School of Biological Sciences and Technology, Chonnam National University, Kwangju, 500-757, Republic of Korea.
Arch Pharm Res. 2015 Jan;38(1):98-107. doi: 10.1007/s12272-014-0366-9. Epub 2014 Mar 24.
In this study, we examined the catalytic activity of CYP106A1 from the Bacillus megaterium American Type Culture Collection 14581 strain. The CYP106A1 gene was cloned from B. megaterium, heterologously expressed in Escherichia coli, and purified. Potential electron partners and possible bacterial CYP106A1 substrates were identified by examining the oxidative activity toward a set of steroids in the presence of several reductase systems. The activities of CYP106A1 in a reconstituted system could not be achieved using rat NADPH-P450 reductase or a putidaredoxin reductase-putidaredoxin pair. However, the spinach redox proteins, a ferredoxin reductase-ferredoxin pair, were found to be efficient redox partners for CYP106A1. CYP106A1 catalyzes the hydroxylation of a set of steroids including testosterone, progesterone, 17α-hydroxyprogesterone, 11-deoxycorticosterone, corticosterone, and 11-deoxycortisol to produce monohydroxylated products as the major metabolites. These results suggest that CYP106A1 would be useful for the bioconversion of steroid hormones to hydroxylated products that can be used for industrial applications.
在这项研究中，我们研究了来自巨大芽孢杆菌美国典型培养物保藏 14581 株的 CYP106A1 的催化活性。CYP106A1 基因从巨大芽孢杆菌中克隆，在大肠杆菌中异源表达并纯化。通过检查一系列甾体在几种还原酶系统存在下的氧化活性，确定了潜在的电子伙伴和可能的细菌 CYP106A1 底物。在重组系统中，CYP106A1 的活性不能使用大鼠 NADPH-P450 还原酶或 putidaredoxin 还原酶-putidaredoxin 对来实现。然而，菠菜氧化还原蛋白，即一种铁氧还蛋白还原酶-铁氧还蛋白对，被发现是 CYP106A1 的有效氧化还原伴侣。CYP106A1 催化包括睾酮、孕酮、17α-羟孕酮、11-脱氧皮质酮、皮质酮和 11-脱氧皮质醇在内的一组甾体的羟化作用，产生单羟化产物作为主要代谢物。这些结果表明，CYP106A1 将可用于甾体激素向可用于工业应用的羟化产物的生物转化。
