Isolation of a high specific activity pink, monomeric nitrous oxide reductase from Achromobacter cycloclastes.
Nitrous oxide reductase from Achromobacter cycloclastes has been purified to homogeneity under aerobic conditions via DEAE-cellulose, phenyl-Sepharose, hydroxyapatite, and Sephacryl S-200 chromatography. It consists of a single polypeptide of MW 72 kDa, and contains 3.8 +/- 0.1 copper atoms per molecule. The enzyme is pink as isolated, yet exhibits a specific activity (86 U/mg) that is ca. 40 times greater than that observed for other N2O reductases under similar conditions. Double integration of the anomalous EPR spectrum at 77K showed the presence of 2.0 +/- 0.1 spins per molecule, implying the presence of EPR-silent copper atoms and/or spin-coupled mixed-valent centers.
Hulse
C L
CL
Department of Chemistry, University of Virginia, Charlottesville 22901.
Averill
B A
BA
D000421:Alcaligenes / Q000201:enzymology*; D000596:Amino Acids / Q000032:analysis; D000693:Anaerobiosis; D001426:Bacterial Proteins / Q000302:isolation & purification; D004578:Electron Spin Resonance Spectroscopy; D046911:Macromolecular Substances; D008667:Metalloproteins / Q000302:isolation & purification; D010088:Oxidoreductases / Q000302:isolation & purification*; D013057:Spectrum Analysis
D000596:Amino Acids; D001426:Bacterial Proteins; D046911:Macromolecular Substances; D008667:Metalloproteins; D010088:Oxidoreductases; C022763:nitrous oxide reductase