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Literature summary extracted from

  • Blair, D.E.; Hekmat, O.; Schuettelkopf, A.W.; Shrestha, B.; Tokuyasu, K.; Withers, S.G.; van Aalten, D.M.
    Structure and mechanism of chitin deacetylase from the fungal pathogen Colletotrichum lindemuthianum (2006), Biochemistry, 45, 9416-9426.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.1.41 expression of the C-terminally His6-tagged enzyme in Pichia pastoris and in Escherichia coli, the His-tag causes intramolecular interactions with the active site Colletotrichum lindemuthianum

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.5.1.41 purified recombinant His6-tagged enzyme expressed in Pichia pastoris, sitting drop vapour diffusion method, 20 mg/ml protein in 30 mM Tris-HCl, pH 8.0, is mixed with an equal volume of reservoir solution containing 30% w/v PEG 4000, 0.2 M ammonium acetate, and 0.1 M sodium acetate, pH 4.6, 20°C, 10 days, plate-like crystals, soaking of crystals in mother liquor with 10 mM ZnCl2, for 3 h, cryoprotection by 15% glycerol, flash freezing, and X-ray diffraction structure determination and analysis at 1.8 A resolution Colletotrichum lindemuthianum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.1.41 0.102
-
 2-acetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-bromoacetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-acetamido-2-deoxy-D-glucose pH 8.5, 30°C, continuous spectrophotometric assay Colletotrichum lindemuthianum
3.5.1.41 0.125
-
 GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNAc pH 8.5, 30°C, continuous spectrophotometric assay Colletotrichum lindemuthianum
3.5.1.41 0.14
-
 2-acetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-fluoroacetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-acetamido-2-deoxy-D-glucose pH 8.5, 30°C, continuous spectrophotometric assay Colletotrichum lindemuthianum
3.5.1.41 0.18
-
 2-acetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-chloroacetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-acetamido-2-deoxy-D-glucose pH 8.5, 30°C, continuous spectrophotometric assay Colletotrichum lindemuthianum
3.5.1.41 0.66
-
 GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNAc pH 8.5, 30°C, continuous spectrophotometric assay Colletotrichum lindemuthianum
3.5.1.41 4.3
-
 GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNAc pH 7.0, 37°C, fluorogenic labeling method assay Colletotrichum lindemuthianum
3.5.1.41 18
-
 GlcNAcbeta(1-4)GlcNAc pH 8.5, 30°C, continuous spectrophotometric assay Colletotrichum lindemuthianum

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.5.1.41 extracellular the enzyme is secreted Colletotrichum lindemuthianum
-
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.5.1.41 Zn2+ tightly bound to the conserved metal-binding triad consisting of His104, His108, and Asp50, structure analysis Colletotrichum lindemuthianum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.5.1.41 chitin + H2O Colletotrichum lindemuthianum
-
 chitosan + acetate
-
 ?
3.5.1.41 chitin + H2O Colletotrichum lindemuthianum UPS9
-
 chitosan + acetate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.5.1.41 Colletotrichum lindemuthianum Q6DWK3
-
-
3.5.1.41 Colletotrichum lindemuthianum UPS9 Q6DWK3
-
-

Reaction

EC Number Reaction Comment Organism Reaction ID
3.5.1.41 chitin + H2O = chitosan + acetate ligand binding to the enzyme, computational docking analysis, catalytic mechanism via a tetrahedral oxyanion intermediate, and active site structure with a conserved metal-binding triad consisting of His104, His108, and Asp50 Colletotrichum lindemuthianum
a

Source Tissue

EC Number Source Tissue Comment Organism Textmining

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.1.41 2-acetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-bromoacetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-acetamido-2-deoxy-D-glucose + H2O
-
 Colletotrichum lindemuthianum GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNbeta(1-4)GlcNAc + bromoacetate
-
 ?
3.5.1.41 2-acetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-bromoacetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-acetamido-2-deoxy-D-glucose + H2O
-
 Colletotrichum lindemuthianum UPS9 GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNbeta(1-4)GlcNAc + bromoacetate
-
 ?
3.5.1.41 2-acetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-chloroacetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-acetamido-2-deoxy-D-glucose + H2O
-
 Colletotrichum lindemuthianum GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNbeta(1-4)GlcNAc + chloroacetate
-
 ?
3.5.1.41 2-acetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-chloroacetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-acetamido-2-deoxy-D-glucose + H2O
-
 Colletotrichum lindemuthianum UPS9 GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNbeta(1-4)GlcNAc + chloroacetate
-
 ?
3.5.1.41 2-acetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-fluoroacetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-acetamido-2-deoxy-D-glucose + H2O
-
 Colletotrichum lindemuthianum GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNbeta(1-4)GlcNAc + fluoroacetate
-
 ?
3.5.1.41 chitin + H2O
-
 Colletotrichum lindemuthianum chitosan + acetate
-
 ?
3.5.1.41 chitin + H2O the enzyme is an endo-chitin de-N-acetylase Colletotrichum lindemuthianum chitosan + acetate
-
 ?
3.5.1.41 chitin + H2O
-
 Colletotrichum lindemuthianum UPS9 chitosan + acetate
-
 ?
3.5.1.41 chitin + H2O the enzyme is an endo-chitin de-N-acetylase Colletotrichum lindemuthianum UPS9 chitosan + acetate
-
 ?
3.5.1.41 GlcNAcbeta(1-4)GlcNAc + H2O
-
 Colletotrichum lindemuthianum ?
-
 ?
3.5.1.41 GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNAc + H2O
-
 Colletotrichum lindemuthianum ?
-
 ?
3.5.1.41 GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNAc + H2O
-
 Colletotrichum lindemuthianum GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNbeta(1-4)GlcNAc + acetate
-
 ?
3.5.1.41 additional information binding study of an N-acetylglucosaminyl trimer to the enzyme, computational docking Colletotrichum lindemuthianum ?
-
 ?
3.5.1.41 additional information binding study of an N-acetylglucosaminyl trimer to the enzyme, computational docking Colletotrichum lindemuthianum UPS9 ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.5.1.41 CDA
-
 Colletotrichum lindemuthianum
3.5.1.41 More the enzyme belongs to the carbohydrate esterase family CE 4 Colletotrichum lindemuthianum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.1.41 30
-
 assay at, Escherichia coli expressed recombinant enzyme, continuous spectrophotometric assay Colletotrichum lindemuthianum
3.5.1.41 37
-
 assay at, Pichia pastoris expressed recombinant enzyme, fluorogenic labeling method assay Colletotrichum lindemuthianum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5.1.41 1.53
-
 GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNAc pH 7.0, 37°C, fluorogenic labeling method assay Colletotrichum lindemuthianum
3.5.1.41 6
-
 GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNAc pH 8.5, 30°C, continuous spectrophotometric assay Colletotrichum lindemuthianum
3.5.1.41 7
-
 GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNAc pH 8.5, 30°C, continuous spectrophotometric assay Colletotrichum lindemuthianum
3.5.1.41 11
-
 GlcNAcbeta(1-4)GlcNAc pH 8.5, 30°C, continuous spectrophotometric assay Colletotrichum lindemuthianum
3.5.1.41 31
-
 2-acetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-bromoacetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-acetamido-2-deoxy-D-glucose pH 8.5, 30°C, continuous spectrophotometric assay Colletotrichum lindemuthianum
3.5.1.41 47
-
 2-acetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-chloroacetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-acetamido-2-deoxy-D-glucose pH 8.5, 30°C, continuous spectrophotometric assay Colletotrichum lindemuthianum
3.5.1.41 68
-
 2-acetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-fluoroacetamido-2-deoxy-beta-D-glucopyranosyl-(1-4)-2-acetamido-2-deoxy-D-glucose pH 8.5, 30°C, continuous spectrophotometric assay Colletotrichum lindemuthianum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.1.41 7
-
 assay at, Pichia pastoris expressed recombinant enzyme, fluorogenic labeling method assay Colletotrichum lindemuthianum
3.5.1.41 8.5
-
 assay at, Escherichia coli expressed recombinant enzyme, continuous spectrophotometric assay Colletotrichum lindemuthianum