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Literature summary for 1.1.99.18 extracted from

  • Rotsaert, F.A.J.; Renganathan, V.; Gold, M.H.
    Role of the flavin domain residues, His689 and Asn732, in the catalytic mechanism of cellobiose dehydrogenase from Phanerochaete chrysosporium (2003), Biochemistry, 42, 4049-4056.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
-
Phanerodontia chrysosporium

Protein Variants

Protein Variants Comment Organism
H689A more than 1000fold lower turnover value, Km-value for cellobiose and lactose is similar to that of the wild-type enzyme Phanerodontia chrysosporium
H689E more than 1000fold lower turnover value, Km-value for cellobiose and lactose is similar to that of the wild-type enzyme Phanerodontia chrysosporium
H689N more than 1000fold lower turnover value, Km-value for cellobiose and lactose is similar to that of the wild-type enzyme Phanerodontia chrysosporium
H689Q more than 1000fold lower turnover value, Km-value for cellobiose and lactose is similar to that of the wild-type enzyme Phanerodontia chrysosporium
H689V more than 1000fold lower turnover value, Km-value for cellobiose and lactose is similar to that of the wild-type enzyme Phanerodontia chrysosporium
N732A the turnover-number for cellobiose is 38.8fold lower than the turnover-number of the wild-type enzyme, the Km-value for cellobiose is 1.1fold higher than the KM-value of the wild-type enzyme, the turnover-number for lactose is 20.4fold lower than the turnover-number of the wild-type enzyme, the Km-value for lactose is 4.4fold higher than the KM-value of the wild-type enzyme Phanerodontia chrysosporium
N732D the turnover-number for cellobiose is 3875fold lower than the turnover-number of the wild-type enzyme, the Km-value for cellobiose is 10.6fold higher than the KM-value of the wild-type enzyme, the turnover-number for lactose is 2860fold lower than the turnover-number of the wild-type enzyme, the Km-value for lactose is 42.2fold higher than the KM-value of the wild-type enzyme. The pH optimum is shifted from pH 3-5 for the wild-type enzyme in the reaction with cellobiose and 2,6-dichlorophenol-indophenol to pH 6.5-7.0. The pH optimum is shifted from pH 3 for the wild-type enzyme in the reaction with cellobiose and cytochrome c to pH 6 Phanerodontia chrysosporium
N732E the turnover-number for cellobiose is 73.8fold lower than the turnover-number of the wild-type enzyme, the Km-value for cellobiose is 14.4fold higher than the KM-value of the wild-type enzyme, the turnover-number for lactose is 47.7fold lower than the turnover-number of the wild-type enzyme, the Km-value for lactose is 61.5fold higher than the KM-value of the wild-type enzyme. The pH optimum is shifted from pH 3-5 for the wild-type enzyme in the reaction with cellobiose and 2,6-dichlorophenol-indophenol to pH 6.5-7.0.The pH optimum is shifted from pH 3 for the wild-type enzyme in the reaction with cellobiose and cytochrome c to pH 5 Phanerodontia chrysosporium
N732H the turnover-number for cellobiose is 5.7fold lower than the turnover-number of the wild-type enzyme, the Km-value for cellobiose is 2.4fold higher than the KM-value of the wild-type enzyme, the turnover-number for lactose is 8.4fold lower than the turnover-number of the wild-type enzyme, the Km-value for lactose is 8.5fold higher than the KM-value of the wild-type enzyme Phanerodontia chrysosporium
N732Q the turnover-number for cellobiose is 15.5fold lower than the turnover-number of the wild-type enzyme, the Km-value for cellobiose is 2.6fold higher than the KM-value of the wild-type enzyme, the turnover-number for lactose is 11.9fold lower than the turnover-number of the wild-type enzyme, the Km-value for lactose is 14.8fold higher than the KM-value of the wild-type enzyme Phanerodontia chrysosporium

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.016
-
cellobiose pH 4.5, 23°C, recombinant wild-type enzyme Phanerodontia chrysosporium
0.017
-
cellobiose pH 4.5, 23°C, mutant enzyme N732A Phanerodontia chrysosporium
0.019
-
cellobiose pH 4.5, 23°C, mutant enzyme H689N Phanerodontia chrysosporium
0.021
-
cellobiose pH 4.5, 23°C, mutant enzyme H689Q or mutant enzyme H689A Phanerodontia chrysosporium
0.022
-
cellobiose pH 4.5, 23°C, mutant enzyme H689E Phanerodontia chrysosporium
0.031
-
cellobiose pH 4.5, 23°C, mutant enzyme H689V Phanerodontia chrysosporium
0.04
-
cellobiose pH 4.5, 23°C, mutant enzyme N732H Phanerodontia chrysosporium
0.041
-
cellobiose pH 4.5, 23°C, mutant enzyme N732Q Phanerodontia chrysosporium
0.17
-
cellobiose pH 4.5, 23°C, mutant enzyme N732D Phanerodontia chrysosporium
0.23
-
cellobiose pH 4.5, 23°C, mutant enzyme N732E Phanerodontia chrysosporium
0.27
-
lactose pH 4.5, 23°C, recombinant wild-type enzyme Phanerodontia chrysosporium
0.28
-
lactose pH 4.5, 23°C, mutant enzyme H689A Phanerodontia chrysosporium
0.29
-
lactose pH 4.5, 23°C, mutant enzyme H689Q Phanerodontia chrysosporium
1.2
-
lactose pH 4.5, 23°C, mutant enzyme N732A Phanerodontia chrysosporium
2.3
-
lactose pH 4.5, 23°C, mutant enzyme N732H Phanerodontia chrysosporium
4
-
lactose pH 4.5, 23°C, mutant enzyme N732Q Phanerodontia chrysosporium
11.4
-
lactose pH 4.5, 23°C, mutant enzyme N732D Phanerodontia chrysosporium
16.6
-
lactose pH 4.5, 23°C, mutant enzyme N732E Phanerodontia chrysosporium

Localization

Localization Comment Organism GeneOntology No. Textmining
extracellular
-
Phanerodontia chrysosporium
-
-

Organism

Organism UniProt Comment Textmining
Phanerodontia chrysosporium
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
cellobiose + 2,6-dichlorophenol-indophenol
-
Phanerodontia chrysosporium cellobiono-1,5-lactone + reduced 2,6-dichlorophenol-indophenol
-
?
cellobiose + cytochrome c
-
Phanerodontia chrysosporium cellobiono-1,5-lactone + reduced cytochrome c
-
?
lactose + cytochrome c lactose binds considerably more weakly than cellobiose Phanerodontia chrysosporium 4-O-(beta-D-galactopyranosyl)-D-glucono-1,5-lactone + reduced cytochrome c
-
?

Synonyms

Synonyms Comment Organism
Cdh
-
Phanerodontia chrysosporium
cellobiose dehydrogenase
-
Phanerodontia chrysosporium

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.002
-
cellobiose pH 4.5, 23°C, mutant enzyme H689V Phanerodontia chrysosporium
0.003
-
cellobiose pH 4.5, 23°C, mutant enzyme H689A Phanerodontia chrysosporium
0.003
-
lactose pH 4.5, 23°C, mutant enzyme H689Q or mutant H689A Phanerodontia chrysosporium
0.004
-
cellobiose pH 4.5, 23°C, mutant enzyme H689N or mutant H689Q Phanerodontia chrysosporium
0.004
-
cellobiose pH 4.5, 23°C, mutant enzyme N732D Phanerodontia chrysosporium
0.005
-
lactose pH 4.5, 23°C, mutant enzyme N732D Phanerodontia chrysosporium
0.009
-
cellobiose pH 4.5, 23°C, mutant enzyme H689E Phanerodontia chrysosporium
0.21
-
cellobiose pH 4.5, 23°C, mutant enzyme N732E Phanerodontia chrysosporium
0.3
-
lactose pH 4.5, 23°C, mutant enzyme N732E Phanerodontia chrysosporium
0.4
-
cellobiose pH 4.5, 23°C, mutant enzyme N732A Phanerodontia chrysosporium
0.7
-
lactose pH 4.5, 23°C, mutant enzyme N732A Phanerodontia chrysosporium
1
-
cellobiose pH 4.5, 23°C, mutant enzyme N732Q Phanerodontia chrysosporium
1.2
-
lactose pH 4.5, 23°C, mutant enzyme N732Q Phanerodontia chrysosporium
1.7
-
lactose pH 4.5, 23°C, mutant enzyme N732H Phanerodontia chrysosporium
2.7
-
cellobiose pH 4.5, 23°C, mutant enzyme N732H Phanerodontia chrysosporium
14.3
-
lactose pH 4.5, 23°C, recombinant wild-type enzyme Phanerodontia chrysosporium
15.5
-
cellobiose pH 4.5, 23°C, recombinant wild-type enzyme Phanerodontia chrysosporium

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
3
-
cellobiose + cytochrome c, highest activity at pH 3, decreasing as the pH increases, recombinant wild-type enzyme, mutant enzymes N732H, N732Q and N732A Phanerodontia chrysosporium
3 5 cellobiose + 2,6-dichlorophenol-indophenol, wild-type enzyme Phanerodontia chrysosporium
5
-
cellobiose + cytochrome c, mutant enzyme N732E Phanerodontia chrysosporium
6
-
cellobiose + cytochrome c, mutant enzyme N732D Phanerodontia chrysosporium
6.5 7 cellobiose + 2,6-dichlorophenol-indophenol, mutant enzyme N732E and N732D Phanerodontia chrysosporium

pH Range

pH Minimum pH Maximum Comment Organism
3 7 highest activity at pH 3, decreases when the pH increases to pH 7.0, cellobiose + 2,6-dichlorophenol-indophenol, to about 10% with recombinant wild-type enzyme, to about 25% with mutant enzymes N732Q, to about 40% with mutant enzyme N732A and to about 50% with mutant enzyme N732H Phanerodontia chrysosporium
3 5.5 highest activity at pH 3, decreases to 10-20% of the activity when the pH increases to pH 5.5, cellobiose + cytochrome c recombinant wild-type enzyme, mutant enzymes N732H, N732Q and N732A Phanerodontia chrysosporium
4 6 pH 5.0: about 35% of maximal activity, pH 7.0: about 20% of maximal activity, cellobiose + cytochrome c, mutant enzyme N732E Phanerodontia chrysosporium
5 7 pH 5.0: about 40% of maximal activity, pH 7.0: about 20% of maximal activity, cellobiose + cytochrome c, mutant enzyme N732D Phanerodontia chrysosporium
5.5 7.5 pH 5.0: about 45% of maximal activity, pH 7.0: about 65% of maximal activity, cellobiose + 2,6-dichlorophenol-indophenol, mutant enzyme N732E Phanerodontia chrysosporium
5.5 8 pH 5.5: about 50% of maximal activity, pH 8.0: about 60% of maximal activity, cellobiose + 2,6-dichlorophenol-indophenol, mutant enzyme N732D Phanerodontia chrysosporium

Cofactor

Cofactor Comment Organism Structure
cytochrome c the enzyme is an extracellular flavocytochrome Phanerodontia chrysosporium
flavin the enzyme is an extracellular flavocytochrome Phanerodontia chrysosporium