In vitro reconstitution of the cyclosporine specific P450 hydroxylases using heterologous redox partner proteins

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• 作者：Yue Sun ; Li Ma ; Dongfei Han ; Lei Du…
• 关键词：P450 hydroxylase ; Cyclosporin A ; Hair ; stimulating agent ; Redox partner proteins ; Synechococcus elongatus
• 刊名：Journal of Industrial Microbiology & Biotechnology
• 出版年：2017
• 出版时间：February 2017
• 年：2017
• 卷：44
• 期：2
• 页码：161-166
• 全文大小：
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Microbiology; Biochemistry, general; Inorganic Chemistry; Genetic Engineering; Biotechnology; Bioinformatics;
• 出版者：Springer Berlin Heidelberg
• ISSN：1476-5535
• 卷排序：44

文摘

The cytochrome P450 enzymes (CYPs) CYP-sb21 from Sebekia benihana and CYP-pa1 from Pseudonocardia autotrophica are able to hydroxylate the immunosuppressant cyclosporin A (CsA) in a regioselective manner, giving rise to the production of two hair-stimulating agents (with dramatically attenuated immunosuppressant activity), γ-hydroxy-N-methyl-l-Leu4-CsA (CsA-4-OH) and γ-hydroxy-N-methyl-l-Leu9-CsA (CsA-9-OH). Recently, the in vitro activity of CYP-sb21 was identified using several surrogate redox partner proteins. Herein, we reconstituted the in vitro activity of CYP-pa1 for the first time via a similar strategy. Moreover, the supporting activities of a set of ferredoxin (Fdx)/ferredoxin reductase (FdR) pairs from the cyanobacterium Synechococcus elongatus PCC 7942 were comparatively analyzed to identify the optimal redox systems for these two CsA hydroxylases. The results suggest the great value of cyanobacterial redox partner proteins for both academic research and industrial application of P450 biocatalysts.