文献类型：期刊文献

英文题名：Intracellular Environment Improvement of Mycobacterium neoaurum for Enhancing Androst-1,4-Diene-3,17-Dione Production by Manipulating NADH and Reactive Oxygen Species Levels

作者：Shao, Minglong[1];Zhao, Youxi[2];Liu, Yu[1];Yang, Taowei[1];Xu, Meijuan[1];Zhang, Xian[1];Rao, Zhiming[1]

第一作者：Shao, Minglong

通讯作者：Rao, ZM[1]

机构：[1]Jiangnan Univ, Sch Biotechnol, Minist Educ, Key Lab Ind Biotechnol, Wuxi 214122, Jiangsu, Peoples R China;[2]Beijing Union Univ, Coll Biochem Engn, Beijing Key Lab BiomassWaste Resource Utilizat, Beijing 10023, Peoples R China

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

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

年份：2019

卷号：24

期号：21

外文期刊名：MOLECULES

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000498055500030)】；

基金：This research was funded by the National Natural Science Foundation of China (31700041 and 31570085), the National Key Research and Development Program of China (2018YFA0900304), National First-Class Discipline Program of Light Industry Technology and Engineering (LITE 2018-06) and the 111 Project (111-2-06).

语种：英文

外文关键词：androst-1,4-diene-3,17-dione; intracellular environment; NADH oxidase; catalase; Mycobacterium neoaurum

摘要：As one of the most significant steroid hormone precursors, androst-1,4-diene-3,17-dione (ADD) could be used to synthesize many valuable hormone drugs. The microbial transformation of sterols to ADD has received extensive attention in recent years. In a previous study, Mycobacterium neoaurum JC-12 was isolated and converted sterols to the major product, ADD. In this work, we enhanced ADD yield by improving the cell intracellular environment. First, we introduced a nicotinamide adenine dinucleotide (NADH) oxidase from Bacillus subtilis to balance the intracellular NAD(+) availability in order to strengthen the ADD yield. Then, the catalase gene from M. neoaurum was also over-expressed to simultaneously scavenge the generated H2O2 and eliminate its toxic effects on cell growth and sterol transformation. Finally, using a 5 L fermentor, the recombinant strain JC-12(yodC-katA) produced 9.66 g/L ADD, which increased by 80% when compared with the parent strain. This work shows a promising way to increase the sterol transformation efficiency by regulating the intracellular environment.