EC Number | Cloned (Comment) | Organism |
---|---|---|
3.2.1.2 | expression of wild-type and mutants in Escherichia coli strains JM109 and JM105 | Glycine max |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
3.2.1.2 | purified recombinant mutant enzymes, mutant enzyme E186Q in complex with substrate maltopentaose, and mutant enzyme E380Q in complex with product maltose, hanging drop vapour diffusion method, 20 mg/ml protein in 45-50% w/v ammonium sulfate, 0.1 M sodium acetate, pH 5.4, 1 mM EDTA, and 18 mM 2-methyl-2,4-pentanediol, equilibration against 1 ml mother liquor, 4°C, soaking of crystals in 30 mM ligand solution, cryoprotection of crystals with 30% v/v glycerol in crystallization solution, X-ray diffraction structure determination and analysis at 1.6 and 1.9 A resolution, respectively, active site structure modelling | Glycine max |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
3.2.1.2 | E186Q | site-directed mutagenesis, mutation of catalytic residue, the mutant shows 16000fold decreased activity compared to the wild-type enzyme | Glycine max |
3.2.1.2 | E380Q | site-directed mutagenesis, mutation of catalytic residue, the mutant shows 37000fold decreased activity compared to the wild-type enzyme | Glycine max |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.2.1.2 | 1.94 | - |
maltopentaose | pH 5.4, 37°C, wild-type enzyme | Glycine max | |
3.2.1.2 | 2.02 | - |
maltopentaose | pH 5.4, 37°C, mutant E380Q | Glycine max | |
3.2.1.2 | 2.15 | - |
maltopentaose | pH 5.4, 37°C, mutant E186Q | Glycine max |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.2.1.2 | amylopectin + H2O | Glycine max | - |
maltose + ? | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
3.2.1.2 | Glycine max | P10538 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
3.2.1.2 | recombinant wild-type and mutants from Escherichia coli | Glycine max |
EC Number | Reaction | Comment | Organism | Reaction ID |
---|---|---|---|---|
3.2.1.2 | (alpha-D-glucopyranosyl-(1-4))n-alpha-D-glucopyranose + H2O = (alpha-D-glucopyranosyl-(1-4))n-2-alpha-D-glucopyranose + alpha-D-glucopyranosyl-(1-4)-beta-D-glucopyranose | roles of Glu186 and Glu380 as general acid and general base catalyst in the catalytic reaction, substrate binding and reaction mechanism | Glycine max |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
3.2.1.2 | additional information | - |
- |
Glycine max |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.2.1.2 | amylopectin + H2O | - |
Glycine max | maltose + ? | - |
? | |
3.2.1.2 | amylopectin + H2O | from potato | Glycine max | maltose + ? | - |
? | |
3.2.1.2 | maltopentaose + H2O | substrate/product binding structure, sugar subsite conformations, overview | Glycine max | 2 maltose + D-glucose | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
3.2.1.2 | More | structure analysis, intramolecular hydrogen bond interactions | Glycine max |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
3.2.1.2 | SBA | - |
Glycine max |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
3.2.1.2 | 37 | - |
assay at | Glycine max |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.2.1.2 | 0.034 | - |
maltopentaose | pH 5.4, 37°C, mutant E380Q | Glycine max | |
3.2.1.2 | 0.08 | - |
maltopentaose | pH 5.4, 37°C, mutant E186Q | Glycine max | |
3.2.1.2 | 1280 | - |
maltopentaose | pH 5.4, 37°C, wild-type enzyme | Glycine max |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
3.2.1.2 | 5.4 | - |
assay at | Glycine max |