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Urease from a potentially pathogenic coccoid isolate: purification, characterization, and comparison to other

S G Lee1, D H Calhoun

  • 1The Graduate School and University Center and Department of Chemistry, City College of New York, The City University of New York, New York 10031, USA.

Infection and Immunity
|October 8, 1997
PubMed
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This study purified urease from a gastric pathogen, finding it

Area of Science:

  • Microbiology
  • Enzymology
  • Pathogenesis

Background:

  • Strain SL100 is a gram-positive coccoid isolate from gastric biopsies, possessing a mucin adhesin.
  • This organism is a potential pathogen associated with gastric disorders.
  • Urease is a significant protein component of SL100, associated with its cell wall.

Purpose of the Study:

  • To purify and characterize the urease enzyme from strain SL100.
  • To investigate the stability and kinetic properties of the purified urease.
  • To compare the SL100 urease with those from other microbial species, particularly Helicobacter.

Main Methods:

  • Purification of urease using a strategy involving nickel sulfate stabilization.
  • Gel electrophoresis to assess purity and subunit composition.

Related Experiment Videos

  • Enzyme kinetics assays (K(m), V(max)) on whole cells and purified enzyme.
  • Acid stability assays.

    • Main Results:

    • Urease was purified 138-fold to apparent homogeneity with a specific activity of 1,120 U/mg.
    • Nickel was essential for urease stability during purification and storage.
    • The native urease is a 260 kDa enzyme composed of 65 kDa and 21 kDa subunits.
    • The enzyme demonstrated remarkable acid stability (retaining activity at pH 1.3).
    • Kinetic parameters and subunit composition resemble Helicobacter ureases.

    Conclusions:

    • The urease from strain SL100 is a stable, acid-tolerant enzyme with characteristics similar to Helicobacter ureases.
    • Nickel is crucial for maintaining urease activity and stability.
    • The urease likely plays a role in the organism's adaptation and colonization of the stomach, potentially acting as a virulence factor.