Literature summary for 1.1.3.9 extracted from

Deacon, S.E.; McPherson, M.J.
Enhanced expression and purification of fungal galactose oxidase in Escherichia coli and use for analysis of a saturation mutagenesis library (2011), ChemBioChem, 12, 593-601.

Cloned(Commentary)

Cloned (Comment)	Organism
functional expression of wild-type and mutant enzymes in Escherichia coli strain BL21 Star (DE3) cytoplasm, best in strain BL21 Star (DE3) at 25°C, induction for 4-6 h is optimal	Fusarium graminearum

Protein Variants

Protein Variants	Comment	Organism
C383A	site-directed mutagenesis, the mutant shows 39% of wild-typ enzyme activity	Fusarium graminearum
C383D	site-directed mutagenesis, the mutant shows 26% of wild-typ enzyme activity	Fusarium graminearum
C383E	site-directed mutagenesis, the mutant shows 130% of wild-typ enzyme activity	Fusarium graminearum
C383F	site-directed mutagenesis, the mutant shows 8% of wild-typ enzyme activity	Fusarium graminearum
C383G	site-directed mutagenesis, the mutant shows 70% of wild-typ enzyme activity	Fusarium graminearum
C383H	site-directed mutagenesis, the mutant shows 35% of wild-typ enzyme activity	Fusarium graminearum
C383I	site-directed mutagenesis, the mutant shows 3% of wild-typ enzyme activity	Fusarium graminearum
C383K	site-directed mutagenesis, the mutant shows 115% of wild-typ enzyme activity	Fusarium graminearum
C383L	site-directed mutagenesis, the mutant shows 41% of wild-typ enzyme activity	Fusarium graminearum
C383M	site-directed mutagenesis, the mutant shows 75% of wild-typ enzyme activity	Fusarium graminearum
C383N	site-directed mutagenesis, the mutant shows 6% of wild-typ enzyme activity	Fusarium graminearum
C383P	site-directed mutagenesis, the mutant shows 18% of wild-typ enzyme activity	Fusarium graminearum
C383Q	site-directed mutagenesis, the mutant shows 13% of wild-typ enzyme activity	Fusarium graminearum
C383R	site-directed mutagenesis, the mutant shows 0.2% of wild-typ enzyme activity	Fusarium graminearum
C383S	site-directed mutagenesis, the mutant shows 160% of wild-typ enzyme activity	Fusarium graminearum
C383T	site-directed mutagenesis, the mutant shows 32% of wild-typ enzyme activity	Fusarium graminearum
C383V	site-directed mutagenesis, the mutant shows 4% of wild-typ enzyme activity	Fusarium graminearum
C383W	site-directed mutagenesis, the mutant shows 0.001% of wild-typ enzyme activity	Fusarium graminearum
C383Y	site-directed mutagenesis, the mutant cannt be isolated	Fusarium graminearum
G195E	site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme	Fusarium graminearum
M70V	site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme	Fusarium graminearum
additional information	introduction of silent mutations within codons 2-7 of the mature enzyme coding sequence to enhance enzyme translation and have combined these with other expression-enhancing mutations	Fusarium graminearum
N535D	site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme	Fusarium graminearum
V494A	site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme	Fusarium graminearum

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.0021	-	D-galactose	pH 7.0, 25°C, mutant C383E	Fusarium graminearum
0.0084	-	D-galactose	pH 7.0, 25°C, mutant C383D	Fusarium graminearum
0.03	-	D-galactose	pH 7.0, 25°C, mutant C383H	Fusarium graminearum
0.033	-	D-galactose	pH 7.0, 25°C, mutant C383M	Fusarium graminearum
0.034	-	D-galactose	pH 7.0, 25°C, mutant C383S	Fusarium graminearum
0.048	-	D-galactose	pH 7.0, 25°C, mutant C383K	Fusarium graminearum
0.054	-	D-galactose	pH 7.0, 25°C, wild-type enzyme	Fusarium graminearum
0.055	-	D-galactose	pH 7.0, 25°C, mutant C383L	Fusarium graminearum
0.06	-	D-galactose	pH 7.0, 25°C, mutant C383W	Fusarium graminearum
0.064	-	D-galactose	pH 7.0, 25°C, mutant C383Q	Fusarium graminearum
0.083	-	D-galactose	pH 7.0, 25°C, mutant C383G	Fusarium graminearum
0.13	-	D-galactose	pH 7.0, 25°C, mutant C383F	Fusarium graminearum
0.14	-	D-galactose	pH 7.0, 25°C, mutant C383P	Fusarium graminearum
0.16	-	D-galactose	pH 7.0, 25°C, mutant C383A	Fusarium graminearum
0.29	-	D-galactose	pH 7.0, 25°C, mutant C383R	Fusarium graminearum
0.39	-	D-galactose	pH 7.0, 25°C, mutant C383N	Fusarium graminearum
0.43	-	D-galactose	pH 7.0, 25°C, mutant C383V	Fusarium graminearum
0.51	-	D-galactose	pH 7.0, 25°C, mutant C383I	Fusarium graminearum
0.53	-	D-galactose	pH 7.0, 25°C, mutant C383T	Fusarium graminearum

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
D-galactose + O2	Fusarium graminearum	-	D-galacto-hexodialdose + H2O2	-	?

Organism

Organism	UniProt	Comment	Textmining
Fusarium graminearum	-	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
D-galactose + O2	-	Fusarium graminearum	D-galacto-hexodialdose + H2O2	-	?
additional information	the enzyme shows broad primary alcohol substrate specificity	Fusarium graminearum	?	-	?

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
25	-	assay at	Fusarium graminearum

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.011	-	D-galactose	pH 7.0, 25°C, mutant C383W	Fusarium graminearum
8.8	-	D-galactose	pH 7.0, 25°C, mutant C383R	Fusarium graminearum
170	-	D-galactose	pH 7.0, 25°C, mutant C383Q	Fusarium graminearum
190	-	D-galactose	pH 7.0, 25°C, mutant C383F	Fusarium graminearum
210	-	D-galactose	pH 7.0, 25°C, mutant C383H	Fusarium graminearum
260	-	D-galactose	pH 7.0, 25°C, mutant C383I	Fusarium graminearum
360	-	D-galactose	pH 7.0, 25°C, mutant C383V	Fusarium graminearum
410	-	D-galactose	pH 7.0, 25°C, mutant C383N	Fusarium graminearum
440	-	D-galactose	pH 7.0, 25°C, mutant C383D	Fusarium graminearum
450	-	D-galactose	pH 7.0, 25°C, mutant C383L	Fusarium graminearum
490	-	D-galactose	pH 7.0, 25°C, mutant C383P	Fusarium graminearum
510	-	D-galactose	pH 7.0, 25°C, mutant C383M	Fusarium graminearum
550	-	D-galactose	pH 7.0, 25°C, mutant C383E	Fusarium graminearum
1100	-	D-galactose	pH 7.0, 25°C, wild-type enzyme	Fusarium graminearum
1100	-	D-galactose	pH 7.0, 25°C, mutant C383G	Fusarium graminearum
1100	-	D-galactose	pH 7.0, 25°C, mutant C383K	Fusarium graminearum
1100	-	D-galactose	pH 7.0, 25°C, mutant C383S	Fusarium graminearum
1200	-	D-galactose	pH 7.0, 25°C, mutant C383A	Fusarium graminearum
3400	-	D-galactose	pH 7.0, 25°C, mutant C383T	Fusarium graminearum

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	-	assay at	Fusarium graminearum

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
0.00019	-	D-galactose	pH 7.0, 25°C, mutant C383W	Fusarium graminearum
0.03	-	D-galactose	pH 7.0, 25°C, mutant C383R	Fusarium graminearum
0.51	-	D-galactose	pH 7.0, 25°C, mutant C383I	Fusarium graminearum
0.83	-	D-galactose	pH 7.0, 25°C, mutant C383V	Fusarium graminearum
1.1	-	D-galactose	pH 7.0, 25°C, mutant C383N	Fusarium graminearum
1.5	-	D-galactose	pH 7.0, 25°C, mutant C383F	Fusarium graminearum
2.6	-	D-galactose	pH 7.0, 25°C, mutant C383Q	Fusarium graminearum
3.5	-	D-galactose	pH 7.0, 25°C, mutant C383P	Fusarium graminearum
5.2	-	D-galactose	pH 7.0, 25°C, mutant C383D	Fusarium graminearum
6.4	-	D-galactose	pH 7.0, 25°C, mutant C383T	Fusarium graminearum
7	-	D-galactose	pH 7.0, 25°C, mutant C383H	Fusarium graminearum
7.7	-	D-galactose	pH 7.0, 25°C, mutant C383A	Fusarium graminearum
8.2	-	D-galactose	pH 7.0, 25°C, mutant C383L	Fusarium graminearum
14	-	D-galactose	pH 7.0, 25°C, mutant C383G	Fusarium graminearum
15	-	D-galactose	pH 7.0, 25°C, mutant C383M	Fusarium graminearum
20	-	D-galactose	pH 7.0, 25°C, wild-type enzyme	Fusarium graminearum
23	-	D-galactose	pH 7.0, 25°C, mutant C383K	Fusarium graminearum
26	-	D-galactose	pH 7.0, 25°C, mutant C383E	Fusarium graminearum
32	-	D-galactose	pH 7.0, 25°C, mutant C383S	Fusarium graminearum