Wang Sumeng, Jin Xin, Jiang Wei, Wang Qian, Qi Qingsheng, Liang Quanfeng
State Key Laboratory of Microbial Technology, Shandong University, Jinan, China.
Front Microbiol. 2022 May 11;13:902848. doi: 10.3389/fmicb.2022.902848. eCollection 2022.
3-Hydroxypropionic acid (3-HP) is a promising high value-added chemical. Acetyl-CoA carboxylase (Acc) is a vital rate-limiting step in 3-HP biosynthesis through the malonyl-CoA pathway. However, Acc toxicity in cells during growth blocks its ability to catalyze acetyl-CoA to malonyl-CoA. The balancing of Acc and malonyl-CoA reductase (MCR) expression is another an unexplored but key process in 3-HP production. To solve these problems, in the present study, we developed a method to mitigate Acc toxicity cell growth through Acc subunits (AccBC and DtsR1) expression adjustment. The results revealed that cell growth and 3-HP production can be accelerated through the adjustment of DtsR1 and AccBC expression. Subsequently, the balancing Acc and MCR expression was also employed for 3-HP production, the engineered strain achieved the highest titer of 6.8 g/L, with a high yield of 0.566 g/g glucose and productivity of 0.13 g/L/h, in shake-flask fermentation through the malonyl-CoA pathway. Likewise, the engineered strain also had the highest productivity (1.03 g/L/h) as well as a high yield (0.246 g/g glucose) and titer (up to 38.13 g/L) in fed-batch fermentation, constituting the most efficient strain for 3-HP production through the malonyl-CoA pathway using a cheap carbon source. This strategy might facilitate the production of other malonyl-CoA-derived chemical compounds in the future.
3-羟基丙酸(3-HP)是一种很有前景的高附加值化学品。乙酰辅酶A羧化酶(Acc)是通过丙二酰辅酶A途径进行3-HP生物合成的关键限速步骤。然而,生长过程中细胞内Acc的毒性阻碍了其将乙酰辅酶A催化为丙二酰辅酶A的能力。Acc和丙二酰辅酶A还原酶(MCR)表达的平衡是3-HP生产中另一个尚未探索但关键的过程。为了解决这些问题,在本研究中,我们开发了一种通过调节Acc亚基(AccBC和DtsR1)的表达来减轻Acc对细胞生长毒性的方法。结果表明,通过调节DtsR1和AccBC的表达可以加速细胞生长和3-HP的产生。随后,还利用Acc和MCR表达的平衡来生产3-HP,在摇瓶发酵中,通过丙二酰辅酶A途径,工程菌株达到了6.8 g/L的最高滴度,葡萄糖产率高达0.566 g/g,生产力为0.13 g/L/h。同样,在补料分批发酵中,该工程菌株也具有最高的生产力(1.03 g/L/h)以及高产量(0.246 g/g葡萄糖)和滴度(高达38.13 g/L),是使用廉价碳源通过丙二酰辅酶A途径生产3-HP的最有效菌株。该策略可能会在未来促进其他丙二酰辅酶A衍生化合物的生产。