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Literature summary extracted from

  • Wohlschlager, L.; Csarman, F.; Chang, H.; Fitz, E.; Seiboth, B.; Ludwig, R.
    Heterologous expression of Phanerochaete chrysosporium cellobiose dehydrogenase in Trichoderma reesei (2021), Microb. Cell Fact., 20, 2 .
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
1.1.99.18 biofuel production key enzyme in biofuel cells Phanerodontia chrysosporium
1.1.99.18 diagnostics key enzyme in biosensors Phanerodontia chrysosporium

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.99.18 expression in Trichoderma reesei. Heterologous production of the enzyme (PcCDH) is faster and the yield higher than secretion by Phanerochaete chrysosporium. It does not need a cellulose-based medium that impedes efficient production and purification of the enzyme (PcCDH) by binding to the polysaccharide. The obtained high uniformity of Phanerochaete chrysosporium CDHTr glycoforms is very useful to investigate electron transfer characteristics in biosensors and biofuel cells, which are depending on the spatial restrictions inflicted by high-mannose N-glycan trees. The determined catalytic and electrochemical properties of PcCDHTr are very similar to those of PcCDH and the FAD cofactor occupancy is good, which advocates Trichoderma reesei as expression host for engineered PcCDH for biosensors and biofuel cells Phanerodontia chrysosporium

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.99.18 0.00036
-
 cytochrome c pH 4.0, 30°C Phanerodontia chrysosporium
1.1.99.18 0.00431
-
 2,6-dichlorophenolindophenol pH 4.5, 30°C Phanerodontia chrysosporium
1.1.99.18 0.08
-
 cellobiose pH 4.5, 30°C, acceptor: 2,6-dichlorophenolindophenol Phanerodontia chrysosporium
1.1.99.18 1.42
-
 lactose pH 4.5, 30°C, acceptor: 2,6-dichlorophenolindophenol Phanerodontia chrysosporium
1.1.99.18 2109
-
 D-glucose pH 4.5, 30°C, acceptor: 2,6-dichlorophenolindophenol Phanerodontia chrysosporium

Organism

EC Number Organism UniProt Comment Textmining
1.1.99.18 Phanerodontia chrysosporium Q01738
-
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
1.1.99.18 glycoprotein high mannose-type N-glycans in the enzyme from Phanerochaete chrysosporium. Only single N-acetyl-D-glucosamine additions at the six potential N-glycosylation sites in the enzyme recombinantly expressed in Trichoderma reesei Phanerodontia chrysosporium

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.99.18 enzyme from Phanerochaete chrysosporium and enzyme recombinantly expressed in Trichoderma reesei Phanerodontia chrysosporium

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.99.18 12.7
-
 30°C, pH 4.5, substrates: lactose + cytochrome c Phanerodontia chrysosporium

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.99.18 cellobiose + 1,4-benzoquinone
-
 Phanerodontia chrysosporium cellobiono-1,5-lactone + reduced 1,4-benzoquinone
-
 ?
1.1.99.18 cellobiose + 2,6-dichlorophenolindophenol
-
 Phanerodontia chrysosporium cellobiono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
-
 ?
1.1.99.18 cellobiose + cytochrome c
-
 Phanerodontia chrysosporium cellobiono-1,5-lactone + reduced cytochrome c
-
 ?
1.1.99.18 D-glucose + 2,6-dichloroindophenol
-
 Phanerodontia chrysosporium D-glucono-1,5-lactone + reduced 2,6-dichloroindophenol
-
 ?
1.1.99.18 lactose + 2,6-dichloroindophenol
-
 Phanerodontia chrysosporium 4-O-(beta-D-galactopyranosyl)-D-glucono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.1.99.18 Cdh
-
 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 7.8
-
 D-glucose pH 4.5, 30°C, acceptor: 2,6-dichlorophenolindophenol Phanerodontia chrysosporium
1.1.99.18 13
-
 cytochrome c pH 4.0, 30°C Phanerodontia chrysosporium
1.1.99.18 24.4
-
 2,6-dichloroindophenol pH 4.5, 30°C Phanerodontia chrysosporium
1.1.99.18 27.8
-
 cellobiose pH 4.5, 30°C, acceptor: 2,6-dichlorophenolindophenol Phanerodontia chrysosporium
1.1.99.18 32.3
-
 lactose pH 4.5, 30°C, acceptor: 2,6-dichlorophenolindophenol Phanerodontia chrysosporium

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.99.18 4
-
 activity with cytochrome c or 1,4-benzoquinone as acceptor Phanerodontia chrysosporium
1.1.99.18 4.5
-
 activity with 2,6-dichlorophenolindophenol as acceptor Phanerodontia chrysosporium

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.99.18 2.5 5 pH 2.5: about 15% of maximal activity, pH 5.0: about 40% of maximal activity, activity with cytochrome c as acceptor, Phanerochaete chrysosporium enzyme recombinantly expressed in Trichoderma reesei Phanerodontia chrysosporium
1.1.99.18 2.5 5 pH 2.5: about 45% of maximal activity, pH 5.0: about 55% of maximal activity, activity with cytochrome c as acceptor, enzyme from Phanerochaete chrysosporium Phanerodontia chrysosporium
1.1.99.18 3 6 more than 50% activity in the pH range between pH 3.0-6.0, acceptor: 1,4-benzoquinone Phanerodontia chrysosporium
1.1.99.18 3.2 6.2 pH 3.2: about 30% of maximal activity, pH 6.2: about 45% of maximal activity, activity with 2,6-dichlorophenolindophenol as acceptor, Phanerochaete chrysosporium enzyme recombinantly expressed in Trichoderma reesei Phanerodontia chrysosporium
1.1.99.18 3.5 6 pH 3.5: about 35% of maximal activity, pH 6.0: about 40% of maximal activity, activity with 2,6-dichlorophenolindophenol as acceptor, enzyme from Phanerochaete chrysosporium Phanerodontia chrysosporium

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.99.18 FAD FAD cofactor of the enzyme recombinantly expressed in Trichoderma reesei occupancy is 70% Phanerodontia chrysosporium

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.99.18 0.004
-
 D-glucose pH 4.5, 30°C, acceptor: 2,6-dichlorophenolindophenol Phanerodontia chrysosporium
1.1.99.18 22.7
-
 lactose pH 4.5, 30°C, acceptor: 2,6-dichlorophenolindophenol Phanerodontia chrysosporium
1.1.99.18 340
-
 cellobiose pH 4.5, 30°C, acceptor: 2,6-dichlorophenolindophenol Phanerodontia chrysosporium
1.1.99.18 5655
-
 2,6-dichlorophenolindophenol pH 4.5, 30°C Phanerodontia chrysosporium
1.1.99.18 36143
-
 cytochrome c pH 4.0, 30°C Phanerodontia chrysosporium