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Literature summary extracted from

  • Job, V.; Molla, G.; Pilone, M.S.; Pollegioni, L.
    Overexpression of a recombinant wild-type anf His-tagged Bacillus subtilis glycine oxidase in Escherichia coli (2002), Eur. J. Biochem., 269, 1456-1463.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.4.3.19 production of a recombinant plasmid, pT7-GO by insertion of the DNA encoding for glycine oxidase into the multiple cloning site of the expression vector pT7.7. The pT7-glycine oxidase encodes a fully active fusion protein with six additional residues at the N-terminus of glycine oxidase. In BL21(DE3)pLysS Escherichia coli cells, and under optimal isopropyl thio-beta-D-galactoside induction conditions, soluble and active chimeric glycine oxidase is expressed up to 1.14 U per mg of cell. An N-terminally His-tagged glycine oxidase is also successfully expressed in Escherichia coli as a soluble protein and a fully active holoenzyme Bacillus subtilis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.4.3.19 46600
-
 4 * 46600, recombinant glycine oxidase, SDS-PAGE Bacillus subtilis
1.4.3.19 49400
-
 4 * 49400, recombinant His-tagged chimeric glycine oxidase Bacillus subtilis
1.4.3.19 166400
-
 recombinant His-tagged chimeric glycine oxidase, gel filtration Bacillus subtilis
1.4.3.19 187600
-
 recombinant glycine oxidase, gel filtration Bacillus subtilis

Organism

EC Number Organism UniProt Comment Textmining
1.4.3.19 Bacillus subtilis
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.4.3.19 wild-type enzyme and His-tagged chimeric glycine oxidase Bacillus subtilis

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.4.3.19 0.568
-
 wild-type enzyme Bacillus subtilis
1.4.3.19 1.06
-
 His-tagged chimeric glycine oxidase Bacillus subtilis

Storage Stability

EC Number Storage Stability Organism
1.4.3.19 -80°C, pH 7.5, both glycine oxidase and His-tagged glycine oxidase are stable for months Bacillus subtilis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.4.3.19 D-2-aminobutyrate + H2O + O2 as active as sarcosine Bacillus subtilis 2-oxobutyrate + H2O2
-
 ?
1.4.3.19 D-Ala + H2O + O2 about 115% of the activity with sarcosine Bacillus subtilis pyruvate + NH3 + H2O2
-
 ?
1.4.3.19 D-Arg + H2O + O2 about 35% of the activity with sarcosine Bacillus subtilis ? + H2O2
-
 ?
1.4.3.19 D-His + H2O + O2 about 25% of the activity with sarcosine Bacillus subtilis ? + H2O2
-
 ?
1.4.3.19 D-Ile + H2O + O2 about 30% of the activity with sarcosine Bacillus subtilis 3-methyl-2-oxopentanoate + H2O2
-
 ?
1.4.3.19 D-Leu + H2O + O2 about 35% of the activity with sarcosine Bacillus subtilis 4-methyl-2-oxopentanoate + H2O2
-
 ?
1.4.3.19 D-pipecolate + H2O + O2 about 85% of the activity with sarcosine Bacillus subtilis ? + H2O2
-
 ?
1.4.3.19 D-Pro + H2O + O2 about 120% of the activity with sarcosine Bacillus subtilis ? + H2O2
-
 ?
1.4.3.19 D-Val + H2O + O2 about 35% of the activity with sarcosine Bacillus subtilis 3-methyl-2-oxobutanoate + H2O2
-
 ?
1.4.3.19 glycine + H2O + O2 as active as sarcosine Bacillus subtilis glyoxylate + NH3 + H2O2
-
 ?
1.4.3.19 additional information the enzyme is strictly stereospecific as it only catalyzes the oxidation of the D-isomer Bacillus subtilis ?
-
 ?
1.4.3.19 N-ethylglycine + H2O + O2 about 65% of the activity with sarcosine Bacillus subtilis glyoxylate + ethylamine + H2O2
-
 ?
1.4.3.19 N-methyl-D-Ala + H2O + O2 about 110% of the activity with sarcosine Bacillus subtilis pyruvate + methylamine + H2O2
-
 ?
1.4.3.19 sarcosine + H2O + O2
-
 Bacillus subtilis glyoxylate + methylamine + H2O2
-
 ?

Subunits

EC Number Subunits Comment Organism
1.4.3.19 tetramer 4 * 46600, recombinant glycine oxidase, SDS-PAGE Bacillus subtilis
1.4.3.19 tetramer 4 * 49400, recombinant His-tagged chimeric glycine oxidase Bacillus subtilis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.4.3.19 additional information
-
 increase in activity with no evidence for any plateau or decrease up to 60°C, reaction with sarcosine Bacillus subtilis

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.4.3.19 15 60 increase in activity with no evidence for any plateau or decrease up to 60°C, reaction with sarcosine Bacillus subtilis

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.4.3.19 35
-
 pH 7.0, 30 min, stable up to Bacillus subtilis
1.4.3.19 45
-
 pH 7.0, 30 min, about 30% loss of activity Bacillus subtilis
1.4.3.19 46
-
 Tm-value, after 30 min Bacillus subtilis
1.4.3.19 50
-
 pH 7.0, 30 min, about 75% loss of activity Bacillus subtilis
1.4.3.19 60
-
 pH 7.0, 30 min, complete loss of activity Bacillus subtilis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.4.3.19 10
-
 reaction with sarcosine Bacillus subtilis

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.4.3.19 8 11 pH 8.0: about 55% of maximal activity, pH 11.0: about 55% of maximal activity, reaction with sarcosine Bacillus subtilis

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
1.4.3.19 7 8.5 25°C, 300 min, maximal stability Bacillus subtilis

Cofactor

EC Number Cofactor Comment Organism Structure
1.4.3.19 FAD tight binding of coenzyme to the protein moiety, addition of exogenous FAD or FMN to the assay mixture does not increase enzyme activity Bacillus subtilis

pI Value

EC Number Organism Comment pI Value Maximum pI Value
1.4.3.19 Bacillus subtilis theoretical value, His-tagged chimeric enzyme
-
 6
1.4.3.19 Bacillus subtilis theoretical value, wild-type enzyme
-
 6