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Literature summary for 3.2.1.14 extracted from

  • Tsuji, H.; Nishimura, S.; Inui, T.; Kado, Y.; Ishikawa, K.; Nakamura, T.; Uegaki, K.
    Kinetic and crystallographic analyses of the catalytic domain of chitinase from Pyrococcus furiosus - the role of conserved residues in the active site (2010), FEBS J., 277, 2683-2695.
    View publication on PubMed
Search for more literature for 3.2.1.14

Cloned(Commentary)

Cloned (Comment) Organism
overexpression of wild-type enzyme and mutant enzymes D522N, D522A, D524N, D524A, E526Q and E526A in Escherichia coli Pyrococcus furiosus

Crystallization (Commentary)

Crystallization (Comment) Organism
hanging drop vapor diffusion method, crystallographic analyses of the catalytic domain of chitinase. Crystals of these mutant enzymes E526A and D524A in complex with chito-oligosaccharide substrate by means of cocrystallization and soaking methods, respectively, and determination of their tertiary structures. X-ray diffraction data are collected, resolutions of 1.80 A and 1.76 A Pyrococcus furiosus

Protein Variants

Protein Variants Comment Organism
D522A mutation causes a 7.3fold decrease in kcat/Km for 4-nitrophenyl chitobioside Pyrococcus furiosus
D522N mutation causes a 3.4fold decrease in kcat/Km for 4-nitrophenyl chitobioside Pyrococcus furiosus
D524A mutation causes a 58.4 fold decrease in kcat/Km for 4-nitrophenyl chitobioside Pyrococcus furiosus
D524N mutation causes a 3.6fold decrease in kcat/Km for 4-nitrophenyl chitobioside Pyrococcus furiosus
E526A no activity detected with 4-nitrophenyl chitobioside Pyrococcus furiosus
E526Q mutation causes a 38.4fold decrease in kcat/Km for 4-nitrophenyl chitobioside Pyrococcus furiosus
W664A mutation causes a 8588fold decrease in kcat/Km for 4-nitrophenyl chitobioside Pyrococcus furiosus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.09
-
 4-nitrophenyl beta-N,N'-diacetylchitobioside pH 4.8, 50°C, mutant enzyme D524A Pyrococcus furiosus
0.12
-
 4-nitrophenyl beta-N,N'-diacetylchitobioside pH 4.8, 50°C, mutant enzyme E526Q Pyrococcus furiosus
0.46
-
 4-nitrophenyl beta-N,N'-diacetylchitobioside pH 4.8, 50°C, wild-type enzyme Pyrococcus furiosus
0.54
-
 4-nitrophenyl beta-N,N'-diacetylchitobioside pH 4.8, 50°C, mutant enzyme D522N Pyrococcus furiosus
0.61
-
 4-nitrophenyl beta-N,N'-diacetylchitobioside pH 4.8, 50°C, mutant enzyme D524N Pyrococcus furiosus
0.74
-
 4-nitrophenyl beta-N,N'-diacetylchitobioside pH 4.8, 50°C, mutant enzyme D522A Pyrococcus furiosus
12.7
-
 4-nitrophenyl beta-N,N'-diacetylchitobioside pH 4.8, 50°C, mutant enzyme W664A Pyrococcus furiosus

Organism

Organism UniProt Comment Textmining
Pyrococcus furiosus Q8U1H5
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Pyrococcus furiosus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl beta-N,N'-diacetylchitobioside + H2O
-
 Pyrococcus furiosus 4-nitrophenol + N,N'-diacetylchitobiose
-
 ?

Synonyms

Synonyms Comment Organism
PF1233 locus name Pyrococcus furiosus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
 assay at Pyrococcus furiosus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.022
-
 4-nitrophenyl beta-N,N'-diacetylchitobioside pH 4.8, 50°C, mutant enzyme D524A Pyrococcus furiosus
0.022
-
 4-nitrophenyl beta-N,N'-diacetylchitobioside pH 4.8, 50°C, mutant enzyme W664A Pyrococcus furiosus
0.045
-
 4-nitrophenyl beta-N,N'-diacetylchitobioside pH 4.8, 50°C, mutant enzyme E526Q Pyrococcus furiosus
1.49
-
 4-nitrophenyl beta-N,N'-diacetylchitobioside pH 4.8, 50°C, mutant enzyme D522A Pyrococcus furiosus
2.36
-
 4-nitrophenyl beta-N,N'-diacetylchitobioside pH 4.8, 50°C, mutant enzyme D522N Pyrococcus furiosus
2.47
-
 4-nitrophenyl beta-N,N'-diacetylchitobioside pH 4.8, 50°C, mutant enzyme D524N Pyrococcus furiosus
6.7
-
 4-nitrophenyl beta-N,N'-diacetylchitobioside pH 4.8, 50°C, wild-type enzyme Pyrococcus furiosus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
4.8
-
 assay at Pyrococcus furiosus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0017
-
 4-nitrophenyl chitobioside pH 4.8, 50°C, mutant enzyme W664A Pyrococcus furiosus
0.25
-
 4-nitrophenyl chitobioside pH 4.8, 50°C, mutant enzyme D524A Pyrococcus furiosus
0.38
-
 4-nitrophenyl chitobioside pH 4.8, 50°C, mutant enzyme E526Q Pyrococcus furiosus
2
-
 4-nitrophenyl chitobioside pH 4.8, 50°C, mutant enzyme D522A Pyrococcus furiosus
4.1
-
 4-nitrophenyl chitobioside pH 4.8, 50°C, mutant enzyme D524N Pyrococcus furiosus
4.3
-
 4-nitrophenyl chitobioside pH 4.8, 50°C, mutant enzyme D522N Pyrococcus furiosus
14.6
-
 4-nitrophenyl chitobioside pH 4.8, 50°C, wild-type enzyme Pyrococcus furiosus