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Literature summary extracted from

  • Nakamura, A.; Tsukada, T.; Auer, S.; Furuta, T.; Wada, M.; Koivula, A.; Igarashi, K.; Samejima, M.
    The tryptophan residue at the active site tunnel entrance of Trichoderma reesei cellobiohydrolase Cel7A is important for initiation of degradation of crystalline cellulose (2013), J. Biol. Chem., 288, 13503-13510.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.176 expressed in Escherichia coli strain DH5alpha Trichoderma reesei
3.2.1.176 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

EC Number Protein Variants Comment Organism
3.2.1.176 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
3.2.1.176 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

EC Number Inhibitors Comment Organism Structure
3.2.1.176 additional information 4-methylumbelliferyl beta-D-lactoside substrate inhibition Trichoderma reesei

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.176 additional information
-
 additional information Michaelis-Menten kinetic Trichoderma reesei
3.2.1.176 0.293
-
 4-methylumbelliferyl beta-D-lactoside pH 5.0, 27°C, recombinant mutant W40A enzyme catalytic domain Trichoderma reesei
3.2.1.176 0.318
-
 4-methylumbelliferyl beta-D-lactopyranoside mutant enzyme W40A, at pH 5.0 and 27°C Trichoderma reesei
3.2.1.176 0.318
-
 4-methylumbelliferyl beta-D-lactoside pH 5.0, 27°C, recombinant mutant W40A enzyme Trichoderma reesei
3.2.1.176 0.335
-
 4-methylumbelliferyl beta-D-lactoside pH 5.0, 27°C, recombinant wild-type enzyme catalytic domain Trichoderma reesei
3.2.1.176 0.358
-
 4-methylumbelliferyl beta-D-lactoside pH 5.0, 27°C, recombinant wild-type enzyme Trichoderma reesei
3.2.1.176 0.358
-
 4-methylumbelliferyl beta-D-lactopyranoside wild type enzyme, at pH 5.0 and 27°C Trichoderma reesei

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.1.176 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

EC Number Organism UniProt Comment Textmining
3.2.1.176 Trichoderma reesei
-
-
-
3.2.1.176 Trichoderma reesei G0RVK1 gene cel7A
-
3.2.1.176 Trichoderma reesei ALKO 3413
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.176 Bio-Gel P-6 gel filtration, DEAE-Sepharose column chromatography and phenyl Sepharose column chromatography Trichoderma reesei
3.2.1.176 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 and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.176 4-methylumbelliferyl beta-D-cellobioside + H2O
-
 Trichoderma reesei 4-methylumbelliferone + cellobiose
-
 ?
3.2.1.176 4-methylumbelliferyl beta-D-cellobioside + H2O
-
 Trichoderma reesei ALKO 3413 4-methylumbelliferone + cellobiose
-
 ?
3.2.1.176 4-methylumbelliferyl beta-D-lactopyranoside + H2O
-
 Trichoderma reesei 4-methylumbelliferone + lactose
-
 ?
3.2.1.176 4-methylumbelliferyl beta-D-lactopyranoside + H2O
-
 Trichoderma reesei ALKO 3413 4-methylumbelliferone + lactose
-
 ?
3.2.1.176 4-methylumbelliferyl beta-D-lactoside + H2O
-
 Trichoderma reesei 4-methylumbelliferol + D-lactose
-
 ?
3.2.1.176 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 ?
-
 ?
3.2.1.176 highly crystalline cellulose + H2O
-
 Trichoderma reesei cellobiose
-
 ?
3.2.1.176 highly crystalline cellulose + H2O
-
 Trichoderma reesei ALKO 3413 cellobiose
-
 ?
3.2.1.176 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 ?
-
 ?
3.2.1.176 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 ?
-
 ?
3.2.1.176 phosphoric acid-swollen cellulose + H2O
-
 Trichoderma reesei cellobiose
-
 ?
3.2.1.176 phosphoric acid-swollen cellulose + H2O
-
 Trichoderma reesei ALKO 3413 cellobiose
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.176 More three-dimensional structure of the catalytic domain of TrCel7A cellobiohydrolase, overview Trichoderma reesei

Synonyms

EC Number Synonyms Comment Organism
3.2.1.176 Cel7A
-
 Trichoderma reesei
3.2.1.176 cellobiohydrolase
-
 Trichoderma reesei

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.176 0.33
-
 4-methylumbelliferyl beta-D-lactopyranoside mutant enzyme W40A, at pH 5.0 and 27°C Trichoderma reesei
3.2.1.176 0.33
-
 4-methylumbelliferyl beta-D-lactoside pH 5.0, 27°C, recombinant mutant W40A enzyme Trichoderma reesei
3.2.1.176 0.361
-
 4-methylumbelliferyl beta-D-lactoside pH 5.0, 27°C, recombinant mutant W40A enzyme catalytic domain Trichoderma reesei
3.2.1.176 0.415
-
 4-methylumbelliferyl beta-D-lactoside pH 5.0, 27°C, recombinant wild-type enzyme catalytic domain Trichoderma reesei
3.2.1.176 0.49
-
 4-methylumbelliferyl beta-D-lactopyranoside wild type enzyme, at pH 5.0 and 27°C Trichoderma reesei
3.2.1.176 0.492
-
 4-methylumbelliferyl beta-D-lactoside pH 5.0, 27°C, recombinant wild-type enzyme Trichoderma reesei

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.176 5 6 assay at Trichoderma reesei

General Information

EC Number General Information Comment Organism
3.2.1.176 evolution the enzyme belongs to the glycoside hydrolase family 7, GH7 Trichoderma reesei
3.2.1.176 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
3.2.1.176 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 [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.1.176 1.03
-
 4-methylumbelliferyl beta-D-lactopyranoside mutant enzyme W40A, at pH 5.0 and 27°C Trichoderma reesei
3.2.1.176 1.03
-
 4-methylumbelliferyl beta-D-lactoside pH 5.0, 27°C, recombinant mutant W40A enzyme Trichoderma reesei
3.2.1.176 1.23
-
 4-methylumbelliferyl beta-D-lactoside pH 5.0, 27°C, recombinant mutant W40A enzyme catalytic domain Trichoderma reesei
3.2.1.176 1.23
-
 4-methylumbelliferyl beta-D-lactoside pH 5.0, 27°C, recombinant wild-type enzyme catalytic domain Trichoderma reesei
3.2.1.176 1.37
-
 4-methylumbelliferyl beta-D-lactoside pH 5.0, 27°C, recombinant wild-type enzyme Trichoderma reesei
3.2.1.176 1.37
-
 4-methylumbelliferyl beta-D-lactopyranoside wild type enzyme, at pH 5.0 and 27°C Trichoderma reesei