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Renewable Fatty Acid Ester Production in Clostridium

By Jun Feng, Jie Zhang, Yiming Feng, Pixiang Wang, Pablo Jiménez-Bonilla, Yanyan Gu, Junping Zhou, Zhong-Tian Zhang, Mingfeng Cao, Zengyi Shao, Ilya Borovok, Haibo Huang, Yi Wang

Posted 30 Mar 2020
bioRxiv DOI: 10.1101/2020.03.29.014746

Production of renewable chemicals through biological routes is considered as an urgent solution for fossil energy crisis. However, endproduct toxicity inhibits microbial performance and is a key bottleneck for biochemical production. To address this challenge, here we report an example of biosynthesis of high-value and easy-recoverable derivatives to alleviate endproduct toxicity and enhance bioproduction efficiency. By leveraging the natural pathways in solventogenic clostridia for co-producing acyl-CoAs, acids and alcohols as precursors, through rational screening for host strains and enzymes, systematic metabolic engineering— including rational organization of ester-synthesizing enzymes inside of the cell, and elimination of putative prophages, we developed strains that can produce 20.3 g/L butyl acetate and 1.6 g/L butyl butyrate respectively, which were both the unprecedented levels in microbial hosts. Techno-economic analysis indicated a production cost of $986 per metric tonne for butyl acetate production from corn stover comparing to the market price of $1,200-1,400 per metric tonne of butyl acetate, suggesting the economic competitiveness of our developed bioprocess. Our principles of selecting the most appropriate host for specific bioproduction and engineering microbial chassis to produce high-value and easy-separable endproducts are highly applicable to other bioprocesses, and could lead to breakthroughs in biofuel/biochemical production and general bioeconomy. ### Competing Interest Statement Auburn University has filed a patent application covering the work described in this article. The application names Y.W. and J.F. as inventors.

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