(311d) Rational Engineering of Clostridium for Efficient Production of Renewable Fatty Acid Esters

Authors: 
Wang, Y., Auburn University
Feng, J., Auburn University
Zhang, J., Auburn University
Borovok, I., Tel Aviv University
Cao, M., University of Illinois at Urbana-Champaign
Shao, Z., Iowa State University
Production of renewable chemicals through biological routes is considered as an urgent solution for fossil energy crisis. However, endproduct toxicity inhibits microbial host 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, 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. Specifically, the butyl acetate production we obtained is 2400-fold higher than the highest level that has been previously reported in microbial hosts. Besides, some of our engineered strains demonstrated great potential for producing other esters as well. Our principles of selecting the most appropriate host for specific bioproduction and engineering microbial hosts 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.