CRISPR interference-guided modulation of glucose pathways to boost aconitic acid production in Escherichia coli.

ABSTRACT:

Background

One major mission of microbial breeding is high-level production of desired metabolites. Overproduction of intermediate metabolites in core pathways is challenging as it may impair cell growth and viability.

Results

Here we report that aconitic acid, an intermediate metabolite in tricarboxylic acid (TCA) cycle, can be overproduced by an engineered CRISPR interference (CRISPRi) system in Escherichia coli. This CRISPRi system was designed to simultaneously target pyruvate kinase (PK) and isocitrate dehydrogenase (IDH), two enzymes in glycolytic pathway and TCA cycle, respectively. Reverse transcription and quantitative PCR and enzyme activity assays showed that this engineered CRISPRi system significantly repressed the genes encoding IDH and PK, resulting in simulta

SUBMITTER: Li Q 

PROVIDER: S-EPMC7470443 | biostudies-literature | 2020 Sep

REPOSITORIES: biostudies-literature