expressed P450 from the acetone-inducible promoter, as verified by SDS-PAGE and spectral analysis. The biotransformation test of acetone-induced resting cells prepared from a strain of P. autotrophica carrying pTAOR4 that harbors a compactin (CP)-hydroxylating P450 gene revealed 3.3-fold increased production of pravastatin (PV), a drug for hypercholesterolemia. Biotransformation of CP by the same strain in batch culture yielded PV accumulation of 14.3 g/l after 100 hr. The expression vector pTAOR4-Rev and its function-enhancing derivatives provide a versatile approach to industrial biotransformation by Pseudonocardia strains, which can be good hosts for P450 monooxygenase expression.
資源タイプ	学術雑誌論文
出版タイプ	Accepted Manuscript
PISSN	0006-291X
関連情報 (isVersionOf)	出版社版DOI https://doi.org/10.1016/j.bbrc.2010.12.013