Literature summary 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.

Cloned(Commentary)

EC Number	Cloned (Comment)	Organism
1.4.3.19	expression in Escherichia coli	Bacillus subtilis

Protein Variants

EC Number	Protein Variants	Comment	Organism
1.4.3.19	A54R	20fold increase in Km for glyphosate	Bacillus subtilis
1.4.3.19	G51R	4000fold increase in the specificity constant for substrate glyphosate	Bacillus subtilis
1.4.3.19	G51R/A54R	5800fold increase in the specificity constant for substrate glyphosate	Bacillus subtilis
1.4.3.19	G51S/A54R	3100fold increase in the specificity constant for substrate glyphosate	Bacillus subtilis
1.4.3.19	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
1.4.3.19	H244A	2fold increase in expression yield	Bacillus subtilis
1.4.3.19	Y241H	100fold increase in the specificity constant for substrate glyphosate	Bacillus subtilis

KM Value [mM]

EC Number	KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
1.4.3.19	0.5	-	glyphosate	mutant G51S/A54R/H244A, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	0.7	-	glycine	wild-type, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	1	-	glyphosate	mutant G51R/A54R, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	1.3	-	glyphosate	mutant G51S/A54R, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	1.5	-	glycine	mutant H244A, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	4.4	-	glyphosate	mutant A54R, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	6.5	-	glyphosate	mutant G51R, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	28	-	glycine	mutant A54R, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	35	-	glycine	mutant G51S/A54R, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	38	-	glycine	mutant Y241H, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	41	-	glyphosate	mutant Y241H, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	53	-	glycine	mutant G51R, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	59	-	glycine	mutant G51R/A54R, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	78	-	glyphosate	mutant H244A, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	87	-	glyphosate	wild-type, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	105	-	glycine	mutant G51S/A54R/H244A, pH 8.5, 25°C	Bacillus subtilis

Organism

EC Number	Organism	UniProt	Comment	Textmining
1.4.3.19	Bacillus subtilis	O31616	-	-

Substrates and Products (Substrate)

EC Number	Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
1.4.3.19	D-Ala + H2O + O2	40% of the activity with sarcosine	Bacillus subtilis	? + NH3 + H2O2	-	?
1.4.3.19	D-aminobutanoate + H2O + O2	30% of the activity with sarcosine	Bacillus subtilis	? + H2O2	-	?
1.4.3.19	D-methionine + H2O + O2	25% of the activity with sarcosine	Bacillus subtilis	4-(methylsulfanyl)-2-oxobutanoic acid + NH3 + H2O2	-	?
1.4.3.19	D-pipecolate + H2O + O2	70% of the activity with sarcosine	Bacillus subtilis	? + H2O2	-	?
1.4.3.19	D-Pro + H2O + O2	110% of the activity with sarcosine	Bacillus subtilis	? + NH3 + H2O2	-	?
1.4.3.19	glycine + H2O + O2	85% of the activity with sarcosine	Bacillus subtilis	glyoxylate + NH3 + H2O2	-	?
1.4.3.19	glycine-ethyl-ester + H2O + O2	90% of the activity with sarcosine	Bacillus subtilis	? + H2O2	-	?
1.4.3.19	glyphosate + H2O + O2	110% of the activity with sarcosine	Bacillus subtilis	? + H2O2	-	?
1.4.3.19	N-ethylglycine + H2O + O2	120% of the activity with sarcosine	Bacillus subtilis	glyoxylate + ethylamine + H2O2	-	?
1.4.3.19	N-methyl-D-Ala + H2O + O2	75% of the activity with sarcosine	Bacillus subtilis	? + H2O2	-	?
1.4.3.19	sarcosine + H2O + O2	-	Bacillus subtilis	glyoxylate + methylamine + H2O2	-	?

Temperature Stability [°C]

EC Number	Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
1.4.3.19	34.7	-	melting temperature, mutant G51R/A54R, determined by following protein and flavin fluorescence	Bacillus subtilis
1.4.3.19	41.3	-	melting temperature, mutant G51R, determined by following protein and flavin fluorescence	Bacillus subtilis
1.4.3.19	44.8	-	melting temperature, mutant A54R, determined by following protein and flavin fluorescence	Bacillus subtilis
1.4.3.19	45.3	-	melting temperature, mutant G51S/A54R, determined by following protein and flavin fluorescence	Bacillus subtilis
1.4.3.19	45.7	-	melting temperature, mutant G51S/A54R/H244A, determined by following protein and flavin fluorescence	Bacillus subtilis
1.4.3.19	56.9	-	melting temperature, wild-type, determined by following protein and flavin fluorescence	Bacillus subtilis

Turnover Number [1/s]

EC Number	Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
1.4.3.19	0.35	-	glycine	mutant G51R, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	0.6	-	glycine	wild-type, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	0.63	-	glycine	mutant H244A, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	0.7	-	glycine	mutant G51R/A54R, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	0.7	-	glyphosate	mutant G51R/A54R, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	0.77	-	glyphosate	mutant H244A, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	0.91	-	glyphosate	wild-type, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	0.91	-	glycine	mutant G51S/A54R, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	0.91	-	glycine	mutant Y241H, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	1.05	-	glyphosate	mutant G51S/A54R, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	1.05	-	glyphosate	mutant G51S/A54R/H244A, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	1.2	-	glycine	mutant A54R, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	1.3	-	glyphosate	mutant Y241H, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	1.5	-	glyphosate	mutant A54R, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	1.5	-	glycine	mutant G51S/A54R/H244A, pH 8.5, 25°C	Bacillus subtilis
1.4.3.19	1.8	-	glyphosate	mutant G51R, pH 8.5, 25°C	Bacillus subtilis

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Release 2023.1 (January 2023)