Table 1. Amino acids and positions within characterized AA5_2 sequences that are implicated in catalysis and substrate preference.

F. graminearum					Corresponding amino acid in
Catagory	Position	Amino acid	Reported function	Reference(s)	CgrRaOx	CgrAlcOx	PruAA5_2
Catalysis	194	F	π-π interaction with F227 and W290	[39]	F	W	Y
	228	C	C228-Y272 redox cofactor	[3,5,40]	C	C	C
	272	Y	C228-Y272 redox cofactor, copper ligand	[3,5,40]	Y	Y	Y
	441	F	π-π stacking interaction with Y495	[3]	F	F	F
	464	F	π-π stacking interaction with Y495	[39]	F	F	F
	495	Y	Catalytic tyrosine, copper ligand	[3,5]	Y	Y	Y
	496	H	Copper ligand	[3,5]	H	H	H
	581	H	Copper ligand	[3,5]	H	H	H
Substrate preference	290	W	π-π stacking interaction with Y272 and F194, hydrogen bonding with D-galactose	[3,5,16,34,40–42]	Y	F	W
	326	Qa	Hydrogen bonding with R330	[34,41]	A	G	D
	329	Y	Hydrogen bonding with D-galactose and R330	[34]	W	M	Y
	330	R	Bidentate hydrogen bonds with D-galactose	[34,37,41]	R	F	R
	405	Y	Hydrogen bonding with Y495	[37]	Y	Y	Y
	406	Qa	Hydrogen bonding with D-galactose	[34,37]	S	T	E
Catalytic efficiency	383	Ca	C383S increases Vmax on D-galactose by 1.75x and lowers KM by 3.6x	[35,36]	C	C	S
	436	Ya	Y436H increases Vmax on D-galactose by 2x	[35]	K	Y	A
	494	Va	V494A increases Vmax on D-galactose by 1.75x	[35,36,41]	N	N	V

Amino acid positions correspond to the archetypal galactose oxidase from Fusarium graminearum.

^aAmino acids not fully conserved among AA5_2 sequences from other Fusarium species (Fig A in S1 File).