TABLE 2.

Carbohydrate-active enzymes detected at protein level that may take part in anaerobic filter paper and woodchip degradation and interaction between microbial species^f

Substratea	Predicted enzyme activity	CAZy annotation	MAGs/fungi	Location
Cellulose	Cellobiohydrolase, reducing end-acting	GH7 GH48;CBM2	Spoth2 Bin.005, Bin.009	WBR E
	Cellobiohydrolase, non-reducing end-acting	CBM2;GH6	Bin.009	E
	Endo-β−1,4-glucanasea	GH5_2 GH6;CBM2 GH44a (CBM30;)CBM11;CBM11;GH51_3 GH8b	Bin.064 Bin.005, Bin.006 Bin.038 Bin.063, Bin.064 Bin.063, Bin.064	E E E E E
	Endo-β−1,4-glucanase, processive	CBM4;GH9 GH9;CBM2 GH9	Bin.005, Bin.009 Bin.006, Bin.009 Bin.038, Bin.064	E E E
	Cellodextrin phosphorylase	GH94	Bin.005, Bin.009, Bin.037	E
	β-Glucosidase	GH3c GH30_1	Bin.012, Bin.030, Bin.031, Bin.038 Bin.038	E, WBR E
	Pyrroloquinoline quinone-dependent (glucose) dehydrogenase	AA12	Neucr2	WBR
	Lytic polysaccharide monooxygenase	AA10;CBM2	Bin.009	E
	Cellobiose dehydrogenase with an iron reductase domain	AA3_1;AA8	Thite2, Aspfu1	WBR
	FAD-dependent gluco-oligosaccharide oxidase	AA7	Bin.099	E
	Glucose oxidase / Glucose dehydrogenased	AA3_2
	Pyranose 2-oxidase	AA3_4	Bin.099	E
Xyloglucan	(Xyloglucan specific) endo-β-1,4-glucanase	GH44a	Bin.038	E
	α-Xylosidase	GH31_4	Bin.028, Bin.038	E
	β-Galactosidase	GH2	Bin.038, Bin.089	E
Pectin	Pectate lyase	PL1_7 PL9_3	Aspfu1 Bin.013	WBR WBR
	Pectate disaccharide-lyase/oligogalacturonate lyase	PL22	Bin.111	E
	α-Rhamnosidase	CBM67;GH78	Bin.038	E
	Pectin methylesterase	CE8	Bin.076	WBR
Galacto(gluco)mannan	β-Mannosidase	GH2	Bin.038	E
	α-Galactosidase	GH36	Bin.038	E
Xylan	Endo-β-1,4-xylanase	GH10 GH8	Colhig2, Neucr2 Bin.063, Bin.064,	WBR E
	Acetylxylan esterase/feruloyl esterasee	CE1	Spoth2, Bin.038, Bin.111	WBR, E
	α−1,2-(4-O-methyl)-glucuronidase	GH115	Bin.139	WBR
Lignin–carbohydrate complexes	(4-O-methyl)-D-glucuronate–lignin esterase	CE15	Thite2	WBR
Lignin	Peroxidase	AA2	Bin.013	WBR
	Aryl alcohol oxidase/aryl alcohol dehydrogenased	AA3_2	Bin.099, Bin.112, Thite2, Colhig2	E, WBR
	Alcohol oxidase	AA3_3	Colhig2; Fusfu1	WBR
	NAD(P)H:quinone oxidoreductase (not secreted)	AA6	Bin.100	E
Starch	Amylase, 1,4-α-glucan branching enzyme	CBM48;GH13_9	Bin.005, Bin.009	E
	α−1,4-Glucan phosphorylase	GT35	Bin.009	E
Trehalose	α,α-Trehalase	GH37	Colhig2	WBR
	Trehalose-6-phosphate synthase	GT20	Bin.009	E
Bacterial cell wall	Peptidoglycan transferase, monofunctional Peptidoglycan transferase, bifunctional	GT51 GT51; penicillin-binding transpeptidase	Bin.111 Bin.099, Bin.101, Bin.111	E E
	Peptidoglycan lytic transglycosylase	GH102	Bin.111	E
	N-Acetylgalactosaminidase	GH109	Bin.038, Bin.090	E
Galactosaminogalactan (fungal exopolysaccharide)	Endo-α−1,4-N-acetylgalactosaminidase	GH135	Bin.099, Bin.100	E
Chitin (fungal cell wall)	Chitinase	GH18	Bin.111	E
	β-Hexosaminidase	GH3	Bin.112	E
Protein glycosylation (N-glycans in plants)	α-Mannosidase, non-reducing end-acting	GH92	Bin.037	E
Glycosaminoglycans, glycoproteins	β-N-Acetylhexosaminidase	GH20 GH179	Bin.037, Bin.038 Bin.009	E E
Other	UDP-3-O-acyl-N-acetylglucosamine deacetylase	CE11	Bin.038, Bin.111	E
	Polysaccharide synthase	GT2	Bin.061, Bin.111	E

^a Some enzyme classes may also exhibit activity towards other substrates, including xyloglucans, xylans, and mannans.

^b Some enzymes in the GH8 family are endo-β-1,3-xylanases or reducing end-active xylose-releasing exo-oligoxylanases.

^c Some enzymes in the GH3 family are β-xylosidases.

^d Some AA3_2 enzymes listed under aryl alcohol oxidases or dehydrogenases may be glucose oxidases or dehydrogenases.

^e Some CE1 deacetylases may also deacetylate other acetylated polysaccharides, such as mannans.

^f The table organizes the detected enzymes based on their predicted activity and target substrate and shows the CAZy modules identified in the domain structure and the MAGs/fungi expressing such proteins. Location indicates whether the enzymes were detected in the WBR (wood chips) or in the enrichment flasks (E; primarily filter paper), or both. Abbreviations: Aspfu1, Aspergillus fumigatus; Colhig2, Colletotrichum higginsianum; Fusfu1, Fusarium fujikuroi; Neucr2, Neurospora crassa; Spoth2, Thermothelomyces thermophilus; Thite2, Thermothielavioides terrestris; AA, auxiliary activity; CBM, carbohydrate-binding module; GH, glycoside hydrolase; GT, glycoside transferase; PL – polysaccharide lyase; CE, carbohydrate esterase.