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Literature summary 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)

EC Number Cloned (Comment) Organism
1.1.99.18
-
 Phanerodontia chrysosporium

Protein Variants

EC Number Protein Variants Comment Organism
1.1.99.18 H689A more than 1000fold lower turnover value, Km-value for cellobiose and lactose is similar to that of the wild-type enzyme Phanerodontia chrysosporium
1.1.99.18 H689E more than 1000fold lower turnover value, Km-value for cellobiose and lactose is similar to that of the wild-type enzyme Phanerodontia chrysosporium
1.1.99.18 H689N more than 1000fold lower turnover value, Km-value for cellobiose and lactose is similar to that of the wild-type enzyme Phanerodontia chrysosporium
1.1.99.18 H689Q more than 1000fold lower turnover value, Km-value for cellobiose and lactose is similar to that of the wild-type enzyme Phanerodontia chrysosporium
1.1.99.18 H689V more than 1000fold lower turnover value, Km-value for cellobiose and lactose is similar to that of the wild-type enzyme Phanerodontia chrysosporium
1.1.99.18 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
1.1.99.18 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
1.1.99.18 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
1.1.99.18 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
1.1.99.18 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.99.18 0.016
-
 cellobiose pH 4.5, 23°C, recombinant wild-type enzyme Phanerodontia chrysosporium
1.1.99.18 0.017
-
 cellobiose pH 4.5, 23°C, mutant enzyme N732A Phanerodontia chrysosporium
1.1.99.18 0.019
-
 cellobiose pH 4.5, 23°C, mutant enzyme H689N Phanerodontia chrysosporium
1.1.99.18 0.021
-
 cellobiose pH 4.5, 23°C, mutant enzyme H689Q or mutant enzyme H689A Phanerodontia chrysosporium
1.1.99.18 0.022
-
 cellobiose pH 4.5, 23°C, mutant enzyme H689E Phanerodontia chrysosporium
1.1.99.18 0.031
-
 cellobiose pH 4.5, 23°C, mutant enzyme H689V Phanerodontia chrysosporium
1.1.99.18 0.04
-
 cellobiose pH 4.5, 23°C, mutant enzyme N732H Phanerodontia chrysosporium
1.1.99.18 0.041
-
 cellobiose pH 4.5, 23°C, mutant enzyme N732Q Phanerodontia chrysosporium
1.1.99.18 0.17
-
 cellobiose pH 4.5, 23°C, mutant enzyme N732D Phanerodontia chrysosporium
1.1.99.18 0.23
-
 cellobiose pH 4.5, 23°C, mutant enzyme N732E Phanerodontia chrysosporium
1.1.99.18 0.27
-
 lactose pH 4.5, 23°C, recombinant wild-type enzyme Phanerodontia chrysosporium
1.1.99.18 0.28
-
 lactose pH 4.5, 23°C, mutant enzyme H689A Phanerodontia chrysosporium
1.1.99.18 0.29
-
 lactose pH 4.5, 23°C, mutant enzyme H689Q Phanerodontia chrysosporium
1.1.99.18 1.2
-
 lactose pH 4.5, 23°C, mutant enzyme N732A Phanerodontia chrysosporium
1.1.99.18 2.3
-
 lactose pH 4.5, 23°C, mutant enzyme N732H Phanerodontia chrysosporium
1.1.99.18 4
-
 lactose pH 4.5, 23°C, mutant enzyme N732Q Phanerodontia chrysosporium
1.1.99.18 11.4
-
 lactose pH 4.5, 23°C, mutant enzyme N732D Phanerodontia chrysosporium
1.1.99.18 16.6
-
 lactose pH 4.5, 23°C, mutant enzyme N732E Phanerodontia chrysosporium

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.1.99.18 extracellular
-
 Phanerodontia chrysosporium
-
-

Organism

EC Number Organism UniProt Comment Textmining
1.1.99.18 Phanerodontia chrysosporium
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.99.18 cellobiose + 2,6-dichlorophenol-indophenol
-
 Phanerodontia chrysosporium cellobiono-1,5-lactone + reduced 2,6-dichlorophenol-indophenol
-
 ?
1.1.99.18 cellobiose + cytochrome c
-
 Phanerodontia chrysosporium cellobiono-1,5-lactone + reduced cytochrome c
-
 ?
1.1.99.18 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

EC Number Synonyms Comment Organism
1.1.99.18 Cdh
-
 Phanerodontia chrysosporium
1.1.99.18 cellobiose dehydrogenase
-
 Phanerodontia chrysosporium

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.99.18 0.002
-
 cellobiose pH 4.5, 23°C, mutant enzyme H689V Phanerodontia chrysosporium
1.1.99.18 0.003
-
 cellobiose pH 4.5, 23°C, mutant enzyme H689A Phanerodontia chrysosporium
1.1.99.18 0.003
-
 lactose pH 4.5, 23°C, mutant enzyme H689Q or mutant H689A Phanerodontia chrysosporium
1.1.99.18 0.004
-
 cellobiose pH 4.5, 23°C, mutant enzyme H689N or mutant H689Q Phanerodontia chrysosporium
1.1.99.18 0.004
-
 cellobiose pH 4.5, 23°C, mutant enzyme N732D Phanerodontia chrysosporium
1.1.99.18 0.005
-
 lactose pH 4.5, 23°C, mutant enzyme N732D Phanerodontia chrysosporium
1.1.99.18 0.009
-
 cellobiose pH 4.5, 23°C, mutant enzyme H689E Phanerodontia chrysosporium
1.1.99.18 0.21
-
 cellobiose pH 4.5, 23°C, mutant enzyme N732E Phanerodontia chrysosporium
1.1.99.18 0.3
-
 lactose pH 4.5, 23°C, mutant enzyme N732E Phanerodontia chrysosporium
1.1.99.18 0.4
-
 cellobiose pH 4.5, 23°C, mutant enzyme N732A Phanerodontia chrysosporium
1.1.99.18 0.7
-
 lactose pH 4.5, 23°C, mutant enzyme N732A Phanerodontia chrysosporium
1.1.99.18 1
-
 cellobiose pH 4.5, 23°C, mutant enzyme N732Q Phanerodontia chrysosporium
1.1.99.18 1.2
-
 lactose pH 4.5, 23°C, mutant enzyme N732Q Phanerodontia chrysosporium
1.1.99.18 1.7
-
 lactose pH 4.5, 23°C, mutant enzyme N732H Phanerodontia chrysosporium
1.1.99.18 2.7
-
 cellobiose pH 4.5, 23°C, mutant enzyme N732H Phanerodontia chrysosporium
1.1.99.18 14.3
-
 lactose pH 4.5, 23°C, recombinant wild-type enzyme Phanerodontia chrysosporium
1.1.99.18 15.5
-
 cellobiose pH 4.5, 23°C, recombinant wild-type enzyme Phanerodontia chrysosporium

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.99.18 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
1.1.99.18 3 5 cellobiose + 2,6-dichlorophenol-indophenol, wild-type enzyme Phanerodontia chrysosporium
1.1.99.18 5
-
 cellobiose + cytochrome c, mutant enzyme N732E Phanerodontia chrysosporium
1.1.99.18 6
-
 cellobiose + cytochrome c, mutant enzyme N732D Phanerodontia chrysosporium
1.1.99.18 6.5 7 cellobiose + 2,6-dichlorophenol-indophenol, mutant enzyme N732E and N732D Phanerodontia chrysosporium

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.99.18 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
1.1.99.18 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
1.1.99.18 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
1.1.99.18 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
1.1.99.18 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
1.1.99.18 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

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