Literature summary for 1.1.99.18 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 .

Search for more literature for 1.1.99.18

Application

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

Cloned(Commentary)

Cloned (Comment)	Organism
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

Cloned (Comment)

Organism

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]

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

Organism

Organism	UniProt	Comment	Textmining
Phanerodontia chrysosporium	Q01738	-	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
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)

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

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

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

Synonyms

Synonyms	Comment	Organism
Cdh	-	Phanerodontia chrysosporium

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
7.8	-	D-glucose	pH 4.5, 30°C, acceptor: 2,6-dichlorophenolindophenol	Phanerodontia chrysosporium
13	-	cytochrome c	pH 4.0, 30°C	Phanerodontia chrysosporium
24.4	-	2,6-dichloroindophenol	pH 4.5, 30°C	Phanerodontia chrysosporium
27.8	-	cellobiose	pH 4.5, 30°C, acceptor: 2,6-dichlorophenolindophenol	Phanerodontia chrysosporium
32.3	-	lactose	pH 4.5, 30°C, acceptor: 2,6-dichlorophenolindophenol	Phanerodontia chrysosporium

pH Optimum

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

pH Range

pH Minimum	pH Maximum	Comment	Organism
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
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
3	6	more than 50% activity in the pH range between pH 3.0-6.0, acceptor: 1,4-benzoquinone	Phanerodontia chrysosporium
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
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

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

kcat/KM [mM/s]

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