Purification and properties of urease from Sporobolomyces roseus.
Urease (EC 3.5.1.5) catalyses the hydrolysis of urea to ammonia and carbon dioxide. The enzyme from Sporobolomyces roseus was enriched 780-fold and purified to apparent homogeneity using heat treatment, ion exchange chromatography on Q-Sepharose fast flow, hydrophobic interaction chromatography on Phenyl-Sepharose, size exclusion chromatography on Sephacryl S 300 HR, and ion exchange chromatography on MonoQ. Analysis of the purified enzyme by SDS-PAGE demonstrated the presence of subunits with a molecular weight of 90 (+/- 4) kDa. The M(r) of the native enzyme was estimated by size exclusion chromatography to be 340 (+/- 30) kDa, suggesting a tetrameric structure different from other ureases isolated so far from both prokaryotes and eukaryotes. The enzyme was heat-stable, showing no loss of activity after incubation at 70 degrees C for 15 min. The highest urease activities were observed after growth on media containing urea as the sole source of nitrogen.
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Fachrichtung 13.3 Mikrobiologie, Universität des Saarlandes, Saarbrücken, Germany.
D002614:Chelating Agents / Q000494:pharmacology; D002852:Chromatography, Ion Exchange; D003470:Culture Media; D004492:Edetic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000302:isolation & purification* / Q000378:metabolism; D006358:Hot Temperature; D006877:Hydroxamic Acids / Q000494:pharmacology; D007700:Kinetics; D009532:Nickel; D009584:Nitrogen / Q000378:metabolism; D014508:Urea / Q000378:metabolism; D014510:Urease / Q000302:isolation & purification* / Q000378:metabolism; D015003:Yeasts / Q000201:enzymology*
D002614:Chelating Agents; D003470:Culture Media; D005656:Fungal Proteins; D006877:Hydroxamic Acids; C006358:acetohydroxamic acid; D009532:Nickel; D014508:Urea; D004492:Edetic Acid; D014510:Urease; D009584:Nitrogen