Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli strain DH5alpha | Trichoderma reesei |
gene cel7A, recombinant expression of wild-type and mutant W40A enzymes under the control of its own promoter in Trichoderma reesei strain ALKO 3413 lacking the genes encoding for endogenous Cel7A and Cel7B | Trichoderma reesei |
Protein Variants | Comment | Organism |
---|---|---|
W40A | mutation of Trp40 at the entrance of the catalytic tunnel drastically decreases the ability to degrade crystalline cellulose. Comparison of activities of the wild-type and mutant W40A enzymes (with and without the cellulose-binding domain) for various substrates, overview | Trichoderma reesei |
W40A | the mutation causes a loss of crystalline cellulose-degrading ability. The mutant shows reduced specific activity for crystalline cellulose and diffused the cellulose chain from the entrance of the active site tunnel | Trichoderma reesei |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | 4-methylumbelliferyl beta-D-lactoside substrate inhibition | Trichoderma reesei |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetic | Trichoderma reesei | |
0.293 | - |
4-methylumbelliferyl beta-D-lactoside | pH 5.0, 27°C, recombinant mutant W40A enzyme catalytic domain | Trichoderma reesei | |
0.318 | - |
4-methylumbelliferyl beta-D-lactopyranoside | mutant enzyme W40A, at pH 5.0 and 27°C | Trichoderma reesei | |
0.318 | - |
4-methylumbelliferyl beta-D-lactoside | pH 5.0, 27°C, recombinant mutant W40A enzyme | Trichoderma reesei | |
0.335 | - |
4-methylumbelliferyl beta-D-lactoside | pH 5.0, 27°C, recombinant wild-type enzyme catalytic domain | Trichoderma reesei | |
0.358 | - |
4-methylumbelliferyl beta-D-lactoside | pH 5.0, 27°C, recombinant wild-type enzyme | Trichoderma reesei | |
0.358 | - |
4-methylumbelliferyl beta-D-lactopyranoside | wild type enzyme, at pH 5.0 and 27°C | Trichoderma reesei |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Trichoderma reesei | the enzyme has the ability to degrade highly crystalline cellulose. The catalytic domain and the cellulose-binding domain are both necessary for full activity on crystalline substrates | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Trichoderma reesei | - |
- |
- |
Trichoderma reesei | G0RVK1 | gene cel7A | - |
Trichoderma reesei ALKO 3413 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Bio-Gel P-6 gel filtration, DEAE-Sepharose column chromatography and phenyl Sepharose column chromatography | Trichoderma reesei |
recombinant -type and mutant W40A enzymes from Trichoderma reesei strain ALKO 3413 culture filtrate by gel filtration, anion exchange, hydrophobic interaction, and affinity chromatography | Trichoderma reesei |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-methylumbelliferyl beta-D-cellobioside + H2O | - |
Trichoderma reesei | 4-methylumbelliferone + cellobiose | - |
? | |
4-methylumbelliferyl beta-D-cellobioside + H2O | - |
Trichoderma reesei ALKO 3413 | 4-methylumbelliferone + cellobiose | - |
? | |
4-methylumbelliferyl beta-D-lactopyranoside + H2O | - |
Trichoderma reesei | 4-methylumbelliferone + lactose | - |
? | |
4-methylumbelliferyl beta-D-lactopyranoside + H2O | - |
Trichoderma reesei ALKO 3413 | 4-methylumbelliferone + lactose | - |
? | |
4-methylumbelliferyl beta-D-lactoside + H2O | - |
Trichoderma reesei | 4-methylumbelliferol + D-lactose | - |
? | |
crystalline cellulose + H2O | the enzyme shows processive hydrolysis by the catalytic domain and a sliding movement of single enzyme molecules on a highly crystalline cellulose surface, the cellulose-binding domain is unnecessary for the sliding movement, real-time monitoring by high-speed atomic force microscope | Trichoderma reesei | ? | - |
? | |
highly crystalline cellulose + H2O | - |
Trichoderma reesei | cellobiose | - |
? | |
highly crystalline cellulose + H2O | - |
Trichoderma reesei ALKO 3413 | cellobiose | - |
? | |
additional information | the enzyme has the ability to degrade highly crystalline cellulose. The catalytic domain and the cellulose-binding domain are both necessary for full activity on crystalline substrates | Trichoderma reesei | ? | - |
? | |
additional information | comparison of activities of the wild-type and mutangt W40A enzymes (with and without the cellulose-binding domain) for various substrates, overview | Trichoderma reesei | ? | - |
? | |
phosphoric acid-swollen cellulose + H2O | - |
Trichoderma reesei | cellobiose | - |
? | |
phosphoric acid-swollen cellulose + H2O | - |
Trichoderma reesei ALKO 3413 | cellobiose | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | three-dimensional structure of the catalytic domain of TrCel7A cellobiohydrolase, overview | Trichoderma reesei |
Synonyms | Comment | Organism |
---|---|---|
Cel7A | - |
Trichoderma reesei |
cellobiohydrolase | - |
Trichoderma reesei |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.33 | - |
4-methylumbelliferyl beta-D-lactopyranoside | mutant enzyme W40A, at pH 5.0 and 27°C | Trichoderma reesei | |
0.33 | - |
4-methylumbelliferyl beta-D-lactoside | pH 5.0, 27°C, recombinant mutant W40A enzyme | Trichoderma reesei | |
0.361 | - |
4-methylumbelliferyl beta-D-lactoside | pH 5.0, 27°C, recombinant mutant W40A enzyme catalytic domain | Trichoderma reesei | |
0.415 | - |
4-methylumbelliferyl beta-D-lactoside | pH 5.0, 27°C, recombinant wild-type enzyme catalytic domain | Trichoderma reesei | |
0.49 | - |
4-methylumbelliferyl beta-D-lactopyranoside | wild type enzyme, at pH 5.0 and 27°C | Trichoderma reesei | |
0.492 | - |
4-methylumbelliferyl beta-D-lactoside | pH 5.0, 27°C, recombinant wild-type enzyme | Trichoderma reesei |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5 | 6 | assay at | Trichoderma reesei |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the glycoside hydrolase family 7, GH7 | Trichoderma reesei |
malfunction | mutation of Trp40 at the entrance of the catalytic tunnel drastically decreases the ability to degrade crystalline cellulose. Inactive mutant enzyme does not slide on a cellulose surface | Trichoderma reesei |
additional information | three-dimensional structure of the catalytic domain of TrCel7A cellobiohydrolase, Trp40 is involved in recruiting individual substrate chains into the active site tunnel to initiate processive hydrolysis, molecular dynamics simulation study, overview. The reducing end glucose unit is effectively loaded into the active site ofWTcat, but not into that of W40Acat, when the simulation is started from subsite-7 | Trichoderma reesei |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.03 | - |
4-methylumbelliferyl beta-D-lactopyranoside | mutant enzyme W40A, at pH 5.0 and 27°C | Trichoderma reesei | |
1.03 | - |
4-methylumbelliferyl beta-D-lactoside | pH 5.0, 27°C, recombinant mutant W40A enzyme | Trichoderma reesei | |
1.23 | - |
4-methylumbelliferyl beta-D-lactoside | pH 5.0, 27°C, recombinant mutant W40A enzyme catalytic domain | Trichoderma reesei | |
1.23 | - |
4-methylumbelliferyl beta-D-lactoside | pH 5.0, 27°C, recombinant wild-type enzyme catalytic domain | Trichoderma reesei | |
1.37 | - |
4-methylumbelliferyl beta-D-lactoside | pH 5.0, 27°C, recombinant wild-type enzyme | Trichoderma reesei | |
1.37 | - |
4-methylumbelliferyl beta-D-lactopyranoside | wild type enzyme, at pH 5.0 and 27°C | Trichoderma reesei |