Dr. Thomas Plummer has found that gram-negative bacterium secretes into culture medium two proteases, an amidase called PNGase 2 F, and endoglycosidases, Endo F2 and F3. The genes for the glycosidases have been cloned and their nucleotide sequences determined. Native Endo F2 and F3 run larger on SDS-PAGE than the enzymes expressed in E. Coli indicating glycosylation of the native forms. An O-linked glycopeptide has been isolated from Endo F2 and its structure characterized by Dr. Reinhold. A comprehensive analysis of these enzymes will include a determination of their substrate specificity and their glycan structures using mass spectrometry. An effort will be made to determine whether the unusual chain-cleaving enzymes are produced by other strains of Flavobacterium or other prokaryotes.
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