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Literature summary for 1.4.3.19 extracted from

  • Martinez-Martinez, I.; Navarro-Fernandez, J.; Lozada-Ramirez, J.D.; Garcia-Carmona, F.; Sanchez-Ferrer, A.
    Maximization of production of his-tagged glycine oxidase and its M261 mutant proteins (2006), Biotechnol. Prog., 22, 647-652.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
cloning of the glycine oxidase gene from Bacillus subtilis ATCC 6633 into the Rosetta Escherichia coli strain which contains the pRARE plasmid, inserting of the gene into the pET28a expression vector Bacillus subtilis

Protein Variants

Protein Variants Comment Organism
M261H decreased in Km compared with the wild type Bacillus subtilis
M261Y decreased in Km compared with the wild type Bacillus subtilis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.18
-
glycine M261Y mutant Bacillus subtilis
0.23
-
sarcosine M261Y mutant Bacillus subtilis
0.29
-
glycine-ethyl-ester M261Y mutant Bacillus subtilis
0.56
-
glycine M261 wild type Bacillus subtilis
0.57
-
sarcosine M261 wild type Bacillus subtilis
0.58
-
sarcosine M261H mutant Bacillus subtilis
0.71
-
N-ethyl-glycine M261Y mutant Bacillus subtilis
0.97
-
glycine M261H mutant Bacillus subtilis
1.57
-
glycine-ethyl-ester M261H mutant Bacillus subtilis
1.78
-
glycine-ethyl-ester M261 wild type Bacillus subtilis
2.06
-
D-proline M261Y mutant Bacillus subtilis
2.07
-
N-ethyl-glycine M261H mutant Bacillus subtilis
2.8
-
N-ethyl-glycine M261 wild type Bacillus subtilis
3.91
-
D-proline M261H mutant Bacillus subtilis
8.9
-
D-proline M261 wild type Bacillus subtilis
121.9
-
D-alanine M261Y mutant Bacillus subtilis
155.6
-
D-alanine M261H mutant Bacillus subtilis
315
-
D-alanine M261 wild type Bacillus subtilis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
42000
-
monomer, determined by SDS-PAGE Bacillus subtilis
172000
-
homotetramer, determined by gel filtration Bacillus subtilis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
D-alanine + H2O + O2 Bacillus subtilis
-
pyruvate + NH3 + H2O2
-
?
D-proline + H2O + O2 Bacillus subtilis
-
?
-
?
glycine + H2O + O2 Bacillus subtilis
-
glyoxylate + NH3 + H2O2
-
?
glycine-ethyl-ester + H2O + O2 Bacillus subtilis
-
?
-
?
N-ethylglycine + H2O + O2 Bacillus subtilis
-
glyoxylate + ethylamine + H2O2
-
?
sarcosine + H2O + O2 Bacillus subtilis
-
glyoxylate + methylamine + H2O2
-
?

Organism

Organism UniProt Comment Textmining
Bacillus subtilis
-
-
-

Purification (Commentary)

Purification (Comment) Organism
using ultrafiltation and a HiTrap chelating affinity column Bacillus subtilis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.19
-
crude extract Bacillus subtilis
0.21
-
purification step ultrafiltration Bacillus subtilis
1.2
-
purification step HiTrap chelating chromatography Bacillus subtilis

Storage Stability

Storage Stability Organism
-80°C, 75 mM sodium diphosphate buffer, pH 8.5, 10% glycerol Bacillus subtilis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-alanine + H2O + O2
-
Bacillus subtilis pyruvate + NH3 + H2O2
-
?
D-proline + H2O + O2
-
Bacillus subtilis ?
-
?
glycine + H2O + O2
-
Bacillus subtilis glyoxylate + NH3 + H2O2
-
?
glycine-ethyl-ester + H2O + O2
-
Bacillus subtilis ?
-
?
N-ethylglycine + H2O + O2
-
Bacillus subtilis glyoxylate + ethylamine + H2O2
-
?
sarcosine + H2O + O2
-
Bacillus subtilis glyoxylate + methylamine + H2O2
-
?

Subunits

Subunits Comment Organism
homotetramer
-
Bacillus subtilis

Synonyms

Synonyms Comment Organism
glycin oxidase
-
Bacillus subtilis
GOX
-
Bacillus subtilis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
activity assay Bacillus subtilis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
activity assay Bacillus subtilis

Cofactor

Cofactor Comment Organism Structure
FAD
-
Bacillus subtilis