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Literature summary for 3.2.1.B28 extracted from

  • Lebbink, J.H.; Kaper, T.; Bron, P.; van der Oost, J.; de Vos, W.M.
    Improving low-temperature catalysis in the hyperthermostable Pyrococcus furiosus beta-glucosidase CelB by directed evolution (2000), Biochemistry, 39, 3656-3665.
    View publication on PubMed
Search for more literature for 3.2.1.B28

Protein Variants

Protein Variants Comment Organism
A419T 159% of wild-type activity with 4-nitrophenyl beta-D-glucoside, 109% with 4-nitrophenyl beta-D-galactoside, 53% residual activity after 1 h at 106°C Pyrococcus furiosus
I161V 110% of wild-type activity with 4-nitrophenyl beta-D-glucoside, 37% with 4-nitrophenyl beta-D-galactoside, 84% residual activity after 1 h at 106°C Pyrococcus furiosus
I67T 110% of wild-type activity with 4-nitrophenyl beta-D-glucoside, 97% with 4-nitrophenyl beta-D-galactoside, 100% residual activity after 1 h at 106°C Pyrococcus furiosus
K285R 155% of wild-type activity with 4-nitrophenyl beta-D-glucoside, 103% with 4-nitrophenyl beta-D-galactoside, 58% residual activity after 1 h at 106°C Pyrococcus furiosus
K70R 175% of wild-type activity with 4-nitrophenyl beta-D-glucoside, 147% with 4-nitrophenyl beta-D-galactoside, 1% residual activity after 1 h at 106°C Pyrococcus furiosus
M424V 199% of wild-type activity with 4-nitrophenyl beta-D-glucoside, 140% with 4-nitrophenyl beta-D-galactoside, 24% residual activity after 1 h at 106°C Pyrococcus furiosus
N415S 259% of wild-type activity with 4-nitrophenyl beta-D-glucoside, 64% with 4-nitrophenyl beta-D-galactoside. 7.5fold increase in the ratio of 4-nitrophenyl beta-D-glucopyranoside/p-nitrophenyl beta-D-galactopyranoside hydrolysis, no decrease in thermostability Pyrococcus furiosus
T371A 148% of wild-type activity with 4-nitrophenyl beta-D-glucoside, 149% with 4-nitrophenyl beta-D-galactoside, 14% residual activity after 1 h at 106°C Pyrococcus furiosus
V211A 260% of wild-type activity with 4-nitrophenyl beta-D-glucoside, 151% with 4-nitrophenyl beta-D-galactoside, 1% residual activity after 1 h at 106°C Pyrococcus furiosus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.42
-
 4-nitrophenyl beta-D-glucopyranoside wild-type, pH 4.8, 90°C Pyrococcus furiosus
0.53
-
 4-nitrophenyl beta-D-glucopyranoside wild-type, pH 4.8, 20°C Pyrococcus furiosus
0.57
-
 4-nitrophenyl beta-D-glucopyranoside mutant T371A, pH 4.8, 20°C Pyrococcus furiosus
0.7
-
 4-nitrophenyl beta-D-galactopyranoside wild-type, pH 4.8, 20°C Pyrococcus furiosus
0.92
-
 4-nitrophenyl beta-D-glucopyranoside mutant M424V, pH 4.8, 20°C Pyrococcus furiosus
1.1
-
 4-nitrophenyl beta-D-glucopyranoside mutant N4515S, pH 4.8, 20°C Pyrococcus furiosus
1.2
-
 4-nitrophenyl beta-D-galactopyranoside mutant N4515S, pH 4.8, 20°C Pyrococcus furiosus
1.3
-
 4-nitrophenyl beta-D-glucopyranoside mutant A419T, pH 4.8, 20°C Pyrococcus furiosus
1.3
-
 4-nitrophenyl beta-D-galactopyranoside mutant T371A, pH 4.8, 20°C Pyrococcus furiosus
1.4
-
 4-nitrophenyl beta-D-glucopyranoside mutant N4515S, pH 4.8, 90°C Pyrococcus furiosus
2 3 cellobiose wild-type, pH 4.8, 20°C Pyrococcus furiosus
3.4
-
 4-nitrophenyl beta-D-galactopyranoside mutant M424V, pH 4.8, 20°C Pyrococcus furiosus
4.5
-
 4-nitrophenyl beta-D-galactopyranoside mutant A419T, pH 4.8, 20°C Pyrococcus furiosus
5
-
 4-nitrophenyl beta-D-galactopyranoside wild-type, pH 4.8, 90°C Pyrococcus furiosus
8.5
-
 4-nitrophenyl beta-D-galactopyranoside mutant N4515S, pH 4.8, 90°C Pyrococcus furiosus
11
-
 cellobiose mutant T371A, pH 4.8, 20°C Pyrococcus furiosus
14
-
 cellobiose wild-type, pH 4.8, 90°C Pyrococcus furiosus
16
-
 cellobiose mutant M424V, pH 4.8, 20°C Pyrococcus furiosus
20
-
 cellobiose mutant N4515S, pH 4.8, 20°C Pyrococcus furiosus
25
-
 cellobiose mutant A419T, pH 4.8, 20°C Pyrococcus furiosus
38
-
 cellobiose mutant N4515S, pH 4.8, 90°C Pyrococcus furiosus
52
-
 lactose mutant T371A, pH 4.8, 20°C Pyrococcus furiosus
52
-
 lactose wild-type, pH 4.8, 20°C Pyrococcus furiosus
83
-
 lactose mutant A419T, pH 4.8, 20°C Pyrococcus furiosus
95
-
 lactose mutant N4515S, pH 4.8, 20°C Pyrococcus furiosus
97
-
 lactose mutant M424V, pH 4.8, 20°C Pyrococcus furiosus
120
-
 lactose wild-type, pH 4.8, 90°C Pyrococcus furiosus
220
-
 lactose mutant N4515S, pH 4.8, 90°C Pyrococcus furiosus

Organism

Organism UniProt Comment Textmining
Pyrococcus furiosus Q51723
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl beta-D-galactopyranoside + H2O
-
 Pyrococcus furiosus 4-nitrophenol + D-galactopyranose
-
 ?
4-nitrophenyl beta-D-glucopyranoside + H2O
-
 Pyrococcus furiosus 4-nitrophenol + D-glucopyranose
-
 ?
cellobiose + H2O
-
 Pyrococcus furiosus 2 D-glucose
-
 ?
lactose + H2O
-
 Pyrococcus furiosus D-glucose + D-galactose
-
 ?

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
98
-
-
 Pyrococcus furiosus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
106
-
 wild-type, 1 h, no loss of activity Pyrococcus furiosus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.2
-
 4-nitrophenyl beta-D-galactopyranoside mutant N4515S, pH 4.8, 20°C Pyrococcus furiosus
9.1
-
 4-nitrophenyl beta-D-galactopyranoside wild-type, pH 4.8, 20°C Pyrococcus furiosus
11
-
 4-nitrophenyl beta-D-galactopyranoside mutant T371A, pH 4.8, 20°C Pyrococcus furiosus
12
-
 4-nitrophenyl beta-D-galactopyranoside mutant M424V, pH 4.8, 20°C Pyrococcus furiosus
13
-
 lactose wild-type, pH 4.8, 20°C Pyrococcus furiosus
13
-
 4-nitrophenyl beta-D-glucopyranoside wild-type, pH 4.8, 20°C Pyrococcus furiosus
14
-
 4-nitrophenyl beta-D-galactopyranoside mutant A419T, pH 4.8, 20°C Pyrococcus furiosus
15
-
 lactose mutant A419T, pH 4.8, 20°C Pyrococcus furiosus
15
-
 cellobiose mutant T371A, pH 4.8, 20°C Pyrococcus furiosus
16
-
 cellobiose wild-type, pH 4.8, 20°C Pyrococcus furiosus
17
-
 lactose mutant T371A, pH 4.8, 20°C Pyrococcus furiosus
18
-
 cellobiose mutant M424V, pH 4.8, 20°C Pyrococcus furiosus
19
-
 cellobiose mutant A419T, pH 4.8, 20°C Pyrococcus furiosus
20
-
 cellobiose mutant N4515S, pH 4.8, 20°C Pyrococcus furiosus
21
-
 lactose mutant M424V, pH 4.8, 20°C Pyrococcus furiosus
21
-
 lactose mutant N4515S, pH 4.8, 20°C Pyrococcus furiosus
26
-
 4-nitrophenyl beta-D-glucopyranoside mutant T371A, pH 4.8, 20°C Pyrococcus furiosus
40
-
 4-nitrophenyl beta-D-glucopyranoside mutant A419T, pH 4.8, 20°C Pyrococcus furiosus
40
-
 4-nitrophenyl beta-D-glucopyranoside mutant M424V, pH 4.8, 20°C Pyrococcus furiosus
47
-
 4-nitrophenyl beta-D-glucopyranoside mutant N4515S, pH 4.8, 20°C Pyrococcus furiosus
240
-
 cellobiose mutant N4515S, pH 4.8, 90°C Pyrococcus furiosus
530
-
 lactose mutant N4515S, pH 4.8, 90°C Pyrococcus furiosus
670
-
 cellobiose wild-type, pH 4.8, 90°C Pyrococcus furiosus
1000
-
 4-nitrophenyl beta-D-galactopyranoside mutant N4515S, pH 4.8, 90°C Pyrococcus furiosus
1300
-
 lactose wild-type, pH 4.8, 90°C Pyrococcus furiosus
1400
-
 4-nitrophenyl beta-D-glucopyranoside mutant N4515S, pH 4.8, 90°C Pyrococcus furiosus
1600
-
 4-nitrophenyl beta-D-glucopyranoside wild-type, pH 4.8, 90°C Pyrococcus furiosus
2400
-
 4-nitrophenyl beta-D-galactopyranoside wild-type, pH 4.8, 90°C Pyrococcus furiosus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.18
-
 lactose mutant A419T, pH 4.8, 20°C Pyrococcus furiosus
0.22
-
 lactose mutant M424V, pH 4.8, 20°C Pyrococcus furiosus
0.22
-
 lactose mutant N4515S, pH 4.8, 20°C Pyrococcus furiosus
0.25
-
 lactose wild-type, pH 4.8, 20°C Pyrococcus furiosus
0.33
-
 lactose mutant T371A, pH 4.8, 20°C Pyrococcus furiosus
0.69
-
 cellobiose wild-type, pH 4.8, 20°C Pyrococcus furiosus
0.75
-
 cellobiose mutant A419T, pH 4.8, 20°C Pyrococcus furiosus
1
-
 cellobiose mutant N4515S, pH 4.8, 20°C Pyrococcus furiosus
1.1
-
 cellobiose mutant M424V, pH 4.8, 20°C Pyrococcus furiosus
1.4
-
 cellobiose mutant T371A, pH 4.8, 20°C Pyrococcus furiosus
2.4
-
 lactose mutant N4515S, pH 4.8, 90°C Pyrococcus furiosus
3.1
-
 4-nitrophenyl beta-D-galactopyranoside mutant A419T, pH 4.8, 20°C Pyrococcus furiosus
3.4
-
 4-nitrophenyl beta-D-galactopyranoside mutant M424V, pH 4.8, 20°C Pyrococcus furiosus
3.5
-
 4-nitrophenyl beta-D-galactopyranoside mutant N4515S, pH 4.8, 20°C Pyrococcus furiosus
8.2
-
 4-nitrophenyl beta-D-galactopyranoside mutant T371A, pH 4.8, 20°C Pyrococcus furiosus
11
-
 lactose wild-type, pH 4.8, 90°C Pyrococcus furiosus
13
-
 4-nitrophenyl beta-D-galactopyranoside wild-type, pH 4.8, 20°C Pyrococcus furiosus
25
-
 4-nitrophenyl beta-D-glucopyranoside wild-type, pH 4.8, 20°C Pyrococcus furiosus
31
-
 4-nitrophenyl beta-D-glucopyranoside mutant A419T, pH 4.8, 20°C Pyrococcus furiosus
43
-
 4-nitrophenyl beta-D-glucopyranoside mutant M424V, pH 4.8, 20°C Pyrococcus furiosus
43
-
 4-nitrophenyl beta-D-glucopyranoside mutant N4515S, pH 4.8, 20°C Pyrococcus furiosus
45
-
 4-nitrophenyl beta-D-glucopyranoside mutant T371A, pH 4.8, 20°C Pyrococcus furiosus
120
-
 4-nitrophenyl beta-D-galactopyranoside mutant N4515S, pH 4.8, 90°C Pyrococcus furiosus
480
-
 4-nitrophenyl beta-D-galactopyranoside wild-type, pH 4.8, 90°C Pyrococcus furiosus
1000
-
 cellobiose mutant N4515S, pH 4.8, 90°C Pyrococcus furiosus
1000
-
 4-nitrophenyl beta-D-glucopyranoside mutant N4515S, pH 4.8, 90°C Pyrococcus furiosus
2400
-
 cellobiose wild-type, pH 4.8, 90°C Pyrococcus furiosus
3900
-
 4-nitrophenyl beta-D-glucopyranoside wild-type, pH 4.8, 90°C Pyrococcus furiosus