Flavobacterium heparinum sulphamidase for D-glucosamine sulphamate. Purification and characterisation.
A novel assay has been developed for 2-deoxy-2-sulphamido-D-glucose (GlcNS) sulphamidase from Flavobacterium heparinum. This has enabled the 1930-fold purification of the enzyme from a soluble fraction of bacterial homogenate. From SDS/polyacrylamide gel electrophoresis the enzyme was shown to have a relative molecular mass of 81,500. Ca2+ was essential for enzyme activity. Inorganic phosphate and sulphate inhibited activity by 28% and 29% respectively at 5 mmol dm-3. The purified sulphamidase had a pH optimum of 7.0 and a Km of 8.32 mumol dm-3 for GlcNS. The degradation of 2-deoxy-2-sulphamido-6-O-sulpho-D-glucose (GlcNS-6S) was also re-investigated. The two sulphate groups were hydrolysed sequentially in a single non-bifurcate manner, in contrast to previous reports [Dietrich, C.P., Silva, M.E. and Michelacci, Y.M. (1973) J. Biol. Chem. 248, 6408-6415].
D000838:Anions / Q000494:pharmacology; D002412:Cations / Q000494:pharmacology; D005417:Flavobacterium / Q000201:enzymology*; D006867:Hydrolases / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D006868:Hydrolysis; D009682:Magnetic Resonance Spectroscopy; D013429:Sulfatases / Q000378:metabolism*
D000838:Anions; D002412:Cations; D006867:Hydrolases; D013429:Sulfatases; C028553:disulfoglucosamine-6-sulfatase; C022708:N-sulfoglucosamine sulfohydrolase