Cloned (Comment) | Organism |
---|---|
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 (Comment) | Organism |
---|---|
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] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
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 | |
0.125 | - |
GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNAc | pH 8.5, 30°C, continuous spectrophotometric assay | Colletotrichum lindemuthianum | |
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 | |
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 | |
0.66 | - |
GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNAc | pH 8.5, 30°C, continuous spectrophotometric assay | Colletotrichum lindemuthianum | |
4.3 | - |
GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNAc | pH 7.0, 37°C, fluorogenic labeling method assay | Colletotrichum lindemuthianum | |
18 | - |
GlcNAcbeta(1-4)GlcNAc | pH 8.5, 30°C, continuous spectrophotometric assay | Colletotrichum lindemuthianum |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
extracellular | the enzyme is secreted | Colletotrichum lindemuthianum | - |
- |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | tightly bound to the conserved metal-binding triad consisting of His104, His108, and Asp50, structure analysis | Colletotrichum lindemuthianum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
chitin + H2O | Colletotrichum lindemuthianum | - |
chitosan + acetate | - |
? | |
chitin + H2O | Colletotrichum lindemuthianum UPS9 | - |
chitosan + acetate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Colletotrichum lindemuthianum | Q6DWK3 | - |
- |
Colletotrichum lindemuthianum UPS9 | Q6DWK3 | - |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
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 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
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 | - |
? | |
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 | - |
? | |
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 | - |
? | |
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 | - |
? | |
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 | - |
? | |
chitin + H2O | - |
Colletotrichum lindemuthianum | chitosan + acetate | - |
? | |
chitin + H2O | the enzyme is an endo-chitin de-N-acetylase | Colletotrichum lindemuthianum | chitosan + acetate | - |
? | |
chitin + H2O | - |
Colletotrichum lindemuthianum UPS9 | chitosan + acetate | - |
? | |
chitin + H2O | the enzyme is an endo-chitin de-N-acetylase | Colletotrichum lindemuthianum UPS9 | chitosan + acetate | - |
? | |
GlcNAcbeta(1-4)GlcNAc + H2O | - |
Colletotrichum lindemuthianum | ? | - |
? | |
GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNAc + H2O | - |
Colletotrichum lindemuthianum | ? | - |
? | |
GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNAc + H2O | - |
Colletotrichum lindemuthianum | GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNbeta(1-4)GlcNAc + acetate | - |
? | |
additional information | binding study of an N-acetylglucosaminyl trimer to the enzyme, computational docking | Colletotrichum lindemuthianum | ? | - |
? | |
additional information | binding study of an N-acetylglucosaminyl trimer to the enzyme, computational docking | Colletotrichum lindemuthianum UPS9 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
CDA | - |
Colletotrichum lindemuthianum |
More | the enzyme belongs to the carbohydrate esterase family CE 4 | Colletotrichum lindemuthianum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at, Escherichia coli expressed recombinant enzyme, continuous spectrophotometric assay | Colletotrichum lindemuthianum |
37 | - |
assay at, Pichia pastoris expressed recombinant enzyme, fluorogenic labeling method assay | Colletotrichum lindemuthianum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.53 | - |
GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNAc | pH 7.0, 37°C, fluorogenic labeling method assay | Colletotrichum lindemuthianum | |
6 | - |
GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNAc | pH 8.5, 30°C, continuous spectrophotometric assay | Colletotrichum lindemuthianum | |
7 | - |
GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNAcbeta(1-4)GlcNAc | pH 8.5, 30°C, continuous spectrophotometric assay | Colletotrichum lindemuthianum | |
11 | - |
GlcNAcbeta(1-4)GlcNAc | pH 8.5, 30°C, continuous spectrophotometric assay | Colletotrichum lindemuthianum | |
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 | |
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 | |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at, Pichia pastoris expressed recombinant enzyme, fluorogenic labeling method assay | Colletotrichum lindemuthianum |
8.5 | - |
assay at, Escherichia coli expressed recombinant enzyme, continuous spectrophotometric assay | Colletotrichum lindemuthianum |