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Literature summary extracted from

  • Mueller, M.; Kratzer, R.; Schiller, M.; Slavica, A.; Rechberger, G.; Kollroser, M.; Nidetzky, B.
    The role of Cys108 in Trigonopsis variabilis D-amino acid oxidase examined through chemical oxidation studies and point mutations C108S and C108D (2010), Biochim. Biophys. Acta, 1804, 1483-1491.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.4.3.3 expression of Strep-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3), induction by DL-Ala Trigonopsis variabilis

Protein Variants

EC Number Protein Variants Comment Organism
1.4.3.3 C108D site-directed mutagenesis, in contrast to the wild-type enzyme, the mutant releases the cofactor in a quasi-irreversible manner and is therefore not stabilized by external FAD against loss of activity. Conformational properties and kinetics of C108D, overview Trigonopsis variabilis
1.4.3.3 C108S site-directed mutagenesis, in contrast to the wild-type enzyme, the mutant releases the cofactor in a quasi-irreversible manner and is therefore not stabilized by external FAD against loss of activity. Conformational properties and kinetics of C108S, overview Trigonopsis variabilis
1.4.3.3 R110A site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme, but shows increased temperature denaturation. Release of FAD is the dominant path of thermal denaturation of R110A Trigonopsis variabilis

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.4.3.3 additional information oxidative modification of Trigonopsis variabilis D-amino acid oxidase in vivo is traceable as the conversion of Cys108 into a stable cysteine sulfinic acid, causes substantial loss of activity and thermostability of the enzyme Trigonopsis variabilis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.4.3.3 D-methionine + H2O + O2 Trigonopsis variabilis
-
 4-methylsulfanyl-2-oxobutanoate + NH3 + H2O2
-
 ?
1.4.3.3 D-methionine + H2O + O2 Trigonopsis variabilis ATCC 10679
-
 4-methylsulfanyl-2-oxobutanoate + NH3 + H2O2
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.4.3.3 Trigonopsis variabilis
-
-
-
1.4.3.3 Trigonopsis variabilis ATCC 10679
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.4.3.3 recombinant Strep-tagged wild-type and mutant enzymes from Escherichia coli strain BL21 (DE3) by ultracentrifugation, affinity and anion exchange chromatography Trigonopsis variabilis

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.4.3.3 2.2
-
 recombinant Strep-tagged wild-type enzyme, substrates are D-methionine and 6-dichloroindophenolate, pH 7.5, 37°C Trigonopsis variabilis
1.4.3.3 2.6
-
 recombinant Strep-tagged mutant R101A, substrates are D-methionine and 6-dichloroindophenolate, pH 7.5, 37°C Trigonopsis variabilis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.4.3.3 D-methionine + 2,6-dichloroindophenol
-
 Trigonopsis variabilis 4-methylsulfanyl-2-oxobutanoate + reduced 2,6-dichloroindophenol
-
 ?
1.4.3.3 D-methionine + 2,6-dichloroindophenol
-
 Trigonopsis variabilis ATCC 10679 4-methylsulfanyl-2-oxobutanoate + reduced 2,6-dichloroindophenol
-
 ?
1.4.3.3 D-methionine + H2O + O2
-
 Trigonopsis variabilis 4-methylsulfanyl-2-oxobutanoate + NH3 + H2O2
-
 ?
1.4.3.3 D-methionine + H2O + O2 with 6-dichloroindophenol as electron acceptor Trigonopsis variabilis 4-methylsulfanyl-2-oxobutanoate + NH3 + H2O2
-
 ?
1.4.3.3 D-methionine + H2O + O2
-
 Trigonopsis variabilis ATCC 10679 4-methylsulfanyl-2-oxobutanoate + NH3 + H2O2
-
 ?
1.4.3.3 D-methionine + H2O + O2 with 6-dichloroindophenol as electron acceptor Trigonopsis variabilis ATCC 10679 4-methylsulfanyl-2-oxobutanoate + NH3 + H2O2
-
 ?

Subunits

EC Number Subunits Comment Organism
1.4.3.3 homodimer
-
 Trigonopsis variabilis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.4.3.3 37
-
 assay at Trigonopsis variabilis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.4.3.3 0.84
-
 D-methionine recombinant mutant C108S, pH 7.5, 37°C, with 6-dichloroindophenolate Trigonopsis variabilis
1.4.3.3 1.5
-
 D-methionine recombinant wild-type enzyme, pH 7.5, 37°C, with 6-dichloroindophenolate Trigonopsis variabilis
1.4.3.3 2.8
-
 D-methionine recombinant mutant C108D, pH 7.5, 37°C, with 6-dichloroindophenolate Trigonopsis variabilis
1.4.3.3 75
-
 D-methionine recombinant mutant C108D, pH 7.5, 37°C, with O2 Trigonopsis variabilis
1.4.3.3 110
-
 D-methionine recombinant mutant C108S, pH 7.5, 37°C, with O2 Trigonopsis variabilis
1.4.3.3 200
-
 D-methionine recombinant wild-type enzyme, pH 7.5, 37°C, with O2 Trigonopsis variabilis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.4.3.3 7.5
-
 assay at Trigonopsis variabilis

Cofactor

EC Number Cofactor Comment Organism Structure
1.4.3.3 FAD
-
 Trigonopsis variabilis

Expression

EC Number Organism Comment Expression
1.4.3.3 Trigonopsis variabilis DL-Ala induces the enzyme up

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.4.3.3 153
-
 D-methionine recombinant mutant C108S, pH 7.5, 37°C Trigonopsis variabilis
1.4.3.3 258
-
 D-methionine recombinant mutant C108D, pH 7.5, 37°C Trigonopsis variabilis
1.4.3.3 274
-
 D-methionine recombinant wild-type enzyme, pH 7.5, 37°C Trigonopsis variabilis