文献类型：期刊文献

英文题名：Fungal statin pump protein improves monacolin J efflux and regulates its production in Komagataella phaffii

作者：Bai, Chenxiao[1];Liu, Yiqi[1];Chen, Xinjie[1];Qian, Zhilan[1];Liu, Haifeng[2];Zhou, Xiangshan[1,2];Zhang, Yuanxing[1,3];Cai, Menghao[1]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]China Resources Angde Biotech Pharmaceut Co Ltd, 78 E Jiao St, Liaocheng, Shandong, Peoples R China;[3]Shanghai Collaborat Innovat Ctr Biomfg, Shanghai 200237, Peoples R China

年份：2020

卷号：7

期号：1

外文期刊名：BIORESOURCES AND BIOPROCESSING

收录：;EI(收录号：20224513087557);WOS:【SCI-EXPANDED(收录号:WOS:000543332100001)】；

基金：This work was supported by Fundamental Research Funds for the Shanghai Science and Technology Innovation Action Plan (Grant Number 17JC1402400) and the Shanghai Rising-Star Program, China (Grant Number 19QA1402700), the 111 Project of China (Grant Number B18022), the Fundamental Research Funds for the Central Universities, China (Grant Number JKF012016019, 22221818014) and Research Program of State Key Laboratory of Bioreactor Engineering.

语种：英文

外文关键词：Statin pump protein; Komagataella phaffii; Monacolin J; Metabolite efflux

摘要：Background Monacolin J (MJ) is a key intermediate for the synthesis of cholesterol-lowering drug simvastatin. Current industrial production of MJ involves complicated chemical hydrolysis of microbial fermented lovastatin. Recently, heterologous production of MJ has been achieved in yeast and bacteria, but the resulting metabolic stress and excessive accumulation of the compound adversely affect cell activity. Results Five genes,tapA, stapA, slovI, smokIand smlcE, coding for fungal statin pump proteins were expressed in an MJ producing yeast strain,Komagataella phaffiiJ#9. Overexpression of these genes facilitated MJ production. Among them,tapAfromAspergillus terreushighly improved MJ production and led to a titer increase of 108%. Exogenous MJ feeding study on an MJ non-producing strain GS-P-GAP-TapA was then performed, and the results illustrated tough entry of MJ into cells and possible efflux action of TapA. Further, intracellular and extracellular MJ levels of J#9 and J#9-TapA were analyzed. The extracellular MJ level of J#9-TapA increased faster, but its intracellular MJ percentage kept lower as compared to J#9. The results proved that TapA effectively excreted MJ from cells. Then functions of TapA were evaluated in a high-production bioreactor fermentation. Differently, TapA expression caused a low MJ titer but high intracellular MJ accumulation in J#9-TapA compared with J#9. Conclusions Statin pump proteins improved MJ production inK. phaffiiin a shake flask. Exogenous MJ feeding and endogenous MJ producing experiments demonstrated the efflux function of TapA. TapA improved MJ production at low MJ levels in a shake flask, but decreased it at high MJ levels in a bioreactor. This finding is useful for statin pump improvement and metabolic engineering for statin bioproduction.