Tuning a high performing multiplexed-CRISPRi Pseudomonas putida strain to further enhance indigoidine production.

ABSTRACT: In this study, a 14-gene edited Pseudomonas putida KT2440 strain for heterologous indigoidine production was examined using three distinct omic datasets. Transcriptomic data indicated that CRISPR/dCpf1-interference (CRISPRi) mediated multiplex repression caused global gene expression changes, implying potential undesirable changes in metabolic flux. ¹³C-metabolic flux analysis (¹³C-MFA) revealed that the core P. putida flux network after CRISPRi repression was conserved, with moderate reduction of TCA cycle and pyruvate shunt activity along with glyoxylate shunt activation during glucose catabolism. Metabolomic results identified a change in intracellular TCA metabolites and extracellular metabolite secretion profiles (sugars and succinate overflow) in t

SUBMITTER: Czajka JJ 

PROVIDER: S-EPMC9494242 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature