PT - JOURNAL ARTICLE AU - Zhe Zeng AU - Lucas M. Wijnands AU - Sjef Boeren AU - Eddy J. Smid AU - Richard A. Notebaart AU - Tjakko Abee TI - Impact of vitamin B12 on rhamnose metabolism, stress defense and in-vitro virulence of <em>Listeria monocytogenes</em> AID - 10.1101/2021.08.26.457850 DP - 2021 Jan 01 TA - bioRxiv PG - 2021.08.26.457850 4099 - http://biorxiv.org/content/early/2021/08/26/2021.08.26.457850.short 4100 - http://biorxiv.org/content/early/2021/08/26/2021.08.26.457850.full AB - Listeria monocytogenes is a facultative anaerobe which can cause a severe food-borne infection known as listeriosis. Rhamnose is a deoxyhexose sugar abundant in a range of environments, including the human intestine, and can be degraded by L. monocytogenes in aerobic and anaerobic conditions into lactate, acetate and 1,2-propanediol. Our previous study showed that addition of vitamin B12 stimulates anaerobic growth of L. monocytogenes on rhamnose due to the activation of bacterial microcompartment (BMC)-dependent 1,2-propanediol utilization with concomitant production of propionate and propanol. Notably, anaerobic propanediol metabolism has been linked to virulence of enteric pathogens including Salmonella spp. and L. monocytogenes. In this study we investigate the impact of B12 on aerobic and anerobic growth of L. monocytogenes on rhamnose, and observed growth stimulation and pdu BMC activation only in anaerobically grown cells with B12 added to the medium. Comparative Caco-2 virulence assays, showed that these pdu BMC induced cells have significantly higher translocation efficiency compared to aerobically grown cells (without and with added B12) and non-induced anaerobically grown cells, while adhesion and invasion capacity is similar for all cells. Comparative proteomics analysis showed specific and overlapping responses linked to metabolic shifts, activation of stress defense proteins and virulence factors, with RNA polymerase sigma factor SigL; teichoic acids export ATP-binding protein, TagH; DNA repair and protection proteins RadA and DPS; and glutathione synthase GshAB previously linked to activation of virulence response in L. monocytogenes, uniquely upregulated in anaerobically rhamnose grown pdu BMC induced cells. Our results shed new light into B12 impact on L. monocytogenes competitive fitness and virulence.Competing Interest StatementThe authors have declared no competing interest.