Cloned (Comment) | Organism |
---|---|
gene gad, DNA and amino acid sequence determination and analysis, sequence comparisons, functional recombinant expression of soluble His-tagged enzyme in Escherichia coli strain BL21(DE3) | Saccharomyces cerevisiae |
Protein Variants | Comment | Organism |
---|---|---|
additional information | optimization of bioproduction of gamma-GABA using recombinant Escherichia coli expressing a GAD enzyme derived from eukaryote. The optimal medium for Escherichia coli-ScGAD cultivation and expression is 10 g/l lactose, 5 g/l glycerol, 20 g/l yeast extract, and 10 g/l sodium chloride, resulting in an activity of 55 U/ml medium, three times higher than that of using Luria-Bertani (LB) medium. The maximal concentration of gamma-GABA is 245 g/l whereas L-glutamic acid is near completely converted | Saccharomyces cerevisiae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
11.4 | - |
L-glutamate | pH 4.2, 37°C, recombinant enzyme | Saccharomyces cerevisiae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-glutamate | Saccharomyces cerevisiae | - |
4-aminobutanoate + CO2 | - |
? | |
L-glutamate | Saccharomyces cerevisiae NX-1 | - |
4-aminobutanoate + CO2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharomyces cerevisiae | - |
- |
- |
Saccharomyces cerevisiae NX-1 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and ultrafiltration, to homogeneity | Saccharomyces cerevisiae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-glutamate | - |
Saccharomyces cerevisiae | 4-aminobutanoate + CO2 | - |
? | |
L-glutamate | - |
Saccharomyces cerevisiae NX-1 | 4-aminobutanoate + CO2 | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 66000, recombinant His-tagged enzyme, SDS-PAGE, x * 65986, sequence calculation | Saccharomyces cerevisiae |
Synonyms | Comment | Organism |
---|---|---|
GAD | - |
Saccharomyces cerevisiae |
ScGAD | - |
Saccharomyces cerevisiae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
- |
Saccharomyces cerevisiae |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.537 | - |
L-glutamate | pH 4.2, 37°C, recombinant enzyme | Saccharomyces cerevisiae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
4.2 | - |
- |
Saccharomyces cerevisiae |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | the cofactor of ScGAD is verified to be either pyridoxal 5'-phosphate (PLP) or pyridoxal. The optimal concentration of either cofactor is 50 mg/l. GAD requires a coenzyme for activity while inactive apo-GAD forms a GAD-PLP complex (holo-GAD) to obtain its activity | Saccharomyces cerevisiae | |
pyridoxal 5'-phosphate | - |
Saccharomyces cerevisiae |