Cloned (Comment) | Organism |
---|---|
gene cadA, recombinant expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3) | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
F102C/T544C | site-directed mutagenesis, mutant A2 | Escherichia coli |
F14C/K44C | site-directed mutagenesis, mutant B1, the disulfide bond mutation in the decameric interface of wild-type CadA improves its structural stability, and as a result, enhances the pH and thermal stabilities along with organic solvent tolerance, but reduces the catalytic efficiency, compared to the wild-type | Escherichia coli |
F14C/K44C/L7M/N8G | site-directed mutagenesis, the disulfide bond mutation in the decameric interface of wild-type CadA improves its structural stability, and as a result, enhances the pH and thermal stabilities along with organic solvent tolerance compared to the wild-type, addition of mutations L7M and N8G to mutant B1 slightly increases the catalytic efficiency compared to mutant B1 but remains still lower than wild-type | Escherichia coli |
additional information | cadaverine is a major source of many industrial polyamides such as nylon and chelating agents. Cadaverine is produced by the microbial fermentation of glucose to lysine, which is then decarboxylated by lysine decarboxylase CadA. But utilizing CadA for cadaverine production causes enzyme instability. In order to stabilize the CadA homodecamer structure for in vitro decarboxylation reaction, four disulfide bond mutants in the multimeric interfacial region are designed, CadA plasmid library/mutant screening | Escherichia coli |
P233C/L628C | site-directed mutagenesis, mutant C1 | Escherichia coli |
V91C/G445C | site-directed mutagenesis, mutant A1 | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
L-lysine | substrate inhibition at high concentrations | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics | Escherichia coli | |
1.22 | - |
L-lysine | pH 5.6, 45°C, recombinant His6-tagged wild-type enzyme | Escherichia coli | |
1.33 | - |
L-lysine | pH 5.6, 45°C, recombinant His6-tagged mutant F14C/K44C | Escherichia coli | |
1.47 | - |
L-lysine | pH 5.6, 45°C, recombinant His6-tagged mutant F14C/K44C/L7M/N8G | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-lysine | Escherichia coli | - |
cadaverine + CO2 | - |
? | |
L-lysine | Escherichia coli K-12 / B | - |
cadaverine + CO2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0A9H3 | - |
- |
Escherichia coli K-12 / B | P0A9H3 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography and ultrafiltration | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-lysine | - |
Escherichia coli | cadaverine + CO2 | - |
? | |
L-lysine | - |
Escherichia coli K-12 / B | cadaverine + CO2 | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodecamer | a pentamer of homodimers, 10 * 82000, recombinant His6-tagged wild-type enzyme, SDS-PAGE | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
CadA | - |
Escherichia coli |
multimeric lysine decarboxylase | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | 50 | assay at | Escherichia coli |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
80 | - |
purified recombinant His6-tagged wild-type enzyme, pH 5.6, 60% activity remaining | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
146.4 | - |
L-lysine | pH 5.6, 45°C, recombinant His6-tagged wild-type enzyme | Escherichia coli | |
149.3 | - |
L-lysine | pH 5.6, 45°C, recombinant His6-tagged mutant F14C/K44C | Escherichia coli | |
172.1 | - |
L-lysine | pH 5.6, 45°C, recombinant His6-tagged mutant F14C/K44C/L7M/N8G | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5.6 | - |
wild-type enzyme | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyridoxal 5'-phosphate | - |
Escherichia coli |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
112.3 | - |
L-lysine | pH 5.6, 45°C, recombinant His6-tagged mutant F14C/K44C | Escherichia coli | |
117.1 | - |
L-lysine | pH 5.6, 45°C, recombinant His6-tagged mutant F14C/K44C/L7M/N8G | Escherichia coli | |
120.1 | - |
L-lysine | pH 5.6, 45°C, recombinant His6-tagged wild-type enzyme | Escherichia coli |