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1.13.11.49: chlorite O2-lyase

This is an abbreviated version!
For detailed information about chlorite O2-lyase, go to the full flat file.

Word Map on EC 1.13.11.49

Reaction

chloride
+
O2
=
Chlorite

Synonyms

chlorite dismutase, CLD, Cyan7425_1434, dimutase, chlorite, HemQ, Pfam chlorite dismutase, PitA

ECTree

     1 Oxidoreductases
         1.13 Acting on single donors with incorporation of molecular oxygen (oxygenases)
             1.13.11 With incorporation of two atoms of oxygen
                1.13.11.49 chlorite O2-lyase

Purification

Purification on EC 1.13.11.49 - chlorite O2-lyase

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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
anion exchange column chromatography and S-200 HR gel filtration
cell pellet suspended in 0.05 M Tris-HCl buffer, pH 9.5 and 0.05 EDTA for 30 min at room temperature, centrifuged, supernatant ultracentrifuged, red supernatant containing periplasmic fraction separated from pellet, diluted with 10 mM Tris-HCl buffer, pH 7.2, purification with ÄKTAfplc, diluted supernatant loaded onto hydroxyapatite column with 10 mM Tris-HCl, pH 7.2, enzyme elutes at the start of a linear gradient of 10 mM Tris-Hcl, pH 7.2 to 450 mM potassium phosphate, pH 7.2, storage at -20°C
Stutzerimonas chloritidismutans
-
cell-free extracts are prepared from strain AW-1 cells grown in anaerobic medium with n-decane as sole carbon and energy source and chlorate as electron acceptor, centrifugation, storage of extracts under N2 gas at 4°C
Stutzerimonas chloritidismutans
-
centrifuged cell extract, cells grown in AW-1-sulfate medium with benzene and chlorate or benzene and oxygen
-
Co2+ chelating resin column chromatography
-
HiLoad 16/60 Superdex 200 prep-grade column, equilibrated with 20 mM Tris-HCl (pH 7.5) and 135 mM NaCl to determin molecular Stokes radius
His-trap affinity column chromatography and Superdex 200 gel filtration
HisTrap column chromatography
-
Ni Sepharose 6 column chromatography
Ni-NTA Sepharose column chromatography
-
Ni-Sepharose 6 column chromatography
-
recombinant enzyme from Escherichia coli by anion exchange chromatography and gel filtration
-
recombinant His-tagged enzyme from Escherichia coli strain Tuner (DE3) by affinity chromatography
recombinant His-tagged enzyme from Escherichia coli strain Tuner (DE3) by nickel affinity chromatography and gel filtration
recombinant mature Cld from Tuner(DE3) Escherichia coli cells by anion exchange chromatography and gel filtration
-
recombinant N-terminally His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration
recombinant Strep-tagged enzyme from Escherichia coli strain Tuner (DE3) by affinity chromatography, the Strep-II-tag is fully cleaved off using TEV-protease, followed by gel filtration
-
the EPR spectroscopic signature of the as-purified Cld samples is affected by the buffer composition. Potassium phosphate buffer is the only buffer that affects neither the spectral nor the kinetic properties of Cld