Harnessing the acceptor substrate promiscuity of Clostridium botulinum Maf glycosyltransferase to glyco-engineer mini-flagellin protein chimeras

ABSTRACT: 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.

INSTRUMENT(S):

ORGANISM(S): Clostridium Botulinum Str. Langelan F

SUBMITTER: Ramya Chakravarthy  

LAB HEAD: Ramya Chakravarthy

PROVIDER: PXD044191 | Pride | 2024-08-22

REPOSITORIES: Pride