Methods for the Pasteurella Glycosaminoglycan Synthases: Enzymes that Polymerize Hyaluronan, Chondroitin, or Heparosan Chains

Abstract

Multiple glycosyltransferases (GTases) that produce glycosaminoglycans (GAGs) have been identified; ­several distinct putative architectures and catalytic abilities have been noted from microbes and vertebrates. Here the preparation and use of a class of enzymes (Class II) from the gram-negative bacterium, Pasteurella multocida, with utility in chemoenzymatic synthesis of GAGs is described. The analyses of sugar products include thin layer chromatography (TLC), matrix-assisted laser desorption ionization mass spectroscopy (MALDI-ToF MS), and polyacrylamide gel electrophoresis (PAGE). The related Chapter 18 is focused on larger molecular weight GAGs analyses as well as the Class I membrane streptococcal and mammalian HA synthases.