文献类型：期刊文献

英文题名：High-level living cell production of cytidine-5′-diphosphocholine in metabolically engineered yeast

作者：Ren, Yanna[1];Liu, Qi[1];Liu, Haifeng[2];Zhou, Xiangshan[2,3];Zhang, Yuanxing[4];Cai, Menghao[1]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]China Resources Angde Biotech Pharma Co Ltd, 78 E Jiao St, Liaocheng, Shandong, Peoples R China;[3]China Resources Biopharmaceut Co Ltd, 1301-84 Sightseeing Rd, Shenzhen, Peoples R China;[4]Shanghai Collaborat Innovat Ctr Biomfg, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2021

卷号：341

起止页码：129

外文期刊名：JOURNAL OF BIOTECHNOLOGY

收录：;EI(收录号：20214110997695);WOS:【SCI-EXPANDED(收录号:WOS:000712544600006)】；

基金：The authors are grateful for the support of this work by the National Key R&D Program of China (2020YFA0907800) , National Natural Science Foundation of China (31870073) , Shanghai Rising-Star Program, China (19QA1402700) , 111 Project of China (B18022) , Fundamental Research Funds for the Central Universities in China (22221818014) , Research Program of State Key Laboratory of Bioreactor Engineering. We thank Nantong QZU Bioscience & Biotechnology Co. LTD for providing the choline phosphate.

语种：英文

外文关键词：Living cell production; Metabolic engineering; Cytidine-5 '-diphosphocholine; Pichia pastoris

摘要：Industrial production of neuroprotective drug CDP-choline is accomplished via permeabilized or lysed cell biotransformation because of the inefficient penetration of substrates into intact cells. We previously proposed a novel one-step living cell method for CDP-choline production by engineered yeast, but obtained low titer and molar yield. This study develops a high-production strain with improved molar yield by metabolic engineering strategies. The selective markers previously integrated into host cell were recovered for facilitating genetic modification, which however resulted a strain with improved CDP-choline titer and molar yield to CMP. Knockout of 5'-NT or CDA in CMP sinking pathway but not APY in CTP sinking pathway further improved CDP-choline titer and molar yield to CMP. However, overexpression of seven enzymes in CTP synthetic pathway showed no positive functions. Finally, optimization of CMP and choline phosphate levels for the optimized recombinant strains achieved a high-level CDP-choline of similar to 30 g/L, which was enhanced by 400% compared to the previous work. Also, the molar yield of CDP-choline to CMP increased from 40% to 84.7%. The titer and molar yield are comparable to the reported permeabilized or lysed cell based biotransformation methods. It represents a novel and competitive paradigm for the potential industrial production of CDP-choline.