Cloned (Comment) | Organism |
---|---|
gene gadB, sequence comparisons, recombinant expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3). The expression level of both truncated mutants in Escherichia coli at 25°C is much higher than that at 37°C | Lactiplantibacillus plantarum subsp. plantarum |
Protein Variants | Comment | Organism |
---|---|---|
additional information | generation of C-terminally truncated (DELTA3 and DELTA11 residues) mutants, their enzyme activities are compared with that of the wild-type enzyme at different pH values. Unlike the wild-type GAD, the mutants show pronounced catalytic activity in a broad pH range of 4.0-8.0, suggesting that the C-terminal region is involved in the pH dependence of GAD activity | Lactiplantibacillus plantarum subsp. plantarum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics | Lactiplantibacillus plantarum subsp. plantarum | |
22.8 | - |
L-glutamate | pH 5.0, 37°C, recombinant wild-type enzyme | Lactiplantibacillus plantarum subsp. plantarum | |
33.8 | - |
L-glutamate | pH 5.0, 37°C, recombinant enzyme mutant DELTA11 | Lactiplantibacillus plantarum subsp. plantarum | |
122.1 | - |
L-glutamate | pH 7.0, 37°C, recombinant wild-type enzyme | Lactiplantibacillus plantarum subsp. plantarum | |
137.1 | - |
L-glutamate | pH 7.0, 37°C, recombinant enzyme mutant DELTA11 | Lactiplantibacillus plantarum subsp. plantarum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-glutamate | Lactiplantibacillus plantarum subsp. plantarum | - |
4-aminobutanoate + CO2 | - |
? | |
L-glutamate | Lactiplantibacillus plantarum subsp. plantarum ATCC 14917 / JCM 1149 / CGMCC 1.2437 | - |
4-aminobutanoate + CO2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Lactiplantibacillus plantarum subsp. plantarum | D7VEX3 | - |
- |
Lactiplantibacillus plantarum subsp. plantarum ATCC 14917 / JCM 1149 / CGMCC 1.2437 | D7VEX3 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, tag cleavage through thrombin, and dialysis, followed by gel filtration | Lactiplantibacillus plantarum subsp. plantarum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-glutamate | - |
Lactiplantibacillus plantarum subsp. plantarum | 4-aminobutanoate + CO2 | - |
? | |
L-glutamate | - |
Lactiplantibacillus plantarum subsp. plantarum ATCC 14917 / JCM 1149 / CGMCC 1.2437 | 4-aminobutanoate + CO2 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
GAD | - |
Lactiplantibacillus plantarum subsp. plantarum |
GadB | - |
Lactiplantibacillus plantarum subsp. plantarum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
- |
Lactiplantibacillus plantarum subsp. plantarum |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | 60 | over 60% activity within this range | Lactiplantibacillus plantarum subsp. plantarum |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
70 | 90 | the recombinant enzyme with a thermal half-life of 46 min at 90°C retains approximately 70% of initial activity after incubation at 70°C for 9 h, whereas the enzyme is rapidly inactivated at 100°C | Lactiplantibacillus plantarum subsp. plantarum |
90 | - |
irreversible thermal unfolding of the purified recombinant detagged enzyme | Lactiplantibacillus plantarum subsp. plantarum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.015 | - |
L-glutamate | pH 7.0, 37°C, recombinant wild-type enzyme | Lactiplantibacillus plantarum subsp. plantarum | |
0.485 | - |
L-glutamate | pH 7.0, 37°C, recombinant enzyme mutant DELTA11 | Lactiplantibacillus plantarum subsp. plantarum | |
21.75 | - |
L-glutamate | pH 5.0, 37°C, recombinant wild-type enzyme | Lactiplantibacillus plantarum subsp. plantarum | |
40.25 | - |
L-glutamate | pH 5.0, 37°C, recombinant enzyme mutant DELTA11 | Lactiplantibacillus plantarum subsp. plantarum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5 | - |
- |
Lactiplantibacillus plantarum subsp. plantarum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyridoxal 5'-phosphate | - |
Lactiplantibacillus plantarum subsp. plantarum |
General Information | Comment | Organism |
---|---|---|
additional information | the GAD C-terminal region (Ile454-Thr468) plays an important role in the pH dependence of catalysis. Homology modeling of GAD | Lactiplantibacillus plantarum subsp. plantarum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00012 | - |
L-glutamate | pH 7.0, 37°C, recombinant wild-type enzyme | Lactiplantibacillus plantarum subsp. plantarum | |
0.0035 | - |
L-glutamate | pH 7.0, 37°C, recombinant enzyme mutant DELTA11 | Lactiplantibacillus plantarum subsp. plantarum | |
0.954 | - |
L-glutamate | pH 5.0, 37°C, recombinant wild-type enzyme | Lactiplantibacillus plantarum subsp. plantarum | |
1.19 | - |
L-glutamate | pH 5.0, 37°C, recombinant enzyme mutant DELTA11 | Lactiplantibacillus plantarum subsp. plantarum |