Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli | Corynebacterium callunae |
Protein Variants | Comment | Organism |
---|---|---|
H334A | site-directed mutagenesis, catalytic efficiency in the phosphorolysis reaction is similar to the wild-type enzyme, 50fold reduced binding of alpha-D-glucose 1-phosphate compared to the wild-type enzyme | Corynebacterium callunae |
H334N | site-directed mutagenesis, 50fold reduced catalytic efficiency in the phosphorolysis reaction and highly reduced selectivity for alpha-D-glucose 1-phosphate, versus alpha-D-xylose 1-phosphate, in the synthesis reaction with 10fold reduced binding of alpha-D-glucose 1-phosphate compared to the wild-type enzyme | Corynebacterium callunae |
H334Q | site-directed mutagenesis, 150fold reduced catalytic efficiency in the phosphorolysis reaction and highly reduced selectivity for alpha-D-glucose 1-phosphate, versus alpha-D-xylose 1-phosphate, in the synthesis reaction with 10fold reduced binding of alpha-D-glucose 1-phosphate compared to the wild-type enzyme | Corynebacterium callunae |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
D-gluconic acid 1,5-lactone | - |
Corynebacterium callunae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | stopped-flow and steady-state kinetics, wild-type and mutant enzymes | Corynebacterium callunae | |
0.23 | - |
starch | pH 7.0, 30°C, recombinant wild-type enzyme, phosphorolysis | Corynebacterium callunae | |
0.25 | - |
starch | pH 7.0, 30°C, recombinant mutant H334A, phosphorolysis | Corynebacterium callunae | |
0.32 | - |
starch | pH 7.0, 30°C, recombinant mutant H334Q, phosphorolysis | Corynebacterium callunae | |
0.85 | - |
alpha-D-glucose 1-phosphate | pH 7.0, 30°C, recombinant wild-type enzyme, synthesis | Corynebacterium callunae | |
1 | - |
starch | pH 7.0, 30°C, recombinant mutant H334N, phosphorolysis | Corynebacterium callunae | |
1.1 | - |
alpha-D-glucose 1-phosphate | pH 7.0, 30°C, recombinant mutant H334Q, synthesis | Corynebacterium callunae | |
1.3 | - |
alpha-D-glucose 1-phosphate | pH 7.0, 30°C, recombinant mutant H334N, synthesis | Corynebacterium callunae | |
1.6 | - |
glycogen | pH 7.0, 30°C, recombinant wild-type enzyme and mutant H334A, phosphorolysis | Corynebacterium callunae | |
1.7 | - |
glycogen | pH 7.0, 30°C, recombinant mutant H334Q, phosphorolysis | Corynebacterium callunae | |
1.8 | - |
alpha-D-glucose 1-phosphate | pH 7.0, 30°C, recombinant mutant H334A, synthesis | Corynebacterium callunae | |
3.5 | - |
maltopentaose | pH 7.0, 30°C, recombinant mutant H334A, phosphorolysis | Corynebacterium callunae | |
3.8 | - |
maltopentaose | pH 7.0, 30°C, recombinant wild-type enzyme, phosphorolysis | Corynebacterium callunae | |
4.1 | - |
maltopentaose | pH 7.0, 30°C, recombinant mutant H334Q, phosphorolysis | Corynebacterium callunae | |
6.8 | - |
phosphate | pH 7.0, 30°C, recombinant wild-type enzyme, phosphorolysis | Corynebacterium callunae | |
12 | - |
phosphate | pH 7.0, 30°C, recombinant mutants H334A and H334Q, phosphorolysis | Corynebacterium callunae | |
13 | - |
phosphate | pH 7.0, 30°C, recombinant mutant H334N, phosphorolysis | Corynebacterium callunae | |
16 | - |
glycogen | pH 7.0, 30°C, recombinant mutant H334N, phosphorolysis | Corynebacterium callunae | |
103 | - |
maltopentaose | pH 7.0, 30°C, recombinant mutant H334N, phosphorolysis | Corynebacterium callunae |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Corynebacterium callunae | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli | Corynebacterium callunae |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
[(1->4)-alpha-D-glucosyl]n + phosphate = [(1->4)-alpha-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate | catalytic mechanism of alpha-retaining glucosyl transfer, His334 is involved in selectively stabilizing the oxycarbenium ion-like transition state of the reaction, ligand binding | Corynebacterium callunae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(1,4-alpha-D-glucosyl)n + phosphate | polysaccharide substrates are starch and glycogen, phosphorolysis is the preferred reaction, maltotetraose is the smallest substrate accepted | Corynebacterium callunae | (1,4-alpha-D-glucosyl)n-1 + alpha-D-glucose 1-phosphate | - |
r | |
(1,4-alpha-D-glucosyl)n-1 + alpha-D-xylose 1-phosphate | polysaccharide substrate is starch, very low activity | Corynebacterium callunae | alpha-D-xylose-(1,4-alpha-D-glucosyl)n + phosphate | - |
r | |
glycogen + glucose 1-phosphate | - |
Corynebacterium callunae | glycogen + phosphate | - |
? | |
maltopentaose + phosphate | - |
Corynebacterium callunae | maltotetraose + alpha-D-glucose 1-phosphate | - |
r | |
starch + glucose 1-phosphate | - |
Corynebacterium callunae | starch + phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
starch phosphorylase | - |
Corynebacterium callunae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Corynebacterium callunae |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00048 | - |
alpha-D-xylose 1-phosphate | pH 7.0, 30°C, recombinant mutant H334N, synthesis | Corynebacterium callunae | |
0.0026 | - |
alpha-D-xylose 1-phosphate | pH 7.0, 30°C, recombinant wild-type enzyme, synthesis | Corynebacterium callunae | |
0.0028 | - |
alpha-D-xylose 1-phosphate | pH 7.0, 30°C, recombinant mutant H334A, synthesis | Corynebacterium callunae | |
0.0066 | - |
alpha-D-xylose 1-phosphate | pH 7.0, 30°C, recombinant mutant H334Q, synthesis | Corynebacterium callunae | |
0.52 | - |
starch | pH 7.0, 30°C, recombinant mutant H334Q, phosphorolysis | Corynebacterium callunae | |
0.68 | - |
alpha-D-glucose 1-phosphate | pH 7.0, 30°C, recombinant mutant H334Q, synthesis | Corynebacterium callunae | |
1.27 | - |
alpha-D-glucose 1-phosphate | pH 7.0, 30°C, recombinant mutant H334N, synthesis | Corynebacterium callunae | |
1.5 | - |
starch | pH 7.0, 30°C, recombinant mutant H334N, phosphorolysis | Corynebacterium callunae | |
49 | - |
alpha-D-glucose 1-phosphate | pH 7.0, 30°C, recombinant mutant H334A, synthesis | Corynebacterium callunae | |
52 | - |
alpha-D-glucose 1-phosphate | pH 7.0, 30°C, recombinant wild-type enzyme, synthesis | Corynebacterium callunae | |
74 | - |
starch | pH 7.0, 30°C, recombinant wild-type enzyme, phosphorolysis | Corynebacterium callunae | |
80 | - |
starch | pH 7.0, 30°C, recombinant mutant H334A, phosphorolysis | Corynebacterium callunae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Corynebacterium callunae |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyridoxal 5'-phosphate | - |
Corynebacterium callunae |