CRISPR interference of nucleotide biosynthesis improves production of a single-domain antibody in Escherichia coli.

ABSTRACT: Growth decoupling can be used to optimize the production of biochemicals and proteins in cell factories. Inhibition of excess biomass formation allows for carbon to be utilized efficiently for product formation instead of growth, resulting in increased product yields and titers. Here, we used CRISPR interference to increase the production of a single-domain antibody (sdAb) by inhibiting growth during production. First, we screened 21 sgRNA targets in the purine and pyrimidine biosynthesis pathways and found that the repression of 11 pathway genes led to the increased green fluorescent protein production and decreased growth. The sgRNA targets pyrF, pyrG, and cmk were selected and further used to improve the production of two versions of an expression-optimized sdAb. Proteomics analysis of

SUBMITTER: Landberg J 

PROVIDER: S-EPMC7818426 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature