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Lu, Xiaoyun; Liu, Yuwan; Yang, Yiqun; Wang, Shanshan; Wang, Qian; Wang, Xiya; Yan, Zhihui; Cheng, Jian; Liu, Cui; Yang, Xue; Luo, Hao; Yang, Sheng; Gou, Junran; Ye, Luzhen; Lu, Lina; Zhang, Zhidan; Guo, Yu; Nie, Yan; Lin, Jianping; Li, Sheng; Tian, Chaoguang; Cai, Tao; Zhuo, Bingzhao; Ma, Hongwu; Wang, Wen; Ma, Yanhe; Liu, Yongjun; Li, Yin; Jiang, Huifeng
Nature communications, 03/2019, Volume: 10, Issue: 1Journal Article
Acetyl-CoA is a fundamental metabolite for all life on Earth, and is also a key starting point for the biosynthesis of a variety of industrial chemicals and natural products. Here we design and construct a Synthetic Acetyl-CoA (SACA) pathway by repurposing glycolaldehyde synthase and acetyl-phosphate synthase. First, we design and engineer glycolaldehyde synthase to improve catalytic activity more than 70-fold, to condense two molecules of formaldehyde into one glycolaldehyde. Second, we repurpose a phosphoketolase to convert glycolaldehyde into acetyl-phosphate. We demonstrated the feasibility of the SACA pathway in vitro, achieving a carbon yield ~50%, and confirmed the SACA pathway by C-labeled metabolites. Finally, the SACA pathway was verified by cell growth using glycolaldehyde, formaldehyde and methanol as supplemental carbon source. The SACA pathway is proved to be the shortest, ATP-independent, carbon-conserving and oxygen-insensitive pathway for acetyl-CoA biosynthesis, opening possibilities for producing acetyl-CoA-derived chemicals from one-carbon resources in the future.
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