Purification and properties of urease from Sporobolomyces roseus

Antonie Van Leeuwenhoek. 1995 Oct;68(3):209-14. doi: 10.1007/BF00871817.

Abstract

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.

MeSH terms

  • Chelating Agents / pharmacology
  • Chromatography, Ion Exchange
  • Culture Media
  • Edetic Acid / pharmacology
  • Electrophoresis, Polyacrylamide Gel
  • Fungal Proteins / isolation & purification*
  • Fungal Proteins / metabolism
  • Hot Temperature
  • Hydroxamic Acids / pharmacology
  • Kinetics
  • Nickel
  • Nitrogen / metabolism
  • Urea / metabolism
  • Urease / isolation & purification*
  • Urease / metabolism
  • Yeasts / enzymology*

Substances

  • Chelating Agents
  • Culture Media
  • Fungal Proteins
  • Hydroxamic Acids
  • acetohydroxamic acid
  • Nickel
  • Urea
  • Edetic Acid
  • Urease
  • Nitrogen