Literature summary for 3.2.1.91 extracted from

Momeni, M.; Ubhayasekera, W.; Sandgren, M.; Stahlberg, J.; Hansson, H.
Structural insights into the inhibition of cellobiohydrolase Cel7A by xylo-oligosaccharides (2015), FEBS J., 282, 2167-2177 .

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified catalytic modules of wild-type enzyme Cel7A and mutants E212Q and E217Q in complex with xylooligosaccharides, hanging drop vapor diffusion method, mixing of equal volumes of 6 mg/ml protein in 10 mM sodium acetate, pH 5.0, with reservoir solution containing 20% PEG 5000 monomethyl ether, 0.1 M MES, pH 6.0, 10 mM CoCl2, and 12.5% glycerol, microseeding, 1-2 days, X-ray diffraction structure determination and analysis at 1.42-1.89 A resolution, structure modelling. Each structure model contains all 434 amino acid residues of the HjeCel7A catalytic module, with an N-terminal pyroglutamate residue and an N-acetyl glucosamine residue bound to Asn270	Trichoderma reesei

Crystallization (Comment)

Organism

purified catalytic modules of wild-type enzyme Cel7A and mutants E212Q and E217Q in complex with xylooligosaccharides, hanging drop vapor diffusion method, mixing of equal volumes of 6 mg/ml protein in 10 mM sodium acetate, pH 5.0, with reservoir solution containing 20% PEG 5000 monomethyl ether, 0.1 M MES, pH 6.0, 10 mM CoCl2, and 12.5% glycerol, microseeding, 1-2 days, X-ray diffraction structure determination and analysis at 1.42-1.89 A resolution, structure modelling. Each structure model contains all 434 amino acid residues of the HjeCel7A catalytic module, with an N-terminal pyroglutamate residue and an N-acetyl glucosamine residue bound to Asn270

Trichoderma reesei

Protein Variants

Protein Variants	Comment	Organism
E212Q	site-directed mutagenesis, catalytic residue, inactive mutant	Trichoderma reesei
E217Q	site-directed mutagenesis, catalytic residue, inactive mutant	Trichoderma reesei

Inhibitors

Inhibitors	Comment	Organism
additional information	xylan and xylooligosaccharides (XOS) play a key role in inhibition of cellobiohydrolases of glycoside hydrolase family 7. Analysis of the binding mode at the entrance of the substrate-binding tunnel of the enzyme, in which each xylose residue is shifted about 2.4 A towards the catalytic center compared with binding of cello-oligosaccharides. Partial occupancy of two consecutive xylose residues at subsites -2 and -1 suggests an alternative binding mode for XOS in the vicinity of the catalytic center. The -1 xylosyl unit exhibits an open aldehyde conformation in one of the structures and a ring-closed pyranoside in another complex. Complementary inhibition studies with 4-nitrophenyl lactoside as substrate indicate mixed inhibition rather than pure competitive inhibition. Inhibitor binding structure analysis with wild-type and mutant Cel7A enzymes, detailed overview. No inhibition by xylose	Trichoderma reesei
xylan	from birchwood	Trichoderma reesei
xylobiose	mixed-type inhibition	Trichoderma reesei
xylopentaose	mixed-type inhibition	Trichoderma reesei
xylotriose	mixed-type inhibition	Trichoderma reesei

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	Michaelis-Menten kinetics	Trichoderma reesei
1.6	-	4-nitrophenyl lactoside	pH 5.0, 37°C, recombinant wild-type enzyme	Trichoderma reesei

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
extracellular	-	Trichoderma reesei	-	-

Organism

Organism	UniProt	Comment	Textmining
Trichoderma reesei	G0RVK1	i.e. Hypocrea jecorina	-

Source Tissue

Source Tissue	Comment	Organism	Textmining
additional information	the major enzyme, cellobiohydrolase Cel7A, constitutes nearly half of the total protein in the secretome	Trichoderma reesei	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
4-nitrophenyl lactoside + H2O	-	Trichoderma reesei	4-nitrophenol + lactose	-	?

Synonyms

Synonyms	Comment	Organism
Cel7A	-	Trichoderma reesei
cellobiohydrolase	-	Trichoderma reesei

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Trichoderma reesei

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
5	-	assay at	Trichoderma reesei

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism
additional information	-	additional information	Michaelis-Menten kinetics indicating mixed-type inhibition	Trichoderma reesei
9.7	-	xylobiose	pH 5.0, 37°C, inhibition of recombinant wild-type enzyme	Trichoderma reesei
14	-	xylopentaose	pH 5.0, 37°C, inhibition of recombinant wild-type enzyme	Trichoderma reesei
29	-	xylotriose	pH 5.0, 37°C, inhibition of recombinant wild-type enzyme	Trichoderma reesei

General Information

General Information	Comment	Organism
evolution	the enzyme belongs to the glycoside hydrolase family 7, GH7	Trichoderma reesei