CRISPRi enables fast growth followed by stable aerobic pyruvate formation in Escherichia coli without auxotrophy

ABSTRACT: Abstract CRISPR interference (CRISPRi) was applied to enable the aerobic production of pyruvate in Escherichia coli MG1655 under glucose excess conditions by targeting the promoter regions of aceE or pdhR. Knockdown strains were cultivated in aerobic shaking flasks and the influence of inducer concentration and different sgRNA binding sites on the production of pyruvate was measured. Targeting the promoter regions of aceE or pdhR triggered pyruvate production during the exponential phase and reduced expression of aceE. In lab‐scale bioreactor fermentations, an aceE silenced strain successfully produced pyruvate under fully aerobic conditions during the exponential phase, but loss of productivity occurred during a subsequent nitrogen‐limited phase. Targeting the promoter region of pdhR enab

SUBMITTER: Ziegler M 

PROVIDER: S-EPMC8811725 | biostudies-literature | 2021 Nov

REPOSITORIES: biostudies-literature