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

Biological Procedures Online volume 1, pages 17–26 (1998)Cite this 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.

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Authors and Affiliations

  1. School of Microbiology and Immunology, The University of New South Wales, 2052, Sydney, Australia

    Brendan P. Burns & Stuart L. Hazell

  2. School of Biochemistry and Molecular Genetics, The University of New South Wales, 2052, Sydney, Australia

    George L. Mendz

Authors
  1. Brendan P. Burns
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  2. George L. Mendz
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  3. Stuart L. Hazell
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Correspondence to Brendan P. Burns.

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Burns, B.P., Mendz, G.L. & Hazell, S.L. Methods for the measurement of a bacterial enzyme activity in cell lysates and extracts. Biol Proced Online 1, 17–26 (1998). https://doi.org/10.1251/bpo5

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  • DOI: https://doi.org/10.1251/bpo5

Keywords

  • Nuclear Magnetic Resonance
  • Antipyrine
  • Nuclear Magnetic Resonance Spectroscopy
  • Biological Procedure
  • Carbamoyl Phosphate

Biological Procedures Online

ISSN: 1480-9222

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