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Literature summary for 1.4.3.19 extracted from

  • Pedotti, M.; Rosini, E.; Molla, G.; Moschetti, T.; Savino, C.; Vallone, B.; Pollegioni, L.
    Glyphosate resistance by engineering the flavoenzyme glycine oxidase (2009), J. Biol. Chem., 284, 36415-36423.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.4.3.19

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Bacillus subtilis

Protein Variants

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 [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

Organism UniProt Comment Textmining
Bacillus subtilis O31616
-
-

Substrates and Products (Substrate)

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 [°C]

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 [1/s]

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