AUTHOR=Kassab Elias , Mehlmer Norbert , Brueck Thomas 

TITLE=GFP Scaffold-Based Engineering for the Production of Unbranched Very Long Chain Fatty Acids in Escherichia coli With Oleic Acid and Cerulenin Supplementation

JOURNAL=Frontiers in Bioengineering and Biotechnology

VOLUME=7

YEAR=2019

URL=https://www.frontiersin.org/journals/bioengineering-and-biotechnology/articles/10.3389/fbioe.2019.00408

DOI=10.3389/fbioe.2019.00408

ISSN=2296-4185

ABSTRACT=<p>Currently, very long chain fatty acids (VLCFAs) for oleochemical, pharmaceutical, cosmetic, or food applications are extracted from plant or marine organism resources, which is associated with a negative environmental impact. Therefore, there is an industrial demand to develop sustainable, microbial resources. Due to its ease of genetic modification and well-characterized metabolism, <italic>Escherichia coli</italic> has established itself as a model organism to study and tailor microbial fatty acid biosynthesis using a concerted genetic engineering approach. In this study, we systematically implemented a plant-derived (<italic>Arabidopsis thaliana</italic>) enzymatic cascade in <italic>Escherichia coli</italic> to enable unbranched VLCFA biosynthesis. The four <italic>Arabidopsis thaliana</italic> membrane-bound VLCFA enzymes were expressed using a synthetic expression cassette. To facilitate enzyme solubilization and interaction of the synthetic VLCFA synthase complex, we applied a self-assembly GFP scaffold. In order to initiate VLCFA biosynthesis, external oleic acid and cerulenin were supplemented to cultures. In this context, we detected the generation of arachidic (20:0), cis-11-eicosenoic (20:1) and cis-13-eicosenoic acid (20:1).</p>