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- Title
Shaping the lipid composition of bacterial membranes for membrane protein production.
- Authors
Kanonenberg, Kerstin; Royes, Jorge; Kedrov, Alexej; Poschmann, Gereon; Angius, Federica; Solgadi, Audrey; Spitz, Olivia; Kleinschrodt, Diana; Stühler, Kai; Miroux, Bruno; Schmitt, Lutz
- Abstract
Background: 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. Results: 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). Conclusions: 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.
- Subjects
MEMBRANE proteins; BACTERIAL cell walls; ATP-binding cassette transporters; COMPLEMENTATION (Genetics); LIPIDS
- Publication
Microbial Cell Factories, 2019, Vol 18, Issue 1, pN.PAG
- ISSN
1475-2859
- Publication type
Article
- DOI
10.1186/s12934-019-1182-1