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Archives of Microbiology

, Volume 166, Issue 5, pp 321–326 | Cite as

Purification and characterization of chlorite dismutase: a novel oxygen-generating enzyme

  • C. G. van Ginkel
  • G. B. Rikken
  • A. G. M. Kroon
  • S. W. M. Kengen
Original paper

Abstract

A novel enzyme that catalyzes the disproportionation of chlorite into chloride and oxygen was purified from a gram-negative bacterium, strain GR-1 to homogeneity. A four-step purification procedure comprising Q-Sepharose, hydroxyapatite, and phenyl-Superose chromatography and ultrafiltration resulted in a 13.7-fold purified enzyme with a final specific activity of 2.0 mmol min–1 (mg protein)–1. The dismutase obeyed Michaelis-Menten kinetics. The V max and K m calculated for chlorite were 2,200 U (mg protein)–1 and 170 μM, respectively. Dismutase activity was inhibited by hydroxylamine, cyanide, and azide, but not by 3-amino-1,2,4-triazole. Chlorite dismutase had a molecular mass of 140 kDa and consisted of four 32-kDa subunits. The enzyme was red-colored and had a Soret peak at 392 nm. Per subunit, it contained 0.9 molecule of protoheme IX and 0.7 molecule of iron. Chlorite dismutase displayed maxima for activity at pH 6.0 and 30° C.

Key words Chlorite Oxygen Chloride Chlorite dismutase Homotetramer Heme iron Disproportionation 

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Copyright information

© Springer-Verlag Berlin Heidelberg 1996

Authors and Affiliations

  • C. G. van Ginkel
    • 1
  • G. B. Rikken
    • 1
  • A. G. M. Kroon
    • 1
  • S. W. M. Kengen
    • 2
  1. 1.Akzo Nobel Central Research, Analytical and Environmental Chemistry Department, Velperweg 76, 6800 SB Arnhem, The Netherlands Tel. +31-26-366-2634; Fax +31-26-366-2528 e-mail: Kees.C.G.vanginkel@Akzo.NLNL
  2. 2.Wageningen Agricultural University, Department of Microbiology, Hesselink van Suchtelenweg 4, 6703 CT Wageningen, The NetherlandsNL

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