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Literature summary for 1.4.3.19 extracted from

  • Rosini, E.; Piubelli, L.; Molla, G.; Frattini, L.; Valentino, M.; Varriale, A.; DAuria, S.; Pollegioni, L.
    Novel biosensors based on optimized glycine oxidase (2014), FEBS J., 281, 3460-3472 .
    View publication on PubMed
Search for more literature for 1.4.3.19

Cloned(Commentary)

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

Protein Variants

Protein Variants Comment Organism
A54E the mutant shows reduced catalytic efficiency with glycine compared to the wild type enzyme Bacillus subtilis
A54R the mutant shows reduced catalytic efficiency with glycine compared to the wild type enzyme Bacillus subtilis
A54R/H244K the mutant shows reduced catalytic efficiency with glycine compared to the wild type enzyme Bacillus subtilis
A54R/H244K/M261R the mutant shows reduced catalytic efficiency with glycine compared to the wild type enzyme Bacillus subtilis
G51H the mutant shows reduced catalytic efficiency with glycine compared to the wild type enzyme Bacillus subtilis
G51Q the mutant shows reduced catalytic efficiency with glycine compared to the wild type enzyme Bacillus subtilis
H244A the mutant shows reduced catalytic efficiency with glycine compared to the wild type enzyme Bacillus subtilis
H244K the mutant shows increased catalytic efficiency with glycine compared to the wild type enzyme Bacillus subtilis
H244K/M261R the variant shows a 5.4fold increase in maximal activity on glycine compared to the wild type enzyme Bacillus subtilis
H244N the mutant shows reduced catalytic efficiency with glycine compared to the wild type enzyme Bacillus subtilis
H244Q the mutant shows increased catalytic efficiency with glycine compared to the wild type enzyme Bacillus subtilis
H244R the mutant shows increased catalytic efficiency with glycine compared to the wild type enzyme Bacillus subtilis
M261I the mutant shows reduced catalytic efficiency with glycine compared to the wild type enzyme Bacillus subtilis
M261R the mutant shows reduced catalytic efficiency with glycine compared to the wild type enzyme Bacillus subtilis
M49I the mutant shows increased catalytic efficiency with glycine compared to the wild type enzyme Bacillus subtilis
M49L the mutant shows reduced catalytic efficiency with glycine compared to the wild type enzyme Bacillus subtilis
M49L/A54R/H244K the mutant shows reduced catalytic efficiency with glycine compared to the wild type enzyme Bacillus subtilis
M49T the mutant shows reduced catalytic efficiency with glycine compared to the wild type enzyme Bacillus subtilis
Y246W the mutant shows reduced catalytic efficiency with glycine compared to the wild type enzyme Bacillus subtilis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.14
-
 glycine mutant enzyme H244K, at pH 8.5 and 25°C Bacillus subtilis
0.24
-
 glycine mutant enzyme H244R, at pH 8.5 and 25°C Bacillus subtilis
0.3
-
 glycine mutant enzyme H244K/M261R, at pH 8.5 and 25°C Bacillus subtilis
0.31
-
 glycine mutant enzyme M49I, at pH 8.5 and 25°C Bacillus subtilis
0.7
-
 glycine wild type enzyme, at pH 8.5 and 25°C Bacillus subtilis
0.7
-
 sarcosine wild type enzyme, at pH 8.5 and 25°C Bacillus subtilis
0.9
-
 glycine mutant enzyme H244Q, at pH 8.5 and 25°C Bacillus subtilis
1.35
-
 glycine mutant enzyme M49L, at pH 8.5 and 25°C Bacillus subtilis
1.47
-
 glycine mutant enzyme M49T, at pH 8.5 and 25°C Bacillus subtilis
1.5
-
 glycine mutant enzyme H244A, at pH 8.5 and 25°C Bacillus subtilis
1.5
-
 glycine mutant enzyme M261I, at pH 8.5 and 25°C Bacillus subtilis
1.9
-
 glycine mutant enzyme H244N, at pH 8.5 and 25°C Bacillus subtilis
3.5
-
 glycine mutant enzyme M261R, at pH 8.5 and 25°C Bacillus subtilis
10
-
 glycine mutant enzyme A54E, at pH 8.5 and 25°C Bacillus subtilis
14.6
-
 glycine mutant enzyme G51Q, at pH 8.5 and 25°C Bacillus subtilis
17.2
-
 glycine mutant enzyme Y246W, at pH 8.5 and 25°C Bacillus subtilis
18.8
-
 glycine mutant enzyme M49L/A54R/H244K, at pH 8.5 and 25°C Bacillus subtilis
28
-
 glycine mutant enzyme A54R, at pH 8.5 and 25°C Bacillus subtilis
35.7
-
 glycine mutant enzyme G51H, at pH 8.5 and 25°C Bacillus subtilis
40.1
-
 glycine mutant enzyme A54R/H244K, at pH 8.5 and 25°C Bacillus subtilis
45.9
-
 glycine mutant enzyme A54R/H244K/M261R, at pH 8.5 and 25°C Bacillus subtilis

Organism

Organism UniProt Comment Textmining
Bacillus subtilis O31616
-
-
Bacillus subtilis 168 O31616
-
-

Purification (Commentary)

Purification (Comment) Organism
HiTrap chelating column chromatography Bacillus subtilis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
glycine + H2O + O2
-
 Bacillus subtilis glyoxylate + NH3 + H2O2
-
 ?
glycine + H2O + O2
-
 Bacillus subtilis 168 glyoxylate + NH3 + H2O2
-
 ?
sarcosine + H2O + O2
-
 Bacillus subtilis glyoxylate + methylamine + H2O2
-
 ?
sarcosine + H2O + O2
-
 Bacillus subtilis 168 glyoxylate + methylamine + H2O2
-
 ?

Subunits

Subunits Comment Organism
homotetramer 4 * 47000, SDS-PAGE Bacillus subtilis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
57
-
 melting temperature Bacillus subtilis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.04
-
 glycine mutant enzyme M49T, at pH 8.5 and 25°C Bacillus subtilis
0.35
-
 glycine mutant enzyme A54E, at pH 8.5 and 25°C Bacillus subtilis
0.36
-
 glycine mutant enzyme M49I, at pH 8.5 and 25°C Bacillus subtilis
0.43
-
 glycine mutant enzyme M49L, at pH 8.5 and 25°C Bacillus subtilis
0.6
-
 glycine mutant enzyme M49L/A54R/H244K, at pH 8.5 and 25°C Bacillus subtilis
0.6
-
 glycine wild type enzyme, at pH 8.5 and 25°C Bacillus subtilis
0.6
-
 sarcosine wild type enzyme, at pH 8.5 and 25°C Bacillus subtilis
0.63
-
 glycine mutant enzyme H244A, at pH 8.5 and 25°C Bacillus subtilis
0.64
-
 glycine mutant enzyme M261I, at pH 8.5 and 25°C Bacillus subtilis
0.74
-
 glycine mutant enzyme G51Q, at pH 8.5 and 25°C Bacillus subtilis
0.77
-
 glycine mutant enzyme G51H, at pH 8.5 and 25°C Bacillus subtilis
1
-
 glycine mutant enzyme M261R, at pH 8.5 and 25°C Bacillus subtilis
1.2
-
 glycine mutant enzyme A54R, at pH 8.5 and 25°C Bacillus subtilis
1.2
-
 glycine mutant enzyme H244K/M261R, at pH 8.5 and 25°C Bacillus subtilis
1.21
-
 glycine mutant enzyme H244Q, at pH 8.5 and 25°C Bacillus subtilis
1.21
-
 glycine mutant enzyme Y246W, at pH 8.5 and 25°C Bacillus subtilis
1.3
-
 glycine mutant enzyme H244N, at pH 8.5 and 25°C Bacillus subtilis
1.35
-
 glycine mutant enzyme H244K, at pH 8.5 and 25°C Bacillus subtilis
1.49
-
 glycine mutant enzyme H244R, at pH 8.5 and 25°C Bacillus subtilis
1.61
-
 glycine mutant enzyme A54R/H244K/M261R, at pH 8.5 and 25°C Bacillus subtilis
3.2
-
 glycine mutant enzyme A54R/H244K, at pH 8.5 and 25°C Bacillus subtilis

Cofactor

Cofactor Comment Organism Structure
FAD
-
 Bacillus subtilis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.02
-
 glycine mutant enzyme G51H, at pH 8.5 and 25°C Bacillus subtilis
0.03
-
 glycine mutant enzyme M49L/A54R/H244K, at pH 8.5 and 25°C Bacillus subtilis
0.03
-
 glycine mutant enzyme M49T, at pH 8.5 and 25°C Bacillus subtilis
0.04
-
 glycine mutant enzyme A54E, at pH 8.5 and 25°C Bacillus subtilis
0.04
-
 glycine mutant enzyme A54R, at pH 8.5 and 25°C Bacillus subtilis
0.04
-
 glycine mutant enzyme A54R/H244K/M261R, at pH 8.5 and 25°C Bacillus subtilis
0.05
-
 glycine mutant enzyme G51Q, at pH 8.5 and 25°C Bacillus subtilis
0.07
-
 glycine mutant enzyme Y246W, at pH 8.5 and 25°C Bacillus subtilis
0.11
-
 glycine mutant enzyme A54R/H244K, at pH 8.5 and 25°C Bacillus subtilis
0.3
-
 glycine mutant enzyme M261R, at pH 8.5 and 25°C Bacillus subtilis
0.32
-
 glycine mutant enzyme M49L, at pH 8.5 and 25°C Bacillus subtilis
0.42
-
 glycine mutant enzyme H244A, at pH 8.5 and 25°C Bacillus subtilis
0.43
-
 glycine mutant enzyme M261I, at pH 8.5 and 25°C Bacillus subtilis
0.68
-
 glycine mutant enzyme H244N, at pH 8.5 and 25°C Bacillus subtilis
0.86
-
 glycine wild type enzyme, at pH 8.5 and 25°C Bacillus subtilis
0.86
-
 sarcosine wild type enzyme, at pH 8.5 and 25°C Bacillus subtilis
1.17
-
 glycine mutant enzyme M49I, at pH 8.5 and 25°C Bacillus subtilis
1.34
-
 glycine mutant enzyme H244Q, at pH 8.5 and 25°C Bacillus subtilis
4
-
 glycine mutant enzyme H244K/M261R, at pH 8.5 and 25°C Bacillus subtilis
6.22
-
 glycine mutant enzyme H244R, at pH 8.5 and 25°C Bacillus subtilis
9.65
-
 glycine mutant enzyme H244K, at pH 8.5 and 25°C Bacillus subtilis