Shaping the lipid composition of bacterial membranes for membrane protein production

• Published in: Microbial cell factories Vol. 18; no. 1; p. 131
• Main Authors: Kanonenberg, Kerstin, Royes, Jorge, Kedrov, Alexej, Poschmann, Gereon, Angius, Federica, Solgadi, Audrey, Spitz, Olivia, Kleinschrodt, Diana, Stühler, Kai, Miroux, Bruno, Schmitt, Lutz
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 10.08.2019; BioMed Central; BMC
• Subjects: ATP-Binding Cassette Transporters - chemistry; ATP-Binding Cassette Transporters - genetics; Biotechnology; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; Escherichia coli Proteins - biosynthesis; Escherichia coli Proteins - chemistry; Escherichia coli Proteins - genetics; Fatty acids; Gene expression; Life Sciences; Lipidome; Lipids - chemistry; Membrane engineering; Membrane protein; Membrane proteins; Membrane Proteins - biosynthesis; Membrane Proteins - genetics; Membrane Transport Proteins - chemistry; Membrane Transport Proteins - genetics; Metabolic Engineering; Proteins; SEC Translocation Channels - chemistry; SEC Translocation Channels - genetics; Solubilization; Strain engineering; Membrane engineering; Membrane protein; Lipidome; Solubilization; Strain engineering
• ISSN: 1475-2859; 1475-2859
• DOI: 10.1186/s12934-019-1182-1

The overexpression and purification of membrane proteins is a bottleneck in biotechnology and structural biology. E. coli remains the host of choice for membrane protein production. To date, most of the efforts have focused on genetically tuning of expression systems and shaping membrane composition to improve membrane protein production remained largely unexplored. In E. coli C41(DE3) strain, we deleted two transporters involved in fatty acid metabolism (OmpF and AcrB), which are also recalcitrant contaminants crystallizing even at low concentration. Engineered expression hosts presented an enhanced fitness and improved folding of target membrane proteins, which correlated with an altered membrane fluidity. We demonstrated the scope of this approach by overproducing several membrane proteins (4 different ABC transporters, YidC and SecYEG). In summary, E. coli membrane engineering unprecedentedly increases the quality and yield of membrane protein preparations. This strategy opens a new field for membrane protein production, complementary to gene expression tuning.