1.1.3.9: galactose oxidase

This is an abbreviated version!
For detailed information about galactose oxidase, go to the full flat file.

Word Map on EC 1.1.3.9

Reaction

D-galactose
+
O2
=
D-galacto-hexodialdose
+
H2O2

Synonyms

AOd, beta-galactose oxidase, D-galactose oxidase, FgrGalOx, galactose 6-oxidase, galactose oxidase, GalOx, GAO, GAOA, GAOX, GO, GOase

ECTree

     1 Oxidoreductases
         1.1 Acting on the CH-OH group of donors
             1.1.3 With oxygen as acceptor
                1.1.3.9 galactose oxidase

Engineering

Engineering on EC 1.1.3.9 - galactose oxidase

Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C383W
-
site-directed mutagenesis, the mutant shows 0.001% of wild-typ enzyme activity
C383R
-
site-directed mutagenesis, the mutant shows 0.2% of wild-typ enzyme activity
C383S
-
site-directed mutagenesis, the mutant shows 160% of wild-typ enzyme activity
W290H
kcat/KM for D-galactose is 1180fold lower than wild-type value
W290G
the activity of Trp290 mutants of FgrGalOx show a dramatic loss of oxidative capacity compared to wild-type, which is correlated to significantly higher Km values for the natural substrate galactose with the FgrGalOx W290G/F mutants, presumably because of the loss of a hydrogen-bonding interaction between W290 and a remote hydroxyl group of the substrate. Trp290 in FgrGalOx is implicated in stabilizing the radical form of the Cys-Tyr cofactor, although substitution with either Phe or Gly also stabilizes the tyrosine radical with retention of catalytic activity, while other substitutions were detrimental to the enzyme
C383T
-
site-directed mutagenesis, the mutant shows 32% of wild-typ enzyme activity
C383V
-
site-directed mutagenesis, the mutant shows 4% of wild-typ enzyme activity
C383Q
-
site-directed mutagenesis, the mutant shows 13% of wild-typ enzyme activity
C383Y
-
site-directed mutagenesis, the mutant cannt be isolated
G195E
-
site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme
M70V
-
site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme
W290G
kcat/KM for D-galactose is 6370fold lower than wild-type value
V494A
-
site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme
C383G
-
site-directed mutagenesis, the mutant shows 70% of wild-typ enzyme activity
W290F
N535D
-
site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme
C383A
-
site-directed mutagenesis, the mutant shows 39% of wild-typ enzyme activity
W290F
-
structure PDB ID 2EIC, comparison to the wild-type enzyme. The activity of Trp290 mutants of FgrGalOx show a dramatic loss of oxidative capacity compared to wild-type, which is correlated to significantly higher Km values for the natural substrate galactose with the FgrGalOx W290G/F mutants, presumably because of the loss of a hydrogen-bonding interaction between W290 and a remote hydroxyl group of the substrate. Trp290 in FgrGalOx is implicated in stabilizing the radical form of the Cys-Tyr cofactor, although substitution with either Phe or Gly also stabilizes the tyrosine radical with retention of catalytic activity, while other substitutions were detrimental to the enzyme
C383D
-
site-directed mutagenesis, the mutant shows 26% of wild-typ enzyme activity
C383E
-
site-directed mutagenesis, the mutant shows 130% of wild-typ enzyme activity
C383F
-
site-directed mutagenesis, the mutant shows 8% of wild-typ enzyme activity
C383H
-
site-directed mutagenesis, the mutant shows 35% of wild-typ enzyme activity
C383I
-
site-directed mutagenesis, the mutant shows 3% of wild-typ enzyme activity
C383K
-
site-directed mutagenesis, the mutant shows 115% of wild-typ enzyme activity
C383L
-
site-directed mutagenesis, the mutant shows 41% of wild-typ enzyme activity
C383M
-
site-directed mutagenesis, the mutant shows 75% of wild-typ enzyme activity
C383N
-
site-directed mutagenesis, the mutant shows 6% of wild-typ enzyme activity
C383P
-
site-directed mutagenesis, the mutant shows 18% of wild-typ enzyme activity
V494A
-
slight decrease in Km-value, increase in kcat-value
S10P/M70 V/P136/G195E/V494A/N535D
-
random mutagenesis, the enzyme mutant shows improved levels of recombinant expression of a more active and stable enzyme in Escherichia coli without any change in substrate range compared to the wild-type enzyme
R330K/W290F/Q406E/Y405F
-
the mutant shows 136fold increased activity with D-fructose, and increased activity with mannose and N-acetylglucosamine compared to the wild-type enzyme
Y436H/V494A
-
slight decrease in kcat-value of 1-methyl-alpha-D-galactose, slight increase in kcat-value of D-galactose
R330K
-
the mutant shows increased activity with D-fructose compared to the wild-type enzyme
W290F/R330K/Q406T
-
random mutagenesis, the mutant shows improved activity toward Glc compared to the wild-type enzyme
W290H
-
the mutant is highly activates by phosphate
Y405F/Q406E
-
random mutagenesis, the mutant shows altered substrate specificity with several substrates compared to the wild-type enzyme
Y405F/Q406Y
-
random mutagenesis, the mutant shows altered substrate specificity with several substrates compared to the wild-type enzyme
Y436A
-
Km- and kcat-values similar to wild type
Y436A/V494A
-
decrease in Km-value, increase in kcat-value
Y436H
-
slight decrease in Km-value, decrease in kcat-value
C383S/Y436H/V494A
-
decrease in Km-value, increase in kcat-value
C383/Y436H
-
decrease in Km-value, increase in kcat-value
C383A
-
slight decrease in Km-value, decrease in kcat-value
C383H/V494A
-
decrease in Km-value, decrease in kcat-value
C383S
-
decrease in Km-value
C383S/Y436A
-
decrease in Km-value, decrease in kcat-value of 1-methyl-alpha-D-galactose, increase in kcat-value of D-galactose
P463V
-
random mutagenesis, the mutant shows altered substrate specificity with several substrates compared to the wild-type enzyme
P463I
-
random mutagenesis, the mutant shows altered substrate specificity with several substrates compared to the wild-type enzyme
W290H
-
the mutant is highly activates by phosphate
-
V494A
-
slight decrease in Km-value, increase in kcat-value
-
Y436H
-
slight decrease in Km-value, decrease in kcat-value
-
C383S
-
decrease in Km-value
-
C383A
-
slight decrease in Km-value, decrease in kcat-value
-
Y436A
-
Km- and kcat-values similar to wild type
-
C228G
Y272F
-
not capable of binding copper
W290H
-
analysis from X-ray structure
additional information