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Literature summary for 1.4.3.19 extracted from

  • Boselli, A.; Rosini, E.; Marcone, G.L.; Sacchi, S.; Motteran, L.; Pilone, M.S.; Pollegioni, L.; Molla, G.
    Glycine oxidase from Bacillus subtilis: role of histidine 244 and methionine 261 (2007), Biochimie, 89, 1372-1380.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli strain BL21(DE3)pLysS Bacillus subtilis

Protein Variants

Protein Variants Comment Organism
H244F shows increased kcat value for glycine compared to the wild type enzyme, the mutation does not affect the expression of glycine oxidase and the physicochemical properties of bound FAD Bacillus subtilis
H244N shows increased kcat value for glycine compared to the wild type enzyme, the mutation does not affect the expression of glycine oxidase and the physicochemical properties of bound FAD Bacillus subtilis
H244Q shows increased kcat value compared to the wild type enzyme, the mutation does not affect the expression of glycine oxidase and the physicochemical properties of bound FAD Bacillus subtilis
M261H shows decreased kcat value for glycine compared to the wild type enzyme, the mutation does not affect the expression of glycine oxidase and the physicochemical properties of bound FAD Bacillus subtilis
M261Y shows decreased kcat value for glycine compared to the wild type enzyme, the mutation does not affect the expression of glycine oxidase and the physicochemical properties of bound FAD Bacillus subtilis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.6
-
glycine apparent value for mutant enzyme H244F, at 25°C and pH 8.5 Bacillus subtilis
0.7
-
glycine apparent value for wild type enzyme, at 25°C and pH 8.5 Bacillus subtilis
0.7
-
sarcosine apparent value for wild type enzyme, at 25°C and pH 8.5 Bacillus subtilis
0.8
-
glycine apparent value for mutant enzyme H244Q, at 25°C and pH 8.5 Bacillus subtilis
1.4
-
sarcosine apparent value for mutant enzyme H244F, at 25°C and pH 8.5 Bacillus subtilis
1.9
-
glycine apparent value for mutant enzyme H244N, at 25°C and pH 8.5 Bacillus subtilis
2.1
-
sarcosine apparent value for mutant enzyme H244N, at 25°C and pH 8.5 Bacillus subtilis
2.8
-
sarcosine apparent value for mutant enzyme H244Q, at 25°C and pH 8.5 Bacillus subtilis
4 10 D-alanine apparent value for mutant enzyme M261Y, at 25°C and pH 8.5 Bacillus subtilis
11
-
sarcosine apparent value for mutant enzyme M261H, at 25°C and pH 8.5 Bacillus subtilis
11
-
sarcosine apparent value for mutant enzyme M261Y, at 25°C and pH 8.5 Bacillus subtilis
17
-
D-proline apparent value for wild type enzyme, at 25°C and pH 8.5 Bacillus subtilis
18
-
glycine apparent value for mutant enzyme M261Y, at 25°C and pH 8.5 Bacillus subtilis
26
-
D-proline apparent value for mutant enzyme H244F, at 25°C and pH 8.5 Bacillus subtilis
26
-
D-proline apparent value for mutant enzyme H244N, at 25°C and pH 8.5 Bacillus subtilis
27
-
D-proline apparent value for mutant enzyme H244Q, at 25°C and pH 8.5 Bacillus subtilis
28
-
glycine apparent value for mutant enzyme M261H, at 25°C and pH 8.5 Bacillus subtilis
280
-
D-proline apparent value for mutant enzyme M261H, at 25°C and pH 8.5 Bacillus subtilis
285
-
D-proline apparent value for mutant enzyme M261Y, at 25°C and pH 8.5 Bacillus subtilis
350
-
D-alanine apparent value for mutant enzyme H244Q, at 25°C and pH 8.5 Bacillus subtilis
420
-
D-alanine apparent value for mutant enzyme H244N, at 25°C and pH 8.5 Bacillus subtilis
490
-
D-alanine apparent value for wild type enzyme, at 25°C and pH 8.5 Bacillus subtilis
530
-
D-alanine apparent value for mutant enzyme H244F, at 25°C and pH 8.5 Bacillus subtilis
2000
-
D-alanine Km above 2000 mM, apparent value for mutant enzyme M261H, at 25°C and pH 8.5 Bacillus subtilis

Organism

Organism UniProt Comment Textmining
Bacillus subtilis
-
-
-

Purification (Commentary)

Purification (Comment) Organism
HiTrap chelating affinity chromatography 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
-
?
sarcosine + H2O + O2
-
Bacillus subtilis glyoxylate + methylamine + H2O2
-
?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.3
-
D-proline apparent value for mutant enzyme H244N, at 25°C and pH 8.5 Bacillus subtilis
0.3
-
glycine apparent value for mutant enzyme M261Y, at 25°C and pH 8.5 Bacillus subtilis
0.33
-
D-alanine kcat above 0.33 1/s, apparent value for mutant enzyme M261H, at 25°C and pH 8.5 Bacillus subtilis
0.38
-
D-proline apparent value for mutant enzyme M261Y, at 25°C and pH 8.5 Bacillus subtilis
0.47
-
D-alanine apparent value for mutant enzyme M261Y, at 25°C and pH 8.5 Bacillus subtilis
0.47
-
sarcosine apparent value for mutant enzyme M261Y, at 25°C and pH 8.5 Bacillus subtilis
0.5
-
D-proline apparent value for wild type enzyme, at 25°C and pH 8.5 Bacillus subtilis
0.55
-
glycine apparent value for mutant enzyme M261H, at 25°C and pH 8.5 Bacillus subtilis
0.6
-
glycine apparent value for wild type enzyme, at 25°C and pH 8.5 Bacillus subtilis
0.6
-
sarcosine apparent value for wild type enzyme, at 25°C and pH 8.5 Bacillus subtilis
0.62
-
D-proline apparent value for mutant enzyme M261H, at 25°C and pH 8.5 Bacillus subtilis
0.73
-
sarcosine apparent value for mutant enzyme M261H, at 25°C and pH 8.5 Bacillus subtilis
0.75
-
D-alanine apparent value for mutant enzyme H244N, at 25°C and pH 8.5 Bacillus subtilis
0.78
-
glycine apparent value for mutant enzyme H244F, at 25°C and pH 8.5 Bacillus subtilis
0.92
-
D-alanine apparent value for wild type enzyme, at 25°C and pH 8.5 Bacillus subtilis
1.08
-
sarcosine apparent value for mutant enzyme H244F, at 25°C and pH 8.5 Bacillus subtilis
1.08
-
D-alanine apparent value for mutant enzyme H244Q, at 25°C and pH 8.5 Bacillus subtilis
1.12
-
sarcosine apparent value for mutant enzyme H244Q, at 25°C and pH 8.5 Bacillus subtilis
1.2
-
glycine apparent value for mutant enzyme H244Q, at 25°C and pH 8.5 Bacillus subtilis
1.23
-
D-proline apparent value for mutant enzyme H244Q, at 25°C and pH 8.5 Bacillus subtilis
1.3
-
glycine apparent value for mutant enzyme H244N, at 25°C and pH 8.5 Bacillus subtilis
1.33
-
sarcosine apparent value for mutant enzyme H244N, at 25°C and pH 8.5 Bacillus subtilis
1.35
-
D-proline apparent value for mutant enzyme H244F, at 25°C and pH 8.5 Bacillus subtilis
1.5
-
D-alanine apparent value for mutant enzyme H244F, at 25°C and pH 8.5 Bacillus subtilis

Cofactor

Cofactor Comment Organism Structure
FAD contains one molecule of non-covalently bound FAD per subunit Bacillus subtilis