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Literature summary for 3.2.1.B28 extracted from

  • Kaper, T.; Lebbink, J.H.G.; Pouwels, J.; Kopp, J.; Schulz, G.E.; van der Oost, J.; de Vos, W.M.
    Comparative structural analysis and substrate specificity engineering of the hyperthermostable beta-glucosidase CelB from Pyrococcus furiosus (2000), Biochemistry, 39, 4963-4970.
    View publication on PubMed
Search for more literature for 3.2.1.B28

Crystallization (Commentary)

Crystallization (Comment) Organism
3.3 A resolution structural model. CelB shows a homo-tetramer configuration, with subunits having a typical (betaalpha)8-barrel fold Pyrococcus furiosus

Protein Variants

Protein Variants Comment Organism
E417S mutation in phosphate binding site, 5fold increase of the efficiency of hydrolyzing o-nitrophenol-beta-D-galactopyranoside-6-phosphate. Activity on nonphosphorylated sugars is largely reduced Pyrococcus furiosus
E417S/M424K/F426Y mutations in phosphate binding site, 3fold increase of the efficiency of hydrolyzing 2-nitrophenyl beta-D-galactopyranoside-6-phosphate. Activity on nonphosphorylated sugars is largely reduced Pyrococcus furiosus
F426Y mutation in phosphate binding site, results in an increased affinity for galactosides. Activity on nonphosphorylated sugars is largely reduced Pyrococcus furiosus
M424K mutation in phosphate binding site, shifting pH optimum from 5.0 to 6.0. Activity on nonphosphorylated sugars is largely reduced Pyrococcus furiosus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.28
-
 2-nitrophenyl beta-D-glucopyranoside wild-type, pH 5.0, 90°C Pyrococcus furiosus
0.51
-
 2-nitrophenyl beta-D-glucopyranoside mutant M424K, pH 5.0, 90°C Pyrococcus furiosus
0.57
-
 2-nitrophenyl beta-D-glucopyranoside mutant F426Y, pH 5.0, 90°C Pyrococcus furiosus
0.89
-
 2-nitrophenyl beta-D-galactopyranoside mutant F426Y, pH 5.0, 90°C Pyrococcus furiosus
2.3
-
 2-nitrophenyl beta-D-galactopyranoside wild-type, pH 5.0, 90°C Pyrococcus furiosus
4.8
-
 2-nitrophenyl beta-D-galactopyranoside mutant M424K, pH 5.0, 90°C Pyrococcus furiosus
16
-
 2-nitrophenyl beta-D-galactopyranoside-6-phosphate mutant E417S/M424K/F426Y, pH 5.0, 90°C Pyrococcus furiosus
22
-
 2-nitrophenyl beta-D-galactopyranoside-6-phosphate mutant E417S, pH 5.0, 90°C Pyrococcus furiosus
29
-
 2-nitrophenyl beta-D-glucopyranoside mutant E417S, pH 5.0, 90°C Pyrococcus furiosus
31
-
 2-nitrophenyl beta-D-galactopyranoside-6-phosphate wild-type, pH 5.0, 90°C Pyrococcus furiosus
38
-
 2-nitrophenyl beta-D-galactopyranoside-6-phosphate mutant F426Y, pH 5.0, 90°C Pyrococcus furiosus
40
-
 2-nitrophenyl beta-D-galactopyranoside mutant E417S/M424K/F426Y, pH 5.0, 90°C Pyrococcus furiosus
42
-
 2-nitrophenyl beta-D-galactopyranoside mutant E417S, pH 5.0, 90°C Pyrococcus furiosus
59
-
 2-nitrophenyl beta-D-glucopyranoside mutant E417S/M424K/F426Y, pH 5.0, 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.
2-nitrophenyl beta-D-galactopyranoside + H2O
-
 Pyrococcus furiosus 2-nitrophenol + D-galactopyranoside
-
 ?
2-nitrophenyl beta-D-galactopyranoside-6-phosphate + H2O
-
 Pyrococcus furiosus 2-nitrophenol + D-galactopyranoside-6-phosphate
-
 ?
2-nitrophenyl beta-D-glucopyranoside + H2O
-
 Pyrococcus furiosus 2-nitrophenol + D-glucopyranoside
-
 ?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
20 50 2-nitrophenyl beta-D-galactopyranoside wild-type, pH 5.0, 90°C Pyrococcus furiosus
26
-
 2-nitrophenyl beta-D-galactopyranoside-6-phosphate mutant F426Y, pH 5.0, 90°C Pyrococcus furiosus
49
-
 2-nitrophenyl beta-D-galactopyranoside-6-phosphate wild-type, pH 5.0, 90°C Pyrococcus furiosus
69
-
 2-nitrophenyl beta-D-galactopyranoside-6-phosphate mutant E417S/M424K/F426Y, pH 5.0, 90°C Pyrococcus furiosus
171
-
 2-nitrophenyl beta-D-galactopyranoside-6-phosphate mutant E417S, pH 5.0, 90°C Pyrococcus furiosus
175
-
 2-nitrophenyl beta-D-galactopyranoside mutant E417S/M424K/F426Y, pH 5.0, 90°C Pyrococcus furiosus
772
-
 2-nitrophenyl beta-D-glucopyranoside mutant M424K, pH 5.0, 90°C Pyrococcus furiosus
777
-
 2-nitrophenyl beta-D-glucopyranoside mutant E417S/M424K/F426Y, pH 5.0, 90°C Pyrococcus furiosus
840
-
 2-nitrophenyl beta-D-glucopyranoside mutant F426Y, pH 5.0, 90°C Pyrococcus furiosus
860
-
 2-nitrophenyl beta-D-glucopyranoside wild-type, pH 5.0, 90°C Pyrococcus furiosus
892
-
 2-nitrophenyl beta-D-galactopyranoside mutant F426Y, pH 5.0, 90°C Pyrococcus furiosus
1060
-
 2-nitrophenyl beta-D-galactopyranoside mutant E417S, pH 5.0, 90°C Pyrococcus furiosus
1880
-
 2-nitrophenyl beta-D-galactopyranoside mutant M424K, pH 5.0, 90°C Pyrococcus furiosus
3300
-
 2-nitrophenyl beta-D-glucopyranoside mutant E417S, pH 5.0, 90°C Pyrococcus furiosus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
4
-
 wild-type, substrate 2-nitrophenyl beta-D-galactopyranoside-6-phosphate Pyrococcus furiosus
5
-
 mutant E417S/M424K/F426Y, substrates 2-nitrophenyl beta-D-glucopyranoside, 2-nitrophenyl beta-D-galactopyranoside Pyrococcus furiosus
5
-
 wild-type, substrates 2-nitrophenyl beta-D-glucopyranoside, 2-nitrophenyl beta-D-galactopyranoside Pyrococcus furiosus
6
-
 mutant E417S/M424K/F426Y, substrate 2-nitrophenyl beta-D-galactopyranoside-6-phosphate Pyrococcus furiosus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.7
-
 2-nitrophenyl beta-D-galactopyranoside-6-phosphate mutant F426Y, pH 5.0, 90°C Pyrococcus furiosus
1.6
-
 2-nitrophenyl beta-D-galactopyranoside-6-phosphate wild-type, pH 5.0, 90°C Pyrococcus furiosus
4.2
-
 2-nitrophenyl beta-D-galactopyranoside-6-phosphate mutant E417S/M424K/F426Y, pH 5.0, 90°C Pyrococcus furiosus
4.4
-
 2-nitrophenyl beta-D-galactopyranoside mutant E417S/M424K/F426Y, pH 5.0, 90°C Pyrococcus furiosus
7.7
-
 2-nitrophenyl beta-D-galactopyranoside-6-phosphate mutant E417S, pH 5.0, 90°C Pyrococcus furiosus
13
-
 2-nitrophenyl beta-D-glucopyranoside mutant E417S/M424K/F426Y, pH 5.0, 90°C Pyrococcus furiosus
25.3
-
 2-nitrophenyl beta-D-galactopyranoside mutant E417S, pH 5.0, 90°C Pyrococcus furiosus
113
-
 2-nitrophenyl beta-D-glucopyranoside mutant E417S, pH 5.0, 90°C Pyrococcus furiosus
396
-
 2-nitrophenyl beta-D-galactopyranoside mutant M424K, pH 5.0, 90°C Pyrococcus furiosus
899
-
 2-nitrophenyl beta-D-galactopyranoside wild-type, pH 5.0, 90°C Pyrococcus furiosus
1000
-
 2-nitrophenyl beta-D-galactopyranoside mutant F426Y, pH 5.0, 90°C Pyrococcus furiosus
1484
-
 2-nitrophenyl beta-D-glucopyranoside mutant F426Y, pH 5.0, 90°C Pyrococcus furiosus
1510
-
 2-nitrophenyl beta-D-glucopyranoside mutant M424K, pH 5.0, 90°C Pyrococcus furiosus
4048
-
 2-nitrophenyl beta-D-glucopyranoside wild-type, pH 5.0, 90°C Pyrococcus furiosus