Literature summary for 3.2.1.73 extracted from

Cheng, Y.S.; Huang, C.H.; Chen, C.C.; Huang, T.Y.; Ko, T.P.; Huang, J.W.; Wu, T.H.; Liu, J.R.; Guo, R.T.
Structural and mutagenetic analyses of a 1,3-1,4-beta-glucanase from Paecilomyces thermophila (2014), Biochim. Biophys. Acta, 1844, 366-373.

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified wild-type PtLic16A and inactive mutant E113A in ligand-free form and in complex with the ligands cellobiose, cellotetraose and glucotriose, X-ray diffraction structure determination and analysis at 1.80-2.25 A resolution. Native PtLic16A crystals belong to the space group C2221 and contain four polypeptide chains in an asymmetric unit. The E113A mutant crystals belong to a different space group of C2 and have two protein molecules in an asymmetric unit. The third E113A crystal belongs to space group P21, with four molecules in an asymmetric unit	Paecilomyces sp. 'thermophila'
purified wild-type PtLic16A and inactive mutant E113A in ligand-free form and in complex with the ligands cellobiose, cellotetraose and glucotriose, X-ray diffraction structure determination and analysis at 1.80-2.25 A resolution. Native PtLic16A crystals belong to the space group C2221 and contain four polypeptide chains in an asymmetric unit. The E113A mutant crystals belong to a different space group of C2 and have two protein molecules in an asymmetric unit. The third E113A crystal belongs to space group P21, with four molecules in an asymmetric unit	Paecilomyces sp. (in: Eurotiomycetes)

Crystallization (Comment)

Organism

purified wild-type PtLic16A and inactive mutant E113A in ligand-free form and in complex with the ligands cellobiose, cellotetraose and glucotriose, X-ray diffraction structure determination and analysis at 1.80-2.25 A resolution. Native PtLic16A crystals belong to the space group C2221 and contain four polypeptide chains in an asymmetric unit. The E113A mutant crystals belong to a different space group of C2 and have two protein molecules in an asymmetric unit. The third E113A crystal belongs to space group P21, with four molecules in an asymmetric unit

Paecilomyces sp. 'thermophila'

Paecilomyces sp. (in: Eurotiomycetes)

Protein Variants

Protein Variants	Comment	Organism
E113A	inactive mutant. In the structure of E113A/1,3-1,4-beta-glucotriose complex, the sugar bound to the -1 subsite adopts an intermediate-like (alpha-anomeric) configuration	Paecilomyces sp. 'thermophila'
E113A	inactive mutant. In the structure of E113A/1,3-1,4-beta-glucotriose complex, the sugar bound to the -1 subsite adopts an intermediate-like (alpha-anomeric) configuration	Paecilomyces sp. (in: Eurotiomycetes)
W108A	site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme	Paecilomyces sp. 'thermophila'
W108A	site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme	Paecilomyces sp. (in: Eurotiomycetes)
W108F	site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme	Paecilomyces sp. 'thermophila'
W108F	site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme	Paecilomyces sp. (in: Eurotiomycetes)
W108Y	site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme	Paecilomyces sp. 'thermophila'
W108Y	site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme	Paecilomyces sp. (in: Eurotiomycetes)
W253A	site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme	Paecilomyces sp. 'thermophila'
W253A	site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme	Paecilomyces sp. (in: Eurotiomycetes)
W253F	site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme	Paecilomyces sp. 'thermophila'
W253F	site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme	Paecilomyces sp. (in: Eurotiomycetes)
W253Y	site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme	Paecilomyces sp. 'thermophila'
W253Y	site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme	Paecilomyces sp. (in: Eurotiomycetes)

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
additional information	Paecilomyces sp. 'thermophila'	the enzyme specifically cleaves the beta-1,4-glycosidic bonds adjacent to beta-1,3-linkages in polysaccharides such as beta-D-glucan or lichenan	?	-	?
additional information	Paecilomyces sp. (in: Eurotiomycetes)	the enzyme specifically cleaves the beta-1,4-glycosidic bonds adjacent to beta-1,3-linkages in polysaccharides such as beta-D-glucan or lichenan	?	-	?
additional information	Paecilomyces sp. (in: Eurotiomycetes) J18	the enzyme specifically cleaves the beta-1,4-glycosidic bonds adjacent to beta-1,3-linkages in polysaccharides such as beta-D-glucan or lichenan	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Paecilomyces sp. 'thermophila'	-	-	-
Paecilomyces sp. (in: Eurotiomycetes)	E0XN39	-	-
Paecilomyces sp. (in: Eurotiomycetes) J18	E0XN39	-	-

Reaction

Reaction	Comment	Organism	Reaction ID
beta-D-glucopyranosyl-(1-3)-beta-D-glucopyranosyl-(1-3)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-3)-beta-D-glucopyranose + 4 H2O = 5 beta-D-glucopyranose	the enzyme might carry out the hydrolysis via retaining mechanism with E113 and E118 serving as the nucleophile and general acid/base, respectively,catalytic mechanism, overview	Paecilomyces sp. 'thermophila'	a
beta-D-glucopyranosyl-(1-3)-beta-D-glucopyranosyl-(1-3)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-3)-beta-D-glucopyranose + 4 H2O = 5 beta-D-glucopyranose	the enzyme might carry out the hydrolysis via retaining mechanism with E113 and E118 serving as the nucleophile and general acid/base, respectively,catalytic mechanism, overview	Paecilomyces sp. (in: Eurotiomycetes)	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
barley beta-glucan + H2O	-	Paecilomyces sp. 'thermophila'	?	-	?
barley beta-glucan + H2O	-	Paecilomyces sp. (in: Eurotiomycetes)	?	-	?
barley beta-glucan + H2O	-	Paecilomyces sp. (in: Eurotiomycetes) J18	?	-	?
lichenan + H2O	-	Paecilomyces sp. 'thermophila'	?	-	?
lichenan + H2O	-	Paecilomyces sp. (in: Eurotiomycetes)	?	-	?
lichenan + H2O	-	Paecilomyces sp. (in: Eurotiomycetes) J18	?	-	?
additional information	the enzyme specifically cleaves the beta-1,4-glycosidic bonds adjacent to beta-1,3-linkages in polysaccharides such as beta-D-glucan or lichenan	Paecilomyces sp. 'thermophila'	?	-	?
additional information	the enzyme specifically cleaves the beta-1,4-glycosidic bonds adjacent to beta-1,3-linkages in polysaccharides such as beta-D-glucan or lichenan	Paecilomyces sp. (in: Eurotiomycetes)	?	-	?
additional information	the enzyme specifically cleaves the beta-1,4-glycosidic bonds adjacent to beta-1,3-linkages in polysaccharides such as beta-D-glucan or lichenan. Computer modeling of active-site bound oligosaccharides, overview	Paecilomyces sp. 'thermophila'	?	-	?
additional information	the enzyme specifically cleaves the beta-1,4-glycosidic bonds adjacent to beta-1,3-linkages in polysaccharides such as beta-D-glucan or lichenan. Computer modeling of active-site bound oligosaccharides, overview	Paecilomyces sp. (in: Eurotiomycetes)	?	-	?
additional information	the enzyme specifically cleaves the beta-1,4-glycosidic bonds adjacent to beta-1,3-linkages in polysaccharides such as beta-D-glucan or lichenan	Paecilomyces sp. (in: Eurotiomycetes) J18	?	-	?
additional information	the enzyme specifically cleaves the beta-1,4-glycosidic bonds adjacent to beta-1,3-linkages in polysaccharides such as beta-D-glucan or lichenan. Computer modeling of active-site bound oligosaccharides, overview	Paecilomyces sp. (in: Eurotiomycetes) J18	?	-	?

Subunits

Subunits	Comment	Organism
monomer	-	Paecilomyces sp. 'thermophila'
monomer	-	Paecilomyces sp. (in: Eurotiomycetes)

Synonyms

Synonyms	Comment	Organism
1,3-1,4-beta-glucanase	-	Paecilomyces sp. 'thermophila'
1,3-1,4-beta-glucanase	-	Paecilomyces sp. (in: Eurotiomycetes)
Lichenase	-	Paecilomyces sp. 'thermophila'
Lichenase	-	Paecilomyces sp. (in: Eurotiomycetes)
PtLic16A	-	Paecilomyces sp. 'thermophila'
PtLic16A	-	Paecilomyces sp. (in: Eurotiomycetes)

General Information

General Information	Comment	Organism
additional information	the enzyme adopts a typical beta-jellyroll fold with a curved surface and the concave face forms an extended ligand binding cleft. Both catalytic residues E113 and E118 are embedded in the strand beta9. The ligand binding cleft is formed by the inner beta-sheets, four associated loops and one alpha-helix, which connect the strands beta2 to beta3 (residues 20-29), beta8 to beta9 (105-112), beta11 to beta12 (150164) and beta16 to alpha6 (250-260). The outer beta-sheets did not involve in the active site formation but might play a structural role in overall folding. Existence of a detailed interaction network in the active site and the intermediate-like configuration, overview	Paecilomyces sp. 'thermophila'
additional information	the enzyme adopts a typical beta-jellyroll fold with a curved surface and the concave face forms an extended ligand binding cleft. Both catalytic residues E113 and E118 are embedded in the strand beta9. The ligand binding cleft is formed by the inner beta-sheets, four associated loops and one alpha-helix, which connect the strands beta2 to beta3 (residues 20-29), beta8 to beta9 (105-112), beta11 to beta12 (150164) and beta16 to alpha6 (250-260). The outer beta-sheets did not involve in the active site formation but might play a structural role in overall folding. Existence of a detailed interaction network in the active site and the intermediate-like configuration, overview	Paecilomyces sp. (in: Eurotiomycetes)