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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

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Title: 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
Authors: Fujii, Yoshikazu Browse this author
Norihisa, Koji Browse this author
Fujii, Tadashi Browse this author
Aritoku, Yasuhide Browse this author
Kagawa, Yusuke Browse this author
Sallam, Khalid Ibrahim Browse this author
Johdo, Osamu Browse this author
Arisawa, Akira Browse this author
Tamura, Tomohiro Browse this author →KAKEN DB
Keywords: Pseudonocardia autotrophica
Pravastatin
Rolling-circle type replication
Acetone-inducible promoter
Biotransformation
Issue Date: 7-Jan-2011
Publisher: Elsevier
Journal Title: Biochemical and Biophysical Research Communications
Volume: 404
Issue: 1
Start Page: 511
End Page: 516
Publisher DOI: 10.1016/j.bbrc.2010.12.013
Abstract: 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.
Type: article (author version)
URI: http://hdl.handle.net/2115/44922
Appears in Collections:農学院・農学研究院 (Graduate School of Agriculture / Faculty of Agriculture) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 田村 具博

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