Sun Yue, Ma Li, Han Dongfei, Du Lei, Qi Fengxia, Zhang Wei, Sun Jingran, Huang Shan, Kim Eung-Soo, Li Shengying
College of Chemical Engineering, Qingdao University of Science and Technology, Qingdao, Shandong, China.
Shandong Provincial Key Laboratory of Synthetic Biology, CAS Key Laboratory of Biofuels, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, Shandong, China.
J Ind Microbiol Biotechnol. 2017 Feb;44(2):161-166. doi: 10.1007/s10295-016-1875-y. Epub 2016 Nov 25.
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.
来自贝氏塞贝克菌的细胞色素P450酶（CYPs）CYP-sb21和来自自养假诺卡氏菌的CYP-pa1能够以区域选择性方式使免疫抑制剂环孢菌素A（CsA）羟基化，从而产生两种毛发生长促进剂（免疫抑制活性显著减弱），即γ-羟基-N-甲基-L-亮氨酸4-CsA（CsA-4-OH）和γ-羟基-N-甲基-L-亮氨酸9-CsA（CsA-9-OH）。最近，利用几种替代氧化还原伴侣蛋白鉴定了CYP-sb21的体外活性。在此，我们首次通过类似策略重建了CYP-pa1的体外活性。此外，对来自聚球藻PCC 7942的一组铁氧化还原蛋白（Fdx）/铁氧化还原蛋白还原酶（FdR）对的支持活性进行了比较分析，以确定这两种CsA羟化酶的最佳氧化还原系统。结果表明，蓝细菌氧化还原伴侣蛋白对于P450生物催化剂的学术研究和工业应用都具有重要价值。
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