Project description:In this study, we identified the O-glycosylation sites in deletion constructs of recombinant Clostridium botulinum flagellin (CbFla) expressed from the E. coli strain, EV136 (K1:K12 strains engineered by Prof. Eric Vimr’s group to accumulate CMP-sialic acid in the cytosol). The recombinant flagellins were expressed with or without co-expression of motility associated factor (Maf) (flagellin nonulosonic acid glycosyltransferase) in these strains, purified and subjected to MS/MS analysis at Taplin Biological Mass Spectrometry facility. We also studied the O-glycosylation of recombinant Clostridium botulinum flagellin (CbFla) co-expressed with Geobacillus kaustophilus Maf. We also studied the O-glycosylation of flagellin chimeras made by grafting mini-flagellin sequences on to an unrelated protein with an F-type lectin domain from Streptosporangium roseum SrNaFLD.
