Literature summary for 3.2.1.B28 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.B28

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli	Pyrococcus furiosus

Protein Variants

Protein Variants	Comment	Organism
N206N	about 300fold decrease in catalytic efficiency for 4-nitrophenyl-beta-D-glucospyranoside, about 50fold decrease in catalytic efficiency for 4-nitrophenyl-beta-D-galactoside	Pyrococcus furiosus
R77Q	2000fold decrease in catalytic efficiency for 4-nitrophenyl-beta-D-glucospyranoside, 175fold decrease in catalytic efficiency for 4-nitrophenyl-beta-D-galactoside. The mutant enzyme shows a biphasic behavior for the hydrolysis of 4-nitrophenyl beta-D-mannopyranoside with separate kinetic parameters above and below 1 mM 4-nitrophenyl beta-D-mannopyranoside	Pyrococcus furiosus
R77Q/N206D	about 600fold decrease in catalytic efficiency for 4-nitrophenyl-beta-D-glucospyranoside, about 430fold decrease in catalytic efficiency for 4-nitrophenyl-beta-D-galactoside. The mutant enzyme shows a biphasic behavior for the hydrolysis of 4-nitrophenyl beta-D-mannopyranoside with separate kinetic parameters above and below 1 mM 4-nitrophenyl beta-D-mannopyranoside	Pyrococcus furiosus

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 furiosus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	mutants enzymes R77Q and R77Q/N206D show a biphasic behavior for the hydrolysis of 4-nitrophenyl beta-D-mannopyranoside with separate kinetic parameters above and below 1 mM 4-nitrophenyl beta-D-mannopyranoside	Pyrococcus furiosus
0.1	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme R77Q/N206D, below 1 mM 4-nitrophenyl beta-D-mannopyranoside. The mutant enzyme shows a biphasic behavior for the hydrolysis of 4-nitrophenyl beta-D-mannopyranoside with separate kinetic parameters above and below 1 mM 4-nitrophenyl beta-D-mannopyranoside	Pyrococcus furiosus
0.15	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme R77Q, below 1 mM 4-nitrophenyl beta-D-mannopyranoside. The mutant enzyme shows a biphasic behavior for the hydrolysis of 4-nitrophenyl beta-D-mannopyranoside with separate kinetic parameters above and below 1 mM 4-nitrophenyl beta-D-mannopyranoside	Pyrococcus furiosus
0.19	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, wild-type enzyme	Pyrococcus furiosus
0.63	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme N206D	Pyrococcus furiosus
0.64	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme R77Q, above 1 mM 4-nitrophenyl beta-D-mannopyranoside. The mutant enzyme shows a biphasic behavior for the hydrolysis of 4-nitrophenyl beta-D-mannopyranoside with separate kinetic parameters above and below 1 mM 4-nitrophenyl beta-D-mannopyranoside	Pyrococcus furiosus
1.3	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, wild-type enzyme	Pyrococcus furiosus
1.5	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme R77Q/N206D, above 1 mM 4-nitrophenyl beta-D-mannopyranoside. The mutant enzyme shows a biphasic behavior for the hydrolysis of 4-nitrophenyl beta-D-mannopyranoside with separate kinetic parameters above and below 1 mM 4-nitrophenyl beta-D-mannopyranoside	Pyrococcus furiosus
4.6	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, mutant enzyme N206D	Pyrococcus furiosus
5	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, wild-type enzyme	Pyrococcus furiosus
13.8	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, mutant enzyme R77Q	Pyrococcus furiosus
13.9	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, mutant enzyme R77Q/N206D	Pyrococcus furiosus
15.7	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, mutant enzyme N206D	Pyrococcus furiosus
21.7	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, mutant enzyme R77Q	Pyrococcus furiosus
22	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, mutant enzyme R77Q/N206D	Pyrococcus furiosus

Organism

Organism	UniProt	Comment	Textmining
Pyrococcus furiosus	Q51723	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Pyrococcus furiosus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
4-nitrophenyl beta-D-galactopyranoside + H2O	catalytic efficiency (kcat/Km) for 4-nitrophenyl beta-D-galactopyranoside is 7.6% compared to catalytic catalytic efficiency (kcat/Km) for 4-nitrophenyl beta-D-glucopyranoside	Pyrococcus furiosus	4-nitrophenol + beta-D-galactose	-	?
4-nitrophenyl beta-D-glucopyranoside + H2O	-	Pyrococcus furiosus	4-nitrophenol + beta-D-glucose	-	?
4-nitrophenyl beta-D-mannopyranoside + H2O	catalytic efficiency (kcat/Km) for 4-nitrophenyl beta-D-mannopyranoside is 0.7% compared to catalytic catalytic efficiency (kcat/Km) for 4-nitrophenyl beta-D-glucopyranoside	Pyrococcus furiosus	4-nitrophenol + beta-D-mannose	-	?

Synonyms

Synonyms	Comment	Organism
CelB	-	Pyrococcus furiosus

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
102	-	half-life: over 10 h	Pyrococcus furiosus
106	-	melting temperature	Pyrococcus furiosus

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
additional information	-	additional information	mutants enzymes R77Q and R77Q/N206D show a biphasic behavior for the hydrolysis of 4-nitrophenyl beta-D-mannopyranoside with separate kinetic parameters above and below 1 mM 4-nitrophenyl beta-D-mannopyranoside	Pyrococcus furiosus
0.85	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme R77Q, below 1 mM 4-nitrophenyl beta-D-mannopyranoside. The mutant enzyme shows a biphasic behavior for the hydrolysis of 4-nitrophenyl beta-D-mannopyranoside with separate kinetic parameters above and below 1 mM 4-nitrophenyl beta-D-mannopyranoside	Pyrococcus furiosus
1.02	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme R77Q/N206D, below 1 mM 4-nitrophenyl beta-D-mannopyranoside. The mutant enzyme shows a biphasic behavior for the hydrolysis of 4-nitrophenyl beta-D-mannopyranoside with separate kinetic parameters above and below 1 mM 4-nitrophenyl beta-D-mannopyranoside	Pyrococcus furiosus
1.22	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme R77Q, above 1 mM 4-nitrophenyl beta-D-mannopyranoside. The mutant enzyme shows a biphasic behavior for the hydrolysis of 4-nitrophenyl beta-D-mannopyranoside with separate kinetic parameters above and below 1 mM 4-nitrophenyl beta-D-mannopyranoside	Pyrococcus furiosus
1.9	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme R77Q/N206D, above 1 mM 4-nitrophenyl beta-D-mannopyranoside. The mutant enzyme shows a biphasic behavior for the hydrolysis of 4-nitrophenyl beta-D-mannopyranoside with separate kinetic parameters above and below 1 mM 4-nitrophenyl beta-D-mannopyranoside	Pyrococcus furiosus
1.97	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme N206D	Pyrococcus furiosus
29.2	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, mutant enzyme R77Q/N206D	Pyrococcus furiosus
65.9	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, wild-type enzyme	Pyrococcus furiosus
69.2	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, mutant enzyme R77Q	Pyrococcus furiosus
116	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, mutant enzyme N206D	Pyrococcus furiosus
172	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, mutant enzyme R77Q/N206D	Pyrococcus furiosus
180	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, mutant enzyme N206D	Pyrococcus furiosus
207	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, mutant enzyme R77Q	Pyrococcus furiosus
1140	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, wild-type enzyme	Pyrococcus furiosus
2827	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, wild-type enzyme	Pyrococcus furiosus

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.0000065	-	(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 furiosus
0.000097	-	(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 R77Q	Pyrococcus furiosus
0.0027	-	(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 R77Q/N206D	Pyrococcus furiosus
0.0098	-	(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 N206D	Pyrococcus furiosus

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
additional information	-	additional information	mutants enzymes R77Q and R77Q/N206D show a biphasic behavior for the hydrolysis of 4-nitrophenyl beta-D-mannopyranoside with separate kinetic parameters above and below 1 mM 4-nitrophenyl beta-D-mannopyranoside	Pyrococcus furiosus
1.3	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, mutant enzyme R77Q/N206D	Pyrococcus furiosus
1.3	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme R77Q/N206D, above 1 mM 4-nitrophenyl beta-D-mannopyranoside. The mutant enzyme shows a biphasic behavior for the hydrolysis of 4-nitrophenyl beta-D-mannopyranoside with separate kinetic parameters above and below 1 mM 4-nitrophenyl beta-D-mannopyranoside	Pyrococcus furiosus
1.9	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme R77Q, above 1 mM 4-nitrophenyl beta-D-mannopyranoside. The mutant enzyme shows a biphasic behavior for the hydrolysis of 4-nitrophenyl beta-D-mannopyranoside with separate kinetic parameters above and below 1 mM 4-nitrophenyl beta-D-mannopyranoside	Pyrococcus furiosus
3.1	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme N206D	Pyrococcus furiosus
3.2	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, mutant enzyme R77Q	Pyrococcus furiosus
5.7	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme R77Q, below 1 mM 4-nitrophenyl beta-D-mannopyranoside. The mutant enzyme shows a biphasic behavior for the hydrolysis of 4-nitrophenyl beta-D-mannopyranoside with separate kinetic parameters above and below 1 mM 4-nitrophenyl beta-D-mannopyranoside	Pyrococcus furiosus
10.2	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, mutant enzyme R77Q/N206D, below 1 mM 4-nitrophenyl beta-D-mannopyranoside. The mutant enzyme shows a biphasic behavior for the hydrolysis of 4-nitrophenyl beta-D-mannopyranoside with separate kinetic parameters above and below 1 mM 4-nitrophenyl beta-D-mannopyranoside	Pyrococcus furiosus
11.5	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, mutant enzyme N206D	Pyrococcus furiosus
12.3	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, mutant enzyme R77Q/N206D	Pyrococcus furiosus
14.9	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, mutant enzyme R77Q	Pyrococcus furiosus
24.9	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, mutant enzyme N206D	Pyrococcus furiosus
49.8	-	4-nitrophenyl beta-D-mannopyranoside	pH 5.0, 90°C, wild-type enzyme	Pyrococcus furiosus
561	-	4-nitrophenyl beta-D-galactopyranoside	pH 5.0, 90°C, wild-type enzyme	Pyrococcus furiosus
7337	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.0, 90°C, wild-type enzyme	Pyrococcus furiosus

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Release 2023.1 (January 2023)