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Klebsiella pneumoniae pks gene cluster

ABSTRACT: Prevalence and characteristics of pks gene cluster harboring Klebsiella pneumoniae from bloodstream infection in China

PROVIDER: PRJEB32094 | ENA |

REPOSITORIES: ENA

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Pseudomonas species. PKS

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Project description:Polyketide synthases (PKSs) are modular enzymes with exceptional potential as biocatalysts for producing non-native compounds. Here, we report the first PKS-based pathway for adipic acid (AA), an industrial monomer for nylon production, by engineering one of the most extensively hybridized PKS systems to date. Using a retrobiosynthetic approach, we identified EtnB, a succinyl-CoA loading module that uniquely retains the terminal carboxyl group, enabling access to dicarboxylic polyketide products, rarely produced by canonical PKSs. EtnB was coupled to a fully reducing extension module through an engineered communication linker, which improved ACP–KS interactions, enhanced titers, and demonstrated selective S-CoA loading in vivo. This construct integrates genes from five organisms, seven PKS modules joined across six non-natural junctions, and functions in both Escherichia coli and Pseudomonas putida. Additional engineering that included AT domain swaps, optimization of extender unit supply, and host strain metabolic rewiring further improved AA titers into the low mg/L range. Together, this work demonstrates that highly chimeric PKSs can be rendered functional through rational design, expands the PKS toolkit with a carboxyl-retaining loading module, and establishes a versatile platform for engineering diacids and other non-canonical products through PKS pathways.

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Project description:Colorectal cancer (CRC)–associated pathobionts such as Enterotoxigenic Bacteroides fragilis (ETBF) and colibactin-producing (pks⁺) Escherichia coli disrupt epithelial barrier function, promote chronic inflammation, and induce genotoxic stress. To investigate how these pathogenic interactions with the host epithelium can be modulated, we employed patient-derived normal human colon organoids as an ex vivo model. Two lytic bacteriophages, Bac-FRP-3 (targeting ETBF) and Esc-COP-23 (targeting pks⁺ E. coli), were applied to bacterial cultures, and the resulting phage-conditioned supernatants were used to challenge organoids. Exposure to ETBF and pks⁺ E. coli induced strong pro-inflammatory transcriptional responses, DNA damage signaling, and epithelial disruption, whereas phage treatment attenuated these effects. Transcriptomic profiling revealed restoration of inflammatory and DNA damage–related gene expression, while confocal imaging confirmed recovery of epithelial junctional integrity and reduced DNA double-strand breaks. These findings highlight phage-mediated modulation of host–pathobiont interactions and provide mechanistic insight into how targeted viral predation can reshape pathogenic impacts within the human intestinal environment.

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