title: Purification of surface-associated urease from Helicobacter pylori.
abstract: Helicobacter pylori colonizes the human gastric mucosa and produces large amounts of urease. The enzyme was extracted from the bacteria by distilled water and purified by gel-permeation (Sephacryl S-300), anion-exchange chromatography (Mono Q) and a second gel-permeation (Superdex 200). Urease enzyme activity was detected with a spectrophotometic assay based on phenol red. The optimal pH for anion-exchange was 6.9. The recovery of urease was 55-75%, purity 93-98% and the overall protein recovery 0.8-1.4%. The urease in the final extract still had enzymatic activity and showed the typical subunits of Mr 66000 and Mr 30000 when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
authors.affiliation: Department of Clinical Microbiology, Hygiene-Institute, University of Vienna, Austria. elmar.rokita@akh-wien.ac.at
authors.forename: E
authors.initials: E
authors.lastname: Rokita
authors.forename: A
authors.initials: A
authors.lastname: Makristathis
authors.forename: A M
authors.initials: AM
authors.lastname: Hirschl
authors.forename: M L
authors.initials: ML
authors.lastname: Rotter
mesh_terms: D015153:Blotting, Western; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D016480:Helicobacter pylori / Q000201:enzymology*; D014510:Urease / Q000302:isolation & purification*
chemical_list: D014510:Urease