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bioRxiv · 10.1101/2025.07.17.665469

Deletion of major shell protein of ethanolamine utilization microcompartment reduces intrinsic antibiotic resistance, biofilm and intracellular survival of Salmonella Typhimurium

Bhalerao, M. R.·Davkhar, A. S.·Sawant, A. R.·Ghosh, A. S.·Harris, T. N.·Bobik, T.·Saha, A.·Dastager, S.·Agawane, S. A.·Chowdhury, C.
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Abstract

With the high rise in Salmonella infection and emergence of antibiotic-resistant variants, developing a novel strategy to control the pathogen is imperative. Earlier studies revealed that Salmonella deploys ethanolamine (EA) metabolic machinery to disseminate in the intestine. Salmonella with a defect in EA metabolism manifests with lower intestinal colonization efficiency. Remarkably, the potential of EA metabolism as a therapeutic target is yet to explore. Our study revealed that supplementation of EA and vitamin B12 in both rich and minimal media enhanced biofilm formation, increased motility, and increased tolerance of Salmonella to some antibiotics. Conversely, mutants deficient in EA metabolic enzymes exhibited no physiological fitness. In Salmonella, EA metabolic enzymes are localized within a proteinaceous microcompartment (MCP) shell composed of thousands of copies of shell proteins encoded by five genes from the eut operon. Fascinatingly, bacterial cells with defective MCP shell due to mutation in the major shell proteins showed enhanced susceptibility towards a number of antibiotics in minimal media. The mutants were unable to form biofilm, produced lower curli expression and were defective in flagellar motility. Also, mutation in one of the major shell proteins reduced intramacrophagic viability of Salmonella. Notably, phenotypes were restored upon ectopic expression of corresponding genes. It was evident that mutation in the MCP shell proteins downregulated the expression of genes related to pathogenicity. Overall, this study sheds new light on understanding the relationship between EA metabolism and bacterial physiology that would pave the way for developing novel therapeutic interventions against Salmonella.

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Bhalerao, M. R., Davkhar, A. S., Sawant, A. R., Ghosh, A. S., Harris, T. N., Bobik, T., Saha, A., Dastager, S., Agawane, S. A., Chowdhury, C.. 2025-07-18. Deletion of major shell protein of ethanolamine utilization microcompartment reduces intrinsic antibiotic resistance, biofilm and intracellular survival of Salmonella Typhimurium. https://doi.org/10.1101/2025.07.17.665469

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