Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.13.11.49 extracted from

  • Kostan, J.; Sjoeblom, B.; Maixner, F.; Mlynek, G.; Furtmueller, P.G.; Obinger, C.; Wagner, M.; Daims, H.; Djinovic-Carugo, K.
    Structural and functional characterisation of the chlorite dismutase from the nitrite-oxidizing bacterium Candidatus Nitrospira defluvii: identification of a catalytically important amino acid residue (2010), J. Struct. Biol., 172, 331-342.
    View publication on PubMed

Application

Application Comment Organism
environmental protection bacteria with Cld play significant roles in the bioremediation of industrially contaminated sites and also in wastewater treatment Nitrospira defluvii

Cloned(Commentary)

Cloned (Comment) Organism
phylogenetic analysis, expression of N-terminally His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Nitrospira defluvii

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant His-tagged wild-type and mutant enzymes with bound cyanide, 22°C, sitting drop vapour diffusion method, X-ray diffraction structure determination and analysis at 1.85-2.70 A resolution Nitrospira defluvii

Protein Variants

Protein Variants Comment Organism
R173A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme, structure determination and comparison to the wild-type enzyme Nitrospira defluvii
R173K site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme, structure determination and comparison to the wild-type enzyme Nitrospira defluvii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information steady-state, pre-steady-state, and transient-state kinetics, overview Nitrospira defluvii
0.057
-
Chlorite pH 7.0, 30°C, recombinant wild-type enzyme Nitrospira defluvii
0.107
-
Chlorite pH 7.0, 30°C, recombinant mutant R137A Nitrospira defluvii
0.818
-
Chlorite pH 7.0, 30°C, recombinant mutant R137K Nitrospira defluvii

Metals/Ions

Metals/Ions Comment Organism Structure
CN- binding kinetics of wild-type and mutant enzymes, overview Nitrospira defluvii
Fe2+ heme protein Nitrospira defluvii

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
chloride + O2 Nitrospira defluvii chlorite dismutase is a unique heme enzyme which transforms chlorite to chloride and molecular oxygen chlorite
-
r

Organism

Organism UniProt Comment Textmining
Nitrospira defluvii B3U4H7
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant N-terminally His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration Nitrospira defluvii

Reaction

Reaction Comment Organism Reaction ID
chloride + O2 = chlorite reaction mechanism and active site structure, Arg173 plays a key role in (i) controlling of ligand and substrate access and binding and (ii) in chlorite dismutation reaction, overview Nitrospira defluvii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
chloride + O2 chlorite dismutase is a unique heme enzyme which transforms chlorite to chloride and molecular oxygen Nitrospira defluvii chlorite
-
r
chloride + O2 Arg173 plays a key role in (i) controlling of ligand and substrate access and binding and (ii) in chlorite dismutation reaction, mechanism, overview. The flexible residue modulates the electrostatic potential and size of the active site entrance and might be involved in keeping transiently formed hypochlorite in place for final molecular oxygen and chloride formation Nitrospira defluvii chlorite
-
r

Subunits

Subunits Comment Organism
homopentamer each monomer of NdCld is characterized by two topologically equivalent four-stranded antiparallel beta-sheets forming a beta-barrel, flanked on both sides by six alpha-helices, overview Nitrospira defluvii

Synonyms

Synonyms Comment Organism
chlorite dismutase
-
Nitrospira defluvii
CLD
-
Nitrospira defluvii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25 30 assay at Nitrospira defluvii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
7.1
-
Chlorite pH 7.0, 30°C, recombinant mutant R137A Nitrospira defluvii
21
-
Chlorite pH 7.0, 30°C, recombinant mutant R137K Nitrospira defluvii
35
-
Chlorite pH 7.0, 30°C, recombinant wild-type enzyme Nitrospira defluvii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
assay at Nitrospira defluvii

Cofactor

Cofactor Comment Organism Structure
heme
-
Nitrospira defluvii

General Information

General Information Comment Organism
evolution active site conservation and evolution of Cld, overview Nitrospira defluvii
additional information Candidatus Nitrospira defluvii is a key nitrifier in biological wastewater treatment, molecular mechanism of chlorite detoxification, overview. The residue corresponding to Arg173 is conserved in all known active forms of Cld and propose it as a marker for Cld activity in yet uncharacterized Cld-like proteins Nitrospira defluvii

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
25000
-
Chlorite pH 7.0, 30°C, recombinant mutant R137K Nitrospira defluvii
66000
-
Chlorite pH 7.0, 30°C, recombinant mutant R137A Nitrospira defluvii
600000
-
Chlorite pH 7.0, 30°C, recombinant wild-type enzyme Nitrospira defluvii