Assaying <i>Paenibacillus alvei</i> CsaB-Catalysed Ketalpyruvyltransfer to Saccharides by Measurement of Phosphate Release.

ABSTRACT: Ketalpyruvyltransferases belong to a widespread but little investigated class of enzymes, which utilise phosphoenolpyruvate (PEP) for the pyruvylation of saccharides. Pyruvylated saccharides play pivotal biological roles, ranging from protein binding to virulence. Limiting factors for the characterisation of ketalpyruvyltransferases are the availability of cognate acceptor substrates and a straightforward enzyme assay. We report on a fast ketalpyruvyltransferase assay based on the colorimetric detection of phosphate released during pyruvyltransfer from PEP onto the acceptor via complexation with Malachite Green and molybdate. To optimise the assay for the model 4,6-ketalpyruvyl::ManNAc-transferase CsaB from Paenibacillus alvei, a β-d-ManNAc-α-d-GlcNAc-diphosphoryl-11-phenoxyundecyl

SUBMITTER: Hager-Mair FF 

PROVIDER: S-EPMC8615578 | biostudies-literature | 2021 Nov

REPOSITORIES: biostudies-literature