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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
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)
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Submitter: 田村 具博
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