Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Bacillus subtilis |
Protein Variants | Comment | Organism |
---|---|---|
A54R | 20fold increase in Km for glyphosate | Bacillus subtilis |
G51R | 4000fold increase in the specificity constant for substrate glyphosate | Bacillus subtilis |
G51R/A54R | 5800fold increase in the specificity constant for substrate glyphosate | Bacillus subtilis |
G51S/A54R | 3100fold increase in the specificity constant for substrate glyphosate | Bacillus subtilis |
G51S/A54R/H244A | 210fold increase in catalytic activity and 15000fold increase in the specificity constant for substrate glyphosate. The alpha2-alpha3-loop assumes a different conformation, residue R54 may be the key residue in stabilizing glyphosate binding | Bacillus subtilis |
H244A | 2fold increase in expression yield | Bacillus subtilis |
Y241H | 100fold increase in the specificity constant for substrate glyphosate | Bacillus subtilis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.5 | - |
glyphosate | mutant G51S/A54R/H244A, pH 8.5, 25°C | Bacillus subtilis | |
0.7 | - |
glycine | wild-type, pH 8.5, 25°C | Bacillus subtilis | |
1 | - |
glyphosate | mutant G51R/A54R, pH 8.5, 25°C | Bacillus subtilis | |
1.3 | - |
glyphosate | mutant G51S/A54R, pH 8.5, 25°C | Bacillus subtilis | |
1.5 | - |
glycine | mutant H244A, pH 8.5, 25°C | Bacillus subtilis | |
4.4 | - |
glyphosate | mutant A54R, pH 8.5, 25°C | Bacillus subtilis | |
6.5 | - |
glyphosate | mutant G51R, pH 8.5, 25°C | Bacillus subtilis | |
28 | - |
glycine | mutant A54R, pH 8.5, 25°C | Bacillus subtilis | |
35 | - |
glycine | mutant G51S/A54R, pH 8.5, 25°C | Bacillus subtilis | |
38 | - |
glycine | mutant Y241H, pH 8.5, 25°C | Bacillus subtilis | |
41 | - |
glyphosate | mutant Y241H, pH 8.5, 25°C | Bacillus subtilis | |
53 | - |
glycine | mutant G51R, pH 8.5, 25°C | Bacillus subtilis | |
59 | - |
glycine | mutant G51R/A54R, pH 8.5, 25°C | Bacillus subtilis | |
78 | - |
glyphosate | mutant H244A, pH 8.5, 25°C | Bacillus subtilis | |
87 | - |
glyphosate | wild-type, pH 8.5, 25°C | Bacillus subtilis | |
105 | - |
glycine | mutant G51S/A54R/H244A, pH 8.5, 25°C | Bacillus subtilis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | O31616 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-Ala + H2O + O2 | 40% of the activity with sarcosine | Bacillus subtilis | ? + NH3 + H2O2 | - |
? | |
D-aminobutanoate + H2O + O2 | 30% of the activity with sarcosine | Bacillus subtilis | ? + H2O2 | - |
? | |
D-methionine + H2O + O2 | 25% of the activity with sarcosine | Bacillus subtilis | 4-(methylsulfanyl)-2-oxobutanoic acid + NH3 + H2O2 | - |
? | |
D-pipecolate + H2O + O2 | 70% of the activity with sarcosine | Bacillus subtilis | ? + H2O2 | - |
? | |
D-Pro + H2O + O2 | 110% of the activity with sarcosine | Bacillus subtilis | ? + NH3 + H2O2 | - |
? | |
glycine + H2O + O2 | 85% of the activity with sarcosine | Bacillus subtilis | glyoxylate + NH3 + H2O2 | - |
? | |
glycine-ethyl-ester + H2O + O2 | 90% of the activity with sarcosine | Bacillus subtilis | ? + H2O2 | - |
? | |
glyphosate + H2O + O2 | 110% of the activity with sarcosine | Bacillus subtilis | ? + H2O2 | - |
? | |
N-ethylglycine + H2O + O2 | 120% of the activity with sarcosine | Bacillus subtilis | glyoxylate + ethylamine + H2O2 | - |
? | |
N-methyl-D-Ala + H2O + O2 | 75% of the activity with sarcosine | Bacillus subtilis | ? + H2O2 | - |
? | |
sarcosine + H2O + O2 | - |
Bacillus subtilis | glyoxylate + methylamine + H2O2 | - |
? |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
34.7 | - |
melting temperature, mutant G51R/A54R, determined by following protein and flavin fluorescence | Bacillus subtilis |
41.3 | - |
melting temperature, mutant G51R, determined by following protein and flavin fluorescence | Bacillus subtilis |
44.8 | - |
melting temperature, mutant A54R, determined by following protein and flavin fluorescence | Bacillus subtilis |
45.3 | - |
melting temperature, mutant G51S/A54R, determined by following protein and flavin fluorescence | Bacillus subtilis |
45.7 | - |
melting temperature, mutant G51S/A54R/H244A, determined by following protein and flavin fluorescence | Bacillus subtilis |
56.9 | - |
melting temperature, wild-type, determined by following protein and flavin fluorescence | Bacillus subtilis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.35 | - |
glycine | mutant G51R, pH 8.5, 25°C | Bacillus subtilis | |
0.6 | - |
glycine | wild-type, pH 8.5, 25°C | Bacillus subtilis | |
0.63 | - |
glycine | mutant H244A, pH 8.5, 25°C | Bacillus subtilis | |
0.7 | - |
glycine | mutant G51R/A54R, pH 8.5, 25°C | Bacillus subtilis | |
0.7 | - |
glyphosate | mutant G51R/A54R, pH 8.5, 25°C | Bacillus subtilis | |
0.77 | - |
glyphosate | mutant H244A, pH 8.5, 25°C | Bacillus subtilis | |
0.91 | - |
glyphosate | wild-type, pH 8.5, 25°C | Bacillus subtilis | |
0.91 | - |
glycine | mutant G51S/A54R, pH 8.5, 25°C | Bacillus subtilis | |
0.91 | - |
glycine | mutant Y241H, pH 8.5, 25°C | Bacillus subtilis | |
1.05 | - |
glyphosate | mutant G51S/A54R, pH 8.5, 25°C | Bacillus subtilis | |
1.05 | - |
glyphosate | mutant G51S/A54R/H244A, pH 8.5, 25°C | Bacillus subtilis | |
1.2 | - |
glycine | mutant A54R, pH 8.5, 25°C | Bacillus subtilis | |
1.3 | - |
glyphosate | mutant Y241H, pH 8.5, 25°C | Bacillus subtilis | |
1.5 | - |
glyphosate | mutant A54R, pH 8.5, 25°C | Bacillus subtilis | |
1.5 | - |
glycine | mutant G51S/A54R/H244A, pH 8.5, 25°C | Bacillus subtilis | |
1.8 | - |
glyphosate | mutant G51R, pH 8.5, 25°C | Bacillus subtilis |