(228az) Pathway Compartmentalization in Peroxisome of Saccharomyces Cerevisiae to Produce Versatile Medium Chain Fatty Alcohols

Fatty alcohols are value-added chemicals and important components of a variety of industries, which have a >3 billion-dollar global market annually. Long chain fatty alcohols (>C12) are mainly used in surfactants, lubricants, detergents, pharmaceuticals and cosmetics while medium chain fatty alcohols (C6-C12) could be used as diesel-like biofuels. Microbial production of fatty alcohols from renewable feedstock stands as a promising strategy to enable sustainable supply of this important chemical. In this study, we report, for the first time, that medium chain fatty alcohols could be produced in Saccharomyces cerevisiae via targeted expression of a fatty acyl-CoA reductase (TaFAR or AdFAR) in the peroxisome. By tagging FAR enzymes with peroxisomal targeting signal peptides, the FAR could be compartmentalized into the matrix of the peroxisome to hijack the medium chain fatty acyl-CoA generated from the beta-oxidation pathway and convert them to versatile medium chain fatty alcohols (C10&C12). The overexpression of genes encoding PEX7 and acetyl-CoA carboxylase further improved fatty alcohol production by 1.4-fold. Fatty alcohols were produced at 1.3 g/L in fed-batch fermentation under nitrogen limited condition, including 6.9% 1-decanol, 27.5% 1-dodecanol, 2.9% 1-tetradecanol and 62.7% 1-hexadecanol. This work revealed that peroxisome could be engineered as a compartmentalized cell factory for producing versatile fatty acid-derived chemicals in S. cerevisiae.