Literature summary for 3.2.1.25 extracted from

Kaper, T.; van Heusden, H.H.; van Loo, B.; Vasella, A.; van der Oost, J.; de Vos, W.M.
Substrate specificity engineering of beta-mannosidase and beta-glucosidase from Pyrococcus by exchange of unique active site residues (2002), Biochemistry, 41, 4147-4155.

Search for more literature for 3.2.1.25

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli	Pyrococcus horikoshii

Protein Variants

Protein Variants	Comment	Organism
D206N	in contrast to the wild-type enzyme which shows a catalytic efficiency for 4-nitrophenyl beta-D-mannopyranoside that is 5fold higher than that for 4-nitrophenyl beta-D-galactopyranoside and 4fold higher than that for 4-nitrophenyl beta-D-glucopyranoside, the mutant enzyme shows highest catalytic efficiency towards 4-nitrophenyl beta-D-glucopyranoside, 11fold higher than that for 4-nitrophenyl beta-D-galactopyranoside and 60fold higher than that for 4-nitrophenyl beta-D-mannopyranoside	Pyrococcus horikoshii
Q77R	in contrast to the wild-type enzyme which shows a catalytic efficiency for 4-nitrophenyl beta-D-mannopyranoside that is 5fold higher than that for 4-nitrophenyl beta-D-galactopyranoside and 4fold higher than that for 4-nitrophenyl beta-D-glucopyranoside, the mutant enzyme shows highest catalytic efficiency towards 4-nitrophenyl beta-D-glucopyranoside, 2fold higher than that for 4-nitrophenyl beta-D-galactopyranoside and 1.1fold higher than that for 4-nitrophenyl beta-D-mannopyranoside	Pyrococcus horikoshii
Q77R/D206N	in contrast to the wild-type enzyme which shows a catalytic efficiency for 4-nitrophenyl beta-D-mannopyranoside that is 5fold higher than that for 4-nitrophenyl beta-D-galactopyranoside and 4fold higher than that for 4-nitrophenyl beta-D-glucopyranoside, the mutant enzyme shows highest catalytic efficiency towards 4-nitrophenyl beta-D-glucopyranoside, 8fold higher than that for 4-nitrophenyl beta-D-galactopyranoside and 1.4fold higher than that for 4-nitrophenyl beta-D-mannopyranoside	Pyrococcus horikoshii

Inhibitors

Inhibitors	Comment	Organism	Structure
(5R,6R,7S,8S)-5-(hydroxymethyl)-2-phenyl-5,6,7,8-tetrahydroimidazol[1,2-a]pyridine-6,7,8-triol	-	Pyrococcus horikoshii

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.3	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, mutant enzyme D206N	Pyrococcus horikoshii
0.44	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, wild-type enzyme	Pyrococcus horikoshii
0.68	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme Q77R/D206N	Pyrococcus horikoshii
1.1	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme Q77R	Pyrococcus horikoshii
3.2	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme D206N	Pyrococcus horikoshii
7.3	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, wild-type enzyme	Pyrococcus horikoshii
7.8	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, mutant enzyme D206N	Pyrococcus horikoshii
13.5	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, wild-type enzyme	Pyrococcus horikoshii
16.2	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, mutant enzyme Q77R	Pyrococcus horikoshii
16.8	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, mutant enzyme Q77R	Pyrococcus horikoshii
30.4	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, mutant enzyme Q77R/D206N	Pyrococcus horikoshii
36.1	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, mutant enzyme Q77R/D206N	Pyrococcus horikoshii

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
56457	-	4 * 56457, calculated from sequence	Pyrococcus horikoshii
56500	-	4 * 56500, SDS-PAGE	Pyrococcus horikoshii
250000	-	gel filtration	Pyrococcus horikoshii

Organism

Organism	UniProt	Comment	Textmining
Pyrococcus horikoshii	O58237	-	-
Pyrococcus horikoshii DSM 12428	O58237	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Pyrococcus horikoshii

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
4-nitrophenyl beta-D-galactopyranoside + H2O	catalytic efficiency (kcat/Km) of the wild-type enzyme for 4-nitrophenyl beta-D-galactopyranoside is 20% compared to catalytic efficiency (kcat/Km) of the wild-type enzyme for 4-nitrophenyl-beta-D-mannopyranoside	Pyrococcus horikoshii	4-nitrophenol + beta-D-galactose	-	?
4-nitrophenyl beta-D-galactopyranoside + H2O	catalytic efficiency (kcat/Km) of the wild-type enzyme for 4-nitrophenyl beta-D-galactopyranoside is 20% compared to catalytic efficiency (kcat/Km) of the wild-type enzyme for 4-nitrophenyl-beta-D-mannopyranoside	Pyrococcus horikoshii DSM 12428	4-nitrophenol + beta-D-galactose	-	?
4-nitrophenyl beta-D-glucopyranoside + H2O	catalytic efficiency (kcat/Km) of the wild-type enzyme for 4-nitrophenyl beta-D-glucopyranoside is 26% compared to catalytic efficiency (kcat/Km) of the wild-type enzyme for 4-nitrophenyl beta-D-mannopyranoside. The catalytic efficiency (kcat/Km) of the mutant enzymes Q77R, D206N and Q77R/D206N for 4-nitrophenyl beta-D-glucopyranoside is higher then that for 4-nitrophenyl beta-D-galactopyranoside and 4-nitrophenyl-beta-D-mannopyranoside	Pyrococcus horikoshii	4-nitrophenol + beta-D-glucose	-	?
4-nitrophenyl beta-D-glucopyranoside + H2O	catalytic efficiency (kcat/Km) of the wild-type enzyme for 4-nitrophenyl beta-D-glucopyranoside is 26% compared to catalytic efficiency (kcat/Km) of the wild-type enzyme for 4-nitrophenyl beta-D-mannopyranoside. The catalytic efficiency (kcat/Km) of the mutant enzymes Q77R, D206N and Q77R/D206N for 4-nitrophenyl beta-D-glucopyranoside is higher then that for 4-nitrophenyl beta-D-galactopyranoside and 4-nitrophenyl-beta-D-mannopyranoside	Pyrococcus horikoshii DSM 12428	4-nitrophenol + beta-D-glucose	-	?
4-nitrophenyl beta-D-mannopyranoside + H2O	-	Pyrococcus horikoshii	4-nitrophenol + beta-D-mannose	-	?
4-nitrophenyl beta-D-mannopyranoside + H2O	-	Pyrococcus horikoshii DSM 12428	4-nitrophenol + beta-D-mannose	-	?

Subunits

Subunits	Comment	Organism
tetramer	4 * 56500, SDS-PAGE	Pyrococcus horikoshii
tetramer	4 * 56457, calculated from sequence	Pyrococcus horikoshii

Synonyms

Synonyms	Comment	Organism
BglB	-	Pyrococcus horikoshii
PH0501	locus name	Pyrococcus horikoshii

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
101	-	melting temperature is 100.7°C	Pyrococcus horikoshii
102	-	half-life: 27 min	Pyrococcus horikoshii

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
1.6	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme Q77R	Pyrococcus horikoshii
4.2	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, wild-type enzyme	Pyrococcus horikoshii
6.8	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme Q77R/D206N	Pyrococcus horikoshii
13.5	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, wild-type enzyme	Pyrococcus horikoshii
14.2	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, mutant enzyme Q77R	Pyrococcus horikoshii
25.5	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, mutant enzyme Q77R	Pyrococcus horikoshii
34.4	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, wild-type enzyme	Pyrococcus horikoshii
45	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme D206N	Pyrococcus horikoshii
66.5	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, mutant enzyme Q77R/D206N	Pyrococcus horikoshii
252	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, mutant enzyme D206N	Pyrococcus horikoshii
439	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, mutant enzyme Q77R/D206N	Pyrococcus horikoshii
571	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, mutant enzyme D206N	Pyrococcus horikoshii

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
4.75	-	-	Pyrococcus horikoshii

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.000014	-	(5R,6R,7S,8S)-5-(hydroxymethyl)-2-phenyl-5,6,7,8-tetrahydroimidazol[1,2-a]pyridine-6,7,8-triol	pH 5.0, 90°C, mutant enzyme D206N	Pyrococcus horikoshii
0.02	-	(5R,6R,7S,8S)-5-(hydroxymethyl)-2-phenyl-5,6,7,8-tetrahydroimidazol[1,2-a]pyridine-6,7,8-triol	pH 5.0, 90°C, mutant enzyme Q77R/D206N	Pyrococcus horikoshii
0.1	-	(5R,6R,7S,8S)-5-(hydroxymethyl)-2-phenyl-5,6,7,8-tetrahydroimidazol[1,2-a]pyridine-6,7,8-triol	pH 5.0, 90°C, wild-type enzyme	Pyrococcus horikoshii
0.11	-	(5R,6R,7S,8S)-5-(hydroxymethyl)-2-phenyl-5,6,7,8-tetrahydroimidazol[1,2-a]pyridine-6,7,8-triol	pH 5.0, 90°C, mutant enzyme Q77R	Pyrococcus horikoshii

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.85	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, mutant enzyme Q77R	Pyrococcus horikoshii
1.4	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme Q77R	Pyrococcus horikoshii
1.6	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, mutant enzyme Q77R	Pyrococcus horikoshii
1.8	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, mutant enzyme Q77R/D206N	Pyrococcus horikoshii
1.9	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, wild-type enzyme	Pyrococcus horikoshii
2.58	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, wild-type enzyme	Pyrococcus horikoshii
9.8	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, wild-type enzyme	Pyrococcus horikoshii
9.9	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme Q77R/D206N	Pyrococcus horikoshii
14.4	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, mutant enzyme Q77R/D206N	Pyrococcus horikoshii
14.7	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme D206N	Pyrococcus horikoshii
75.8	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, mutant enzyme D206N	Pyrococcus horikoshii
870	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, mutant enzyme D206N	Pyrococcus horikoshii

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Release 2023.1 (January 2023)