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Literature summary extracted from

  • Rungsrisuriyachai, K.; Gadda, G.
    On the role of histidine 351 in the reaction of alcohol oxidation catalyzed by choline oxidase (2008), Biochemistry, 47, 6762-6769.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.3.17 expressed in Escherichia coli, wild-type and H351A variant, plasmid pET/codAmg1-H351A Arthrobacter globiformis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.3.17 active site of wild-type choline oxidase, resolution of 1.86 A Arthrobacter globiformis

Protein Variants

EC Number Protein Variants Comment Organism
1.1.3.17 H351A generated by site-directed mutagenesis Arthrobacter globiformis

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.3.17 glycine betaine pH dependence of the kcat and kcat/Km values for glycine betaine inhibition shown, inhibition of H351A enzyme maximal at low pH and reduced with increasing pH Arthrobacter globiformis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.3.17 choline + O2 Arthrobacter globiformis
-
 betaine aldehyde + H2O2
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.3.17 Arthrobacter globiformis Q7X2H8 strain ATCC 8010
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.3.17 wild-type and mutant protein, purified enzymes have 50-70% of the enzyme-bound flavin cofactor as an air-stable anionic flavosemiquinone, converts slowly to oxidized state by extensive dialysis at pH 6 and 4°C Arthrobacter globiformis

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.3.17 additional information
-
 pH dependence of substrate deuterium isotope effects and of the kcat and kcat/Km values for choline in the mutant variant H351A determined, steady-state kinetic parameters compared between H351A mutant and wild-type, His351 important but not essential for the reductive half-reaction of choline oxidation, contributes to substrate binding and to the overall polarity of the active site Arthrobacter globiformis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.3.17 choline + O2
-
 Arthrobacter globiformis betaine aldehyde + H2O2
-
 ?
1.1.3.17 choline + O2 choline shown to be a slow substrate for H351A variant, His351 residue important for substrate binding and hydride transfer reaction Arthrobacter globiformis betaine aldehyde + H2O2
-
 ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.3.17 25
-
 activity assay at Arthrobacter globiformis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.3.17 additional information
-
 additional information overall turnover of mutant variant about 60-fold decreased with choline compared to wild-type enzyme Arthrobacter globiformis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.3.17 10
-
 activity assay at, in 50 mM sodium pyrophosphate, neutral hydroquinone species of the flavin stabilized in the H351A enzyme at Arthrobacter globiformis

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.3.17 5 10 varying concentrations of both choline and oxygen, between pH 7 and pH 10, Km values for oxygen less than 15 microM Arthrobacter globiformis

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.3.17 FAD His351 stabilizes transition state for hydride transfer reaction to flavin, is not involved in the activation of the reduced flavin for reaction with oxygen Arthrobacter globiformis