文献类型：期刊文献

英文题名：A Novel 3-Phytosterone-9 alpha-Hydroxylase Oxygenation Component and Its Application in Bioconversion of 4-Androstene-3,17-Dione to 9 alpha-Hydroxy-4-Androstene-3,17-Dione Coupling with A NADH Regeneration Formate Dehydrogenase

作者：Zhang, Xian[1];Zhu, Manchi[1];Han, Rumeng[1];Zhao, Youxi[2];Chen, Kewei[3];Qian, Kai[4];Shao, Minglong[1];Yang, Taowei[1];Xu, Meijuan[1];Xu, Jianzhong[1];Rao, Zhiming[1]

第一作者：Zhang, Xian

通讯作者：Xu, JZ[1];Rao, ZM[1]

机构：[1]Jiangnan Univ, Sch Biotechnol, Key Lab Ind Biotechnol, Minist Educ, 1800 Liu Ave, Wuxi 214122, Jiangsu, Peoples R China;[2]Beijing Union Univ, Biochem Engn Coll, Beijing 100023, Peoples R China;[3]Jiangnan Univ, Sch Food Sci & Technol, 1800 Liu Ave, Wuxi 214122, Jiangsu, Peoples R China;[4]Yichun Univ, Sch Med, Yichun 336000, Jiangxi, Peoples R China

第一机构：Jiangnan Univ, Sch Biotechnol, Key Lab Ind Biotechnol, Minist Educ, 1800 Liu Ave, Wuxi 214122, Jiangsu, Peoples R China

通讯机构：[1]corresponding author), Jiangnan Univ, Sch Biotechnol, Key Lab Ind Biotechnol, Minist Educ, 1800 Liu Ave, Wuxi 214122, Jiangsu, Peoples R China.

年份：2019

卷号：24

期号：14

外文期刊名：MOLECULES

收录：;Scopus(收录号：2-s2.0-85068911173);WOS:【SCI-EXPANDED(收录号:WOS:000482303000017)】；

基金：This work was supported by the National Natural Science Foundation of China (31500065, 21778024, 31870066), the National Key Research and Development Program of China (2018YFA0900304), national first-class discipline program of Light Industry Technology and Engineering (LITE2018-06), key research and development program of Ningxia hui autonomous region (2017BY069, 2019BCH01002), the science and technology innovation team foundation of Ningxia hui autonomous region (KJT2017001), the Top-Notch Academic Programs Project of Jiangsu Higher Education Institutions, 111 Project, Program of the Key Laboratory of Industrial Biotechnology, Ministry of Education, China (KLIB-KF201902), the General Project of Beijing Municipal Education Commission (SQKM201311417004), the Opening Foundation of Beijing Key Laboratory of BiomassWaste Resource Utilization, Jiangsu province "Collaborative Innovation Center for Advanced Industrial Fermentation" industry development program.

语种：英文

外文关键词：9 alpha-Hydroxy-4-androstene-3; 17-dione; 3-phytosterone-9 alpha-hydroxylase; hydroxylation; NADH regeneration

摘要：9 alpha-Hydroxy-4-androstene-3,17-dione (9-OH-AD) is one of the significant intermediates for the preparation of beta-methasone, dexamethasone, and other steroids. In general, the key enzyme that enables the biotransformation of 4-androstene-3,17-dione (AD) to 9-OH-AD is 3-phytosterone-9 alpha-hydroxylase (KSH), which consists of two components: a terminal oxygenase (KshA) and ferredoxin reductase (KshB). The reaction is carried out with the concomitant oxidation of NADH to NAD(+). In this study, the more efficient 3-phytosterone-9 alpha-hydroxylase oxygenase (KshC) from the Mycobacterium sp. strain VKM Ac-1817D was confirmed and compared with reported KshA. To evaluate the function of KshC on the bioconversion of AD to 9-OH-AD, the characterization of KshC and the compounded system of KshB, KshC, and NADH was constructed. The optimum ratio of KSH oxygenase to reductase content was 1.5:1. An NADH regeneration system was designed by introducing a formate dehydrogenase, further confirming that a more economical process for biological transformation from AD to 9-OH-AD was established. A total of 7.78 g of 9-OH-AD per liter was achieved through a fed-batch process with a 92.11% conversion rate (mol/mol). This enzyme-mediated hydroxylation method provides an environmentally friendly and economical strategy for the production of 9-OH-AD.