Folia Microbiologica

, Volume 42, Issue 5, pp 431–440 | Cite as

Identification and characterization of a heat-labile type I glutamine synthetase fromStreptomyces cinnamonensis

  • K. T. Nguyen
  • L. T. Nguyen
  • O. Benada
  • V. Běhal
Papers
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Abstract

Streptomycetes have two distinct glutamine synthetases (GS): a heat-stable dodecameric GSI and a heat-labile octameric GSII. A heat-inactivated GS activity was detected in crude extracts ofStreptomyces cinnamonensis cells grown with nitrate or glutamate as the nitrogen source. The purified enzyme obtained from crude extracts of the nitrate-grown cells after affinity and anion-exchange chromatography was also heat-labile; it was inactivated by 80 % when incubated at 50 °C for 1 h. However, the enzyme has properties typical of GSI and similar with those of the heat-stable GSI purified fromS. aureofaciens: It is composed of twelve subunits, each ofM 55 kDa, and has a native molar mass of 625 kDa and an isoelectric point at pH 4.2. In addition, its activity is regulated by reversible adenylylation. Mg2+ and NaCl but not Mn2+ protected the purified enzyme from thermal inactivation, and both NaCl and Mn2+ or Mg2+ stabilized its activity at 4–8 °C. As compared with GSI fromS. aureofaciens, theS. cinnamonensis enzyme was cleaved more extensively during SDS-PAGE, was less sensitive to feedback inhibitors, and similarly affected by divalent cations. TheK m values were 12.5 mmol/L forl-glutamate, 0.1 for NH 4 + , 1.25 for ATP, 18.5 forl-glutamine, 3.3 for hydroxylamine and 0.087 for ADP. To our best knowledge, this is the first report of a heatlabile GSI from any source.

Keywords

Streptomyces Crude Extract Glutamine Synthetase Carbamyl Phosphate Glutamine Synthetase Protein 
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

© Institute of Microbiology, Academy of Sciences of the Czech Republic 1997

Authors and Affiliations

  • K. T. Nguyen
    • 1
  • L. T. Nguyen
    • 1
  • O. Benada
    • 1
  • V. Běhal
    • 1
  1. 1.Institute of MicrobiologyAcademy of Sciences of the Czech RepublicPrague 4Czech Republic

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