Application | Comment | Organism |
---|---|---|
synthesis | the enzyme can be used for industrial production of cadaverine, especially mutant T88S is a promising biocatalyst | Escherichia coli |
Cloned (Comment) | Organism |
---|---|
gene cadA, recombinant expression of His-tagged wild-type and T88 mutants in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
additional information | engineering the decameric interface for potential for industrial applications | Escherichia coli |
T88D | site-directed mutagenesis, the mutant shows decreased thermostability compared to the wild-type enzyme | Escherichia coli |
T88F | site-directed mutagenesis, the mutant shows increased thermostability compared to the wild-type enzyme | Escherichia coli |
T88N | site-directed mutagenesis, the mutant is expressed in inclusion bodies and shows no clear activity | Escherichia coli |
T88P | site-directed mutagenesis, the mutant is expressed in inclusion bodies and shows no clear activity | Escherichia coli |
T88Q | site-directed mutagenesis, the mutant is expressed in inclusion bodies and shows no clear activity | Escherichia coli |
T88S | site-directed mutagenesis, the mutant shows higher thermostability with a 2.9fold increase in the half-life at 70°C (from 11 min to 32 min) and increased melting temperature (from 76°C to 78°C). The specific activity and pH stability of T88S at pH 8.0 are increased to 164 U/mg and 78% compared to 58 U/mg and 57% for the wild-type enzyme. The productivity of cadaverine with T88S is 40 g/l/h in contrast to 28 g/l/h with wild-type enzyme. The mutant is a promising biocatalyst for industrial production of cadaverine. No additional hydrogen bond is formed when T88 is substituted by D, F, or S, and the improved stability may be attributed to the favorable atom and torsion angle potentials | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-lysine | Escherichia coli | - |
cadaverine + CO2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0A9H3 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant T88S 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 | - |
? |
Subunits | Comment | Organism |
---|---|---|
decamer | 10 * 81000, recombinant His-tagged wild-type and mutant T88S, SDS-PAGE | Escherichia coli |
More | three-dimensional structure analysis of enzyme CadA, overview | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
CadA | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
recombinant His-tagged wild-type enzyme | Escherichia coli |
55 | - |
recombinant His-tagged mutant T88S | Escherichia coli |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
purified wild-type CadA, residual activity is 86.6% after 12 h | Escherichia coli |
70 | - |
crude enzymes, half life for the wild-type enzyme is 11 min, the half-lives of T88 are: 21-32 min for T88S, 7 min for T88D, 12 min for T88F. after 50 min at 70°C, the wild-type enzyme is inactivated, while mutant T88S still shows 23.1% activity | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5.5 | - |
recombinant His-tagged wild-type enzyme | Escherichia coli |
7 | - |
recombinant His-tagged mutant T88S | Escherichia coli |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
5 | - |
10 h at 37°C, about 80% activity remaining for the wild-type enzyme, and about 75% activity remaining for the recombinant His-tagged mutant T88S | Escherichia coli |
6 | - |
recombinant His-tagged wild-type enzyme, most stable at pH 6.0, over 90% activity remaining after 10 h at 37°C, about 85% activity remaining for the mutant T88S | Escherichia coli |
7 | - |
recombinant His-tagged mutant T88S, most stable at pH 7.0, over 90% activity remaining after 10 h at 37°C, about 60% activity remaining for the wild-type enzyme | Escherichia coli |
8.5 | - |
10 h at 37°C, about 40% activity remaining for the wild-type enzyme, and about 70% activity remaining for the recombinant His-tagged mutant T88S | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyridoxal 5'-phosphate | - |
Escherichia coli |