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Summary: Glutamine synthetase (GS) (EC 6.3.1.2) was purified from an Escherichia coli glnA deletion strain containing the Thiobacillus ferrooxidans GS structural gene. The apparent Mr of the cloned T. ferrooxidans GS subunit was approximately 60000. This indicates a particle Mr for the undissociated enzyme of 720000, assuming the enzyme is the typical dodecamer. Electron microscopy of purified GS revealed characteristic disc shaped molecules with central holes. The cloned T. ferrooxidans GS was regulatedby Mg2+ or Mn2+,adenylylation, and nitrogen source but was not affected by feedback modifiers. The GS has a γ-glutamyltransferase isoactivity point of pH 7.71.
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