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

• Published in: Frontiers in bioengineering and biotechnology Vol. 7; p. 408
• Main Authors: Kassab, Elias, Mehlmer, Norbert, Brueck, Thomas
• Format: Journal Article
• Language: English
• Published: Switzerland Frontiers Media S.A 10.12.2019
• Subjects: Arabidopsis thaliana; Bioengineering and Biotechnology; Escherichia coli; fatty acid biosynthesis; self-assembly GFP; VLCFA; Arabidopsis thaliana; self-assembly GFP; Escherichia coli; fatty acid biosynthesis; VLCFA
• ISSN: 2296-4185; 2296-4185
• DOI: 10.3389/fbioe.2019.00408

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, 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 ( ) enzymatic cascade in to enable unbranched VLCFA biosynthesis. The four 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).