Cloned (Comment) | Organism |
---|---|
bgl operon, gene bglB, contruction of a genomic library, DNA and amino acid sequence determination and analysis, determination of bgl operon structure and organization, overexpression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) as His-tagged proteins, subcloning in Escherichia coli strain DH5alpha | Pectobacterium carotovorum |
Protein Variants | Comment | Organism |
---|---|---|
E173A | site-directed mutagenesis, inactive mutant | Pectobacterium carotovorum |
E362A | site-directed mutagenesis, inactive mutant | Pectobacterium carotovorum |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Co2+ | 85% inhibition at 2 mM | Pectobacterium carotovorum | |
Cu2+ | 95% inhibition at 2 mM | Pectobacterium carotovorum | |
Hg2+ | complete inhibition at 2 mM | Pectobacterium carotovorum | |
Mn2+ | 28% inhibition at 2 mM | Pectobacterium carotovorum | |
Zn2+ | 95% inhibition at 2 mM | Pectobacterium carotovorum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.25 | - |
4-nitrophenyl beta-D-glucopyranoside | pH 7.0, 40°C, recombinant enzyme | Pectobacterium carotovorum |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | activates, maximal at 15 mM, 20% activation at 2 mM | Pectobacterium carotovorum | |
additional information | no effect by Ca2+ at 2 mM | Pectobacterium carotovorum |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pectobacterium carotovorum | Q6QGY5 | ssp. carotovorum, strain LY34, bgl operon | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Pectobacterium carotovorum |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
celloheptaose + 6 H2O = 7 beta-D-glucose | residues E173 and E362 are essential for catalytic activity | Pectobacterium carotovorum |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
553 | - |
purified recombinant enzyme, substrate 4-nitrophenyl beta-D-glucopyranoside | Pectobacterium carotovorum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl beta-D-glucopyranoside + H2O | hydrolysis of beta-linkages | Pectobacterium carotovorum | 4-nitrophenol + beta-D-glucose | - |
? |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
- |
Pectobacterium carotovorum |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
20 | 50 | 25% of maximal activity at 20°C and 50°C, half-maximal activity at 28°C and 46°C | Pectobacterium carotovorum |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
loss of 40% of activity within 50 min | Pectobacterium carotovorum |
50 | - |
loss of 90% of activity within 20 min | Pectobacterium carotovorum |
60 | - |
inactivation within 20 min | Pectobacterium carotovorum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
- |
Pectobacterium carotovorum |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
4 | 9 | 20% of maximal activity at pH 5.0 and pH 9.0, half-maximal activity at pH 5.5 and pH 8.5 | Pectobacterium carotovorum |