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Literature summary for 3.2.1.B28 extracted from

  • Perugino, G.; Trincone, A.; Giordano, A.; van der Oost, J.; Kaper, T.; Rossi, M.; Moracci, M.
    Activity of hyperthermophilic glycosynthases is significantly enhanced at acidic pH (2003), Biochemistry, 42, 8484-8493.
    View publication on PubMed
Search for more literature for 3.2.1.B28

Protein Variants

Protein Variants Comment Organism
E386G mutation in nucleophile residue E387, mutation completely abolishes activity under statndard conditions. The addition of 2 M sodium formate as an external nucleophile leads to the recovery of 8.40% activity with accumulation of oligosaccharides. At pH 3.0 and low concentrations of sodium formate buffer, the hyperthermophilic glycosynthase shows kcat values similar to those of the wild-type and 17fold higher than those observed at the usual reactivation conditions in 2 M sodium formate at pH 6.5 Pyrococcus furiosus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.22
-
 2-nitrophenyl beta-D-glucopyranoside wild-type, pH 6.5, 65°C, presence of 2 M formate Pyrococcus furiosus
0.28
-
 2-nitrophenyl beta-D-glucopyranoside wild-type, pH 6.5, 65°C Pyrococcus furiosus
1.3
-
 2-nitrophenyl beta-D-glucopyranoside mutant E372A, pH 5.0, 65°C Pyrococcus furiosus
1.49
-
 2-nitrophenyl beta-D-glucopyranoside mutant E372A, pH 6.5, 65°C, presence of 2 M formate Pyrococcus furiosus
2.1
-
 2-nitrophenyl beta-D-glucopyranoside mutant E372A, pH 4.0, 65°C Pyrococcus furiosus
4.3
-
 2-nitrophenyl beta-D-glucopyranoside mutant E372A, pH 3.0, 65°C Pyrococcus furiosus

Organism

Organism UniProt Comment Textmining
Pyrococcus furiosus Q51723
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.08
-
 mutant E372A, substrate 2-nitrophenyl beta-D-galactopyranoside, pH 3.0, 65°C Pyrococcus furiosus
0.28
-
 mutant E372A, substrate 2-nitrophenyl beta-D-xylopyranoside, pH 3.0, 65°C Pyrococcus furiosus
4.7
-
 mutant E372A, substrate 2-nitrophenyl beta-D-glucopyranoside, pH 3.0, 65°C Pyrococcus furiosus
8.3
-
 mutant E372A, substrate 2,4-dinitrophenyl beta-D-glucopyranoside, pH 3.0, 65°C Pyrococcus furiosus
66
-
 wild-type, substrate cellobiose, pH 3.0, 65°C Pyrococcus furiosus
75
-
 wild-type, substrate 2-nitrophenyl beta-D-xylopyranoside, pH 3.0, 65°C Pyrococcus furiosus
291
-
 wild-type, substrate 4-nitrophenyl beta-D-glucopyranoside, pH 3.0, 65°C Pyrococcus furiosus
320
-
 wild-type, substrate 2,4-dinitrophenyl beta-D-glucopyranoside, pH 3.0, 65°C Pyrococcus furiosus
534
-
 wild-type, substrate 2-nitrophenyl beta-D-glucopyranoside, pH 3.0, 65°C Pyrococcus furiosus
1049
-
 wild-type, substrate 2-nitrophenyl beta-D-galactopyranoside, pH 3.0, 65°C Pyrococcus furiosus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2,4-dinitrophenyl beta-D-glucopyranoside + H2O
-
 Pyrococcus furiosus 2,4-dinitrophenol + D-glucopyranose
-
 ?
2-nitrophenyl beta-D-galactopyranoside + H2O
-
 Pyrococcus furiosus 2-nitrophenol + D-galactopyranose
-
 ?
2-nitrophenyl beta-D-glucopyranoside + H2O
-
 Pyrococcus furiosus 2-nitrophenol + D-glucopyranose
-
 ?
2-nitrophenyl beta-D-xylopyranoside + H2O
-
 Pyrococcus furiosus 2-nitrophenol + D-xylopyranose
-
 ?
4-nitrophenyl beta-D-glucopyranoside + H2O
-
 Pyrococcus furiosus 4-nitrophenol + D-glucopyranose
-
 ?
cellobiose + H2O
-
 Pyrococcus furiosus 2 D-glucose
-
 ?

Synonyms

Synonyms Comment Organism
CelB
-
 Pyrococcus furiosus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.45
-
 2-nitrophenyl beta-D-glucopyranoside mutant E372A, pH 5.0, 65°C Pyrococcus furiosus
3.9
-
 2-nitrophenyl beta-D-glucopyranoside mutant E372A, pH 6.5, 65°C, presence of 2 M formate Pyrococcus furiosus
6.7
-
 2-nitrophenyl beta-D-glucopyranoside mutant E372A, pH 4.0, 65°C Pyrococcus furiosus
47.6
-
 2-nitrophenyl beta-D-glucopyranoside mutant E372A, pH 3.0, 65°C Pyrococcus furiosus
1189
-
 2-nitrophenyl beta-D-glucopyranoside wild-type, pH 6.5, 65°C, presence of 2 M formate Pyrococcus furiosus
1797
-
 2-nitrophenyl beta-D-glucopyranoside wild-type, pH 6.5, 65°C Pyrococcus furiosus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.3
-
 2-nitrophenyl beta-D-glucopyranoside mutant E372A, pH 5.0, 65°C Pyrococcus furiosus
2.6
-
 2-nitrophenyl beta-D-glucopyranoside mutant E372A, pH 6.5, 65°C, presence of 2 M formate Pyrococcus furiosus
3.1
-
 2-nitrophenyl beta-D-glucopyranoside mutant E372A, pH 4.0, 65°C Pyrococcus furiosus
10.9
-
 2-nitrophenyl beta-D-glucopyranoside mutant E372A, pH 3.0, 65°C Pyrococcus furiosus
5350
-
 2-nitrophenyl beta-D-glucopyranoside wild-type, pH 6.5, 65°C, presence of 2 M formate Pyrococcus furiosus
6480
-
 2-nitrophenyl beta-D-glucopyranoside wild-type, pH 6.5, 65°C Pyrococcus furiosus