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Domain Characterization of Cyclosporin Regio-Specific Hydroxylases in Rare Actinomycetes
Woo, Min-Woo1; Lee, Bo-Ram1; Nah, Hee-Ju1; Choi, Si-Sun1; Li, Shengying2; Kim, Eung-Soo1
2015-10-01
发表期刊JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY
卷号25期号:10页码:1634-1639
摘要Cytochrome P450 hydroxylase (CYP) in actinomycetes plays an important role in the biosynthesis and bioconversion of various secondary metabolites. Two unique CYPs named CYP-sb21 and CYP-pa1, which were identified from Sebekia benihana and Pseudonocardia autotrophica, respectively, were proven to transfer a hydroxyl group at the 4th or 9th N-methyl leucine position of immunosuppressive agent cyclosporin A (CsA). Interestingly, these two homologous CYPs showed different CsA regio-selectivities. CYP-sb21 exhibited preferential hydroxylation activity at the 4th position over the 9th position, whereas CYP-pa1 showed the opposite preference. To narrow down the CYP domain critical for CsA regio-selectivity, each CYP was divided into four domains, and each domain was swapped with its counterpart from the other CYP. A total of 18 hybrid CYPs were then individually tested for CsA regioselectivity. Although most of the hybrid CYPs failed to exhibit a significant change in regioselectivity in the context of CsA hydroxylation, hybrid CYP-pa1 swapped with the second domain of CYP-sb21 showed a higher preference for the 9th position. Moreover, hybrid CYP-sb21 containing seven amino acids from the 2nd domain of CYP-pa1 showed higher preference for the 4th position. These results imply that the 2nd domain of CsA-specific CYP plays a critical role in CsA regio-selectivity, thereby setting the stage for biotechnological application of CsA regio-selective hydroxylation.
文章类型Article
关键词Cytochrome P450 Cyclosporin a Regio-selectivity Actinomycetes
WOS标题词Science & Technology ; Life Sciences & Biomedicine
DOI10.4014/jmb.1505.05034
关键词[WOS]CYTOCHROME-P450 COMPLEMENT CYPOME ; SEBEKIA-BENIHANA ; FERREDOXIN REDUCTASE ; STREPTOMYCES ; GENE ; DISRUPTION ; CONVERSION ; BINDING
收录类别SCI
语种英语
WOS研究方向Biotechnology & Applied Microbiology ; Microbiology
项目资助者National Research Foundation of Korea (NRF) - Korea government (MSIP)(NRF-2014R1A2A1A11052236)
WOS类目Biotechnology & Applied Microbiology ; Microbiology
WOS记录号WOS:000365443800008
引用统计
文献类型期刊论文
条目标识符http://ir.qibebt.ac.cn/handle/337004/6799
专题酶工程研究组
作者单位1.Inha Univ, Dept Biol Engn, Inchon 402751, South Korea
2.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Qingdao 266101, Shandong, Peoples R China
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Woo, Min-Woo,Lee, Bo-Ram,Nah, Hee-Ju,et al. Domain Characterization of Cyclosporin Regio-Specific Hydroxylases in Rare Actinomycetes[J]. JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY,2015,25(10):1634-1639.
APA Woo, Min-Woo,Lee, Bo-Ram,Nah, Hee-Ju,Choi, Si-Sun,Li, Shengying,&Kim, Eung-Soo.(2015).Domain Characterization of Cyclosporin Regio-Specific Hydroxylases in Rare Actinomycetes.JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY,25(10),1634-1639.
MLA Woo, Min-Woo,et al."Domain Characterization of Cyclosporin Regio-Specific Hydroxylases in Rare Actinomycetes".JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY 25.10(2015):1634-1639.
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