Anaerobic Conditioning of E. coli Cell Lysate for Enhanced In Vitro Protein Synthesis.

ABSTRACT: Cell-free protein expression (CFPS) from E. coli cell lysate is an established chemical biology technique. Common efforts to improve synthesis capacity, such as strain engineering and process improvements, have overlooked the opportunity to increase productivity by reducing the dependence on limited, dissolved oxygen. Here we demonstrate conditioning E. coli cells for anaerobic respiration which increases the initial protein expression rate up to 4-fold and increases titer by 50% as compared to traditional aerobic cell lysate when using sfGFP as a reporter protein in CFPS reactions run at atmospheric conditions. This enhancement is even more significant when run in an oxygen-depleted environment, where anaerobic respiration preconditioned cells increase yield when supplemente

SUBMITTER: Tamiev BD 

PROVIDER: S-EPMC8168642 | biostudies-literature | 2021 Apr

REPOSITORIES: biostudies-literature