文献类型：期刊文献

英文题名：Secretory expression of cyclohexanone monooxygenase by methylotrophic yeast for efficient omeprazole sulfide bio-oxidation

作者：Li, Ya-Jing[1];Zheng, Yu-Cong[1];Geng, Qiang[1];Liu, Feng[1];Zhang, Zhi-Jun[1,2];Xu, Jian-He[1,2];Yu, Hui-Lei[1,2]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol, Shanghai Collaborat Innovat Ctr Biomfg Technol, Shanghai 200237, Peoples R China

年份：2021

卷号：8

期号：1

外文期刊名：BIORESOURCES AND BIOPROCESSING

收录：;EI(收录号：20224513084543);WOS:【SCI-EXPANDED(收录号:WOS:000690897200001)】；

基金：This work was financially supported by the National Natural Science Foundation of China (21922804), the National Key Research and Development Program of China (2019YFA09005000), Program of Shanghai Academic Research Leader (21XD1400800) and the Fundamental Research Funds for the Central Universities (22221818014).

语种：英文

外文关键词：Cyclohexanone monooxygenase; Omeprazole sulfoxide; Pichia pastoris; Secretory expression; Asymmetric oxidation

摘要：Prochiral pyrmetazole can be asymmetrically oxidized into (S)-omeprazole, a proton pump inhibitor that is used to treat gastroesophageal reflux, by an engineered cyclohexanone monooxygenase (CHMOAcineto-Mut) that has high stereoselectivity. CHMOAcineto-Mut is produced by heterologous expression in Escherichia coli, where it is expressed intracellularly. Thus, isolating this useful biocatalyst requires tedious cell disruption and subsequent purification, which hinders its use for industrial purposes. Here, we report the extracellular production of CHMOAcineto-Mut by a methylotrophic yeast, Pichia pastoris, for the first time. The recombinant CHMOAcineto-Mut expressed by P. pastoris showed a higher flavin occupation rate than that produced by E. coli, and this was accompanied by a 3.2-fold increase in catalytic efficiency. At a cell density of 150 g/L cell dry weight, we achieved a recombinant CHMOAcineto-Mut production rate of 1,700 U/L, representing approximately 85% of the total protein secreted into the fermentation broth. By directly employing the pH adjusted supernatant as a biocatalyst, we were able to almost completely transform 10 g/L of pyrmetazole into the corresponding (S)-sulfoxide, with > 99% enantiomeric excess.