Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli strain JM105 | Glycine max |
Crystallization (Comment) | Organism |
---|---|
purified recombinant T342 mutant enzymes, hanging drop vapour diffusion method, 0.005 ml of 10 mg/ml protein solution is mixed with 0.005 ml of reservoir solution containing 40-50% w/v ammonium sulfate, 1 mM EDTA, 18 mM 2-methyl-2,4-pentanediol, and 0.1 M sodium acetate, pH 5.4, equilibration against 1 ml of reservoir solution, 4°C, gradual soaking of crystals in 0.1 M sodium acetate, pH 6.1, 50% w/v ammonium sulfate, 1 mM EDTA, 20 mM DTT, 0.3 M maltose, and 30% v/v glycerol, X-ray diffraction structure determination and analysis at 1.12-1.6 A resolution, active site structure modelling | Glycine max |
Protein Variants | Comment | Organism |
---|---|---|
T342A | site-directed mutagenesis, structural analysis of mutant active site conformation | Glycine max |
T342S | site-directed mutagenesis, structural analysis of mutant active site conformation | Glycine max |
T342V | site-directed mutagenesis, structural analysis of mutant active site conformation | Glycine max |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | - |
Glycine max | |
0.26 | - |
amylopectin | pH 5.4, 37°C, recombinant mutant T342A | Glycine max | |
0.39 | - |
amylopectin | pH 5.4, 37°C, recombinant mutant T342S | Glycine max | |
1.84 | - |
amylopectin | pH 5.4, 37°C, recombinant mutant T342V | Glycine max | |
1.94 | - |
amylopectin | pH 5.4, 37°C, recombinant wild-type enzyme | Glycine max |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Glycine max | P10538 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain JM105 | Glycine max |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
(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 | active site structure, carbohydrate binding subsites, role of a conformational change of the inner loop in the catalytic mechanism, T342 is involved, overview | Glycine max |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
amylopectin + H2O | from potato | Glycine max | maltose + ? | - |
? |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Glycine max |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.24 | - |
amylopectin | pH 5.4, 37°C, recombinant mutant T342V | Glycine max | |
0.75 | - |
amylopectin | pH 5.4, 37°C, recombinant mutant T342A | Glycine max | |
3.5 | - |
amylopectin | pH 5.4, 37°C, recombinant mutant T342S | Glycine max | |
98.4 | - |
amylopectin | pH 5.4, 37°C, recombinant mutant T342V | Glycine max | |
1280 | - |
amylopectin | pH 5.4, 37°C, recombinant wild-type enzyme | Glycine max |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5.4 | - |
- |
Glycine max |