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Literature summary for 3.2.1.4 extracted from

  • Kang, H.J.; Ishikawa, K.
    Analysis of active center in hyperthermophilic cellulase from Pyrococcus horikoshii (2007), J. Microbiol. Biotechnol., 17, 1249-1253.
    View publication on PubMed

Protein Variants

Protein Variants Comment Organism
D385N activity towards carboxymethyl cellulose is 29.9% of wild-type activity Pyrococcus horikoshii
DELTAQ1-G5 activity towards carboxymethyl cellulose is 135.6% of wild-type activity. kcat/Km for p-nitrophenyl cellobiose is 2.3fold higher than wild-type value. Thermostability is not significantly influenced Pyrococcus horikoshii
E201A activity towards carboxymethyl cellulose is 0.01% of wild-type activity Pyrococcus horikoshii
E201Q activity towards carboxymethyl cellulose is 1.12% of wild-type activity. kcat/Km for p-nitrophenyl cellobiose is 43fold lower than wild-type value Pyrococcus horikoshii
E342A activity towards carboxymethyl cellulose is 0.08% of wild-type activity, no activity with p-nitrophenyl cellobiose Pyrococcus horikoshii
E342Q activity towards carboxymethyl cellulose is 0.01% of wild-type activity Pyrococcus horikoshii
H297A activity towards carboxymethyl cellulose is 0.08% of wild-type activity Pyrococcus horikoshii
H297N activity towards carboxymethyl cellulose is 1.31% of wild-type activity. pH-optimum is 7.0, compared to 5.5-6 for wild-type enzyme Pyrococcus horikoshii
additional information mutant enzyme lacking 5 residues at the C-terminus: hydrolytic activity towards carboxymethyl cellulose is 112% of wild-type activity, kcat/Km for p-nitrophenyl cellobiose is 1.2fold higher than wild-type value. Mutant enzyme lacking 5 residues at the C-terminus and 5 residues at the N-terminus: activity towards carboxymethyl cellulose is 111% of wild-type activity, kcat/Km for p-nitrophenyl cellobiose is 1.8fold higher than wild-type value. Thermostability is not significantly influenced Pyrococcus horikoshii
N200A activity towards carboxymethyl cellulose is 5.43% of wild-type activity Pyrococcus horikoshii
R102A activity towards carboxymethyl cellulose is 0.67% of wild-type activity Pyrococcus horikoshii
W377A activity towards carboxymethyl cellulose is 1.02% of wild-type activity Pyrococcus horikoshii
Y299A activity towards carboxymethyl cellulose is 0.21% of wild-type activity Pyrococcus horikoshii
Y299F activity towards carboxymethyl cellulose is 2.15% of wild-type activity. pH-optimum is 8.5, compared to 5.5-6 for wild-type enzyme Pyrococcus horikoshii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.76
-
p-nitrophenyl cellobiose pH 5-6, 50°C, mutant enzyme DELTAQ1-G5 Pyrococcus horikoshii
0.95
-
p-nitrophenyl cellobiose pH 5-6, 50°C, wild-type enzyme Pyrococcus horikoshii
1.12
-
p-nitrophenyl cellobiose pH 5-6, 50°C, mutant enzyme Y299F Pyrococcus horikoshii
1.13
-
p-nitrophenyl cellobiose pH 5-6, 50°C, mutant enzyme lacking 5 residues at the C-terminus and 5 residies at the N-terminus Pyrococcus horikoshii
1.24
-
p-nitrophenyl cellobiose pH 5-6, 50°C, mutant enzyme E201Q Pyrococcus horikoshii
1.44
-
p-nitrophenyl cellobiose pH 5-6, 50°C, mutant enzyme lacking 5 residues at the C-terminus Pyrococcus horikoshii
2.4
-
p-nitrophenyl cellobiose pH 5-6, 50°C, mutant enzyme D385N Pyrococcus horikoshii
2.76
-
p-nitrophenyl cellobiose pH 5-6, 50°C, mutant enzyme H297N Pyrococcus horikoshii

Organism

Organism UniProt Comment Textmining
Pyrococcus horikoshii O58925
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
carboxymethyl cellulose + H2O
-
Pyrococcus horikoshii ?
-
?
p-nitrophenyl cellobiose + H2O
-
Pyrococcus horikoshii p-nitrophenol + cellobiose
-
?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.01
-
p-nitrophenyl cellobiose pH 5-6, 50°C, mutant enzyme E201Q Pyrococcus horikoshii
0.01
-
p-nitrophenyl cellobiose pH 5-6, 50°C, mutant enzyme H297N Pyrococcus horikoshii
0.01
-
p-nitrophenyl cellobiose pH 5-6, 50°C, mutant enzyme Y299F Pyrococcus horikoshii
0.11
-
p-nitrophenyl cellobiose pH 5-6, 50°C, mutant enzyme D385N Pyrococcus horikoshii
0.41
-
p-nitrophenyl cellobiose pH 5-6, 50°C, wild-type enzyme Pyrococcus horikoshii
0.75
-
p-nitrophenyl cellobiose pH 5-6, 50°C, mutant enzyme lacking 5 residues at the C-terminus Pyrococcus horikoshii
0.78
-
p-nitrophenyl cellobiose pH 5-6, 50°C, mutant enzyme DELTAQ1-G5 Pyrococcus horikoshii
0.91
-
p-nitrophenyl cellobiose pH 5-6, 50°C, mutant enzyme lacking 5 residues at the C-terminus and 5 residies at the N-terminus Pyrococcus horikoshii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.5 6 optimal pH for wild-type enzyme and mutant enzymes E201Q, E342Q, and D385N Pyrococcus horikoshii
7
-
mutant enzyme H297N Pyrococcus horikoshii
8.5
-
mutant enzyme Y299F Pyrococcus horikoshii