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Methods for the measurement of a bacterial enzyme activity in cell lysates and extracts

  • Brendan P. Burns
  • George L. Mendz
  • Stuart L. Hazell
Open Access
Article

Abstract

The kinetic characteristics and regulation of aspartate carbamoyltransferase activity were studied in lysates and cell extracts of Helicobacter pylori by three different methods. Nuclear magnetic resonance spectroscopy, radioactive tracer analysis, and spectrophotometry were employed in conjunction to identify the properties of the enzyme activity and to validate the results obtained with each assay. NMR spectroscopy was the most direct method to provide proof of ACTase activity; radioactive tracer analysis was the most sensitive technique and a microtitre-based colorimetric assay was the most cost-and time-efficient for large scale analyses. Freeze-thawing was adopted as the preferred method for cell lysis in studying enzyme activity in situ. This study showed the benefits of employing several different complementary methods to investigate bacterial enzyme activity.

Keywords

Nuclear Magnetic Resonance Antipyrine Nuclear Magnetic Resonance Spectroscopy Biological Procedure Carbamoyl Phosphate 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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Copyright information

© Springer 1998

Authors and Affiliations

  • Brendan P. Burns
    • 1
  • George L. Mendz
    • 2
  • Stuart L. Hazell
    • 1
  1. 1.School of Microbiology and ImmunologyThe University of New South WalesSydneyAustralia
  2. 2.School of Biochemistry and Molecular GeneticsThe University of New South WalesSydneyAustralia

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