Table 2.

Screening substrates.

Function	Enzyme	Substrate	Substrate concentration	Detection (nm)	Assay type
Cellulose degradation	Cellulase	AZCL-HE-Cellulose	0.1% (w/v)	590	Liquid
	Cellobiohydrolase	4-MUB-β-D-Cellobiose	1 mM	365/450	Liquid
	β-Glucosidase	PNP-β-D-Glucoside	1 mM	410	Liquid
Hemicellulose degradation	Xylanase	AZCL-Xylan	0.1% (w/v)	590	Liquid
	β-Xylosidase	4-MUB-β-D-Xyloside		365/450	Liquid
Starch degradation	α-Amylase	Starch Azure	0.5% (w/v)	590	Liquid
	α-Glucosidase	4-MUB-α-D-Glucoside	1 mM	365/450	Liquid
Chitin degradation	Chitinase	Chitin azure	0.1% (w/v)	590	Liquid
	N-AcetyL-β-D-glucosaminidase	4-MUB-β-N-acetylglucosamine	1 mM	365/450	Liquid
Lignin degradation	Polyphenol oxidase	L-dihydroxyphenylalanine	5 mM	480	Liquid
	Laccase	Syringaldazine	40 μM	530	Liquid
PO4 mineralization	Phosphatase	5-Bromo-4-chloro-3-indolyl phosphate	0.1 mM	Visual	Solid
Protein turnover	Protease	Skim milk	2%	Visual	Solid

In order to allow simultaneous detection of multiple activities, azurin cross-linked (AZCL), p-nitrophenyl (PNP) and 4-methylumbelliferyl (MUB) labeled carbohydrates were pooled on the same assay plate per polymer type, e.g., AZCL Xylan and -MUB- β-D-Xyloside were pooled for screening for activities involved in hemicellulose degradation.