Endocytosing Escherichia coil as a Whole-Cell Biocatalyst of Fatty Acids

Abstract

Whole cell biocatalysts can be used to convert fatty acids into various value-added products. However, fatty acid transport across cellular membranes into the cytosol of microbial cells limits substrate availability and impairs membrane integrity, which in turn decreases cell viability and bioconversion activity. Because these problems are associated with the mechanism of fatty acid transport through membranes, a whole cell biocatalyst that can form caveolae-like structures was generated to promote substrate endocytosis. Caveolin-1 (CAVI) expression in Escherichia coli increased both the fatty acid transport rate and intracellular fatty acid concentrations via endocytosis of the supplemented substrate. Furthermore, fatty-acid endocytosis alleviated substrate cytotoxicity in E. coli. These traits attributed to bacterial endocytosis resulted in dramatically elevated biotransformation efficiencies in fed-batch and cell recycle reaction systems when caveolae-forming E. coli was used for the bioconversion of ricinoleic acid (12-hydroxyoctadec-9-enoic acid) to (Z)-11-(heptanoyloxy) undec-9-enoic acid. We propose that CAVI-mediated endocytosing E. coli represents a versatile tool for the biotransformation of hydrophobic substrates.