Ma Ruoshuang, Fang Haitian, Liu Huiyan, Pan Lin, Wang Hongyan, Zhang Heng
Ningxia Key Laboratory for Food Microbial-Applications Technology and Safety Control, School of Agriculture, Ningxia University, Yinchuan, 750021, China.
Biotechnol Lett. 2021 Jun;43(6):1211-1219. doi: 10.1007/s10529-021-03103-3. Epub 2021 Mar 1.
Cytidine is an important raw material for nucleic acid health food and genetic engineering research. In recent years, it has shown irreplaceable effects in anti-virus, anti-tumor, and AIDS drugs. Its biosynthetic pathway is complex and highly regulated. In this study, overexpression of uracil permease and a nucleoside transporter from Bacillus amyloliquefaciens related to cell membrane transport in Escherichia coli strain BG-08 was found to increase cytidine production in shake flask cultivation by 1.3-fold (0.91 ± 0.03 g/L) and 1.8-fold (1.26 ± 0.03 g/L) relative to that of the original strain (0.70 ± 0.03 g/L), respectively. Co-overexpression of uracil permease and a nucleoside transporter further increased cytidine yield by 2.7-fold (1.59 ± 0.05 g/L) compared with that of the original strain. These results indicate that the overexpressed uracil permease and nucleoside transporter can promote the accumulation of cytidine, and the two proteins play a synergistic role in the secretion of cytidine in Escherichia coli.
胞苷是核酸保健食品和基因工程研究的重要原料。近年来,它在抗病毒、抗肿瘤和艾滋病药物中显示出不可替代的作用。其生物合成途径复杂且受到高度调控。在本研究中,发现在大肠杆菌菌株BG - 08中过表达来自解淀粉芽孢杆菌的与细胞膜转运相关的尿嘧啶通透酶和核苷转运蛋白,相对于原始菌株(0.70±0.03 g/L),摇瓶培养中的胞苷产量分别提高了1.3倍(0.91±0.03 g/L)和1.8倍(1.26±0.03 g/L)。与原始菌株相比,尿嘧啶通透酶和核苷转运蛋白的共过表达使胞苷产量进一步提高了2.7倍(1.59±0.05 g/L)。这些结果表明,过表达的尿嘧啶通透酶和核苷转运蛋白可以促进胞苷的积累,并且这两种蛋白质在大肠杆菌中胞苷的分泌中发挥协同作用。