Application | Comment | Organism |
---|---|---|
biotechnology | bioremediation | Escherichia coli |
Cloned (Comment) | Organism |
---|---|
five amino acids within the substrate-binding pocket of AtzA are chosen for randomization yields one variant enzyme with 20fold-greater catalytic efficiency than that of wild-type AtzA | Thermotoga maritima |
Protein Variants | Comment | Organism |
---|---|---|
A216A/T217D/T219E/A220A/D250D | site-directed mutagenesis, AtzA-variant 288. Km 49 microM, kcat 4.3 s-1, kcat/Km 88000 s-1*M-1 | Escherichia coli |
A216A/T217S/T219H/A220G/D250Y | site-directed mutagenesis, AtzA-variant 431. Km 76 microM, kcat 8.0 s-1, kcat/Km 110000 s-1*M-1 | Escherichia coli |
A216G/T217D/T219A/A220V/D250W | site-directed mutagenesis, AtzA-variant 305. Km 95 microM, kcat 6.2 s-1, kcat/Km 65000 s-1*M-1 | Escherichia coli |
A216G/T217D/T219A/A220V/D250W | AtzA mutant | Thermotoga maritima |
A216G/T217D/T219G/A220H/D250D | site-directed mutagenesis, AtzA-variant 734. Km 62 microM, kcat 15.1 s-1, kcat/Km 240000 s-1*M-1 | Escherichia coli |
A216G/T217D/T219G/A220H/D250D | AtzA mutant | Thermotoga maritima |
A216G/T217D/T219G/A220H/D250G | site-directed mutagenesis, AtzA-variant 430. Km 90 microM, kcat 12.7 s-1, kcat/Km 140000 s-1*M-1 | Escherichia coli |
A216G/T217D/T219G/A220H/D250G | AtzA mutant | Thermotoga maritima |
A216H/T217A/T219E/A220S/D250S | site-directed mutagenesis, AtzA-variant 662. Km 100 microM, kcat 7.4 s-1, kcat/Km 74000 s-1*M-1 | Escherichia coli |
A216H/T217A/T219E/A220S/D250S | AtzA mutant | Thermotoga maritima |
A216S/T217A/T219P/A220F/D250G | site-directed mutagenesis, AtzA-variant 357. Km 67 microM, kcat 2.3 s-1, kcat/Km 34000 s-1*M-1 | Escherichia coli |
A216S/T217A/T219P/A220F/D250G | AtzA mutant | Thermotoga maritima |
A216S/T217D/T219G/A220S | AtzA mutant | Thermotoga maritima |
A216S/T217D/T219G/A220S/D250D | site-directed mutagenesis, AtzA-variant 841. Km 105 microM, kcat 8.5 s-1, kcat/Km 81000 s-1*M-1 | Escherichia coli |
A216S/T217D/T219V/A220H/D250G | site-directed mutagenesis, AtzA-variant 297. Km 92 microM, kcat 6.8 s-1, kcat/Km 74000 s-1*M-1 | Escherichia coli |
A216S/T217D/T219V/A220H/D250G | AtzA mutant | Thermotoga maritima |
A216Y/T217D/A220H/D250E | AtzA mutant | Thermotoga maritima |
A216Y/T217D/T219Y/A220H/D250V | site-directed mutagenesis, AtzA-variant 422. Km 92 microM, kcat 6.5 s-1, kcat/Km 71000 s-1*M-1 | Escherichia coli |
A216Y/T217D/T219Y/A220H/D250V | AtzA mutant | Thermotoga maritima |
additional information | A216Y/T217D/T219T/A220H/D250E, site-directed mutagenesis, AtzA best variant consensus | Escherichia coli |
T217D/T219E | AtzA mutant | Thermotoga maritima |
T217S/T219H/A220G/D250Y | AtzA mutant | Thermotoga maritima |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | reduction in Km observed in the AtzA variants has allowed the full kinetic profile for the AtzA-catalyzed dechlorination of atrazine to be determined for the first time, revealing the hitherto-unreported substrate cooperativity in AtzA | Escherichia coli | |
0.049 | - |
Atrazine | A216A/T217D/T219E/A220A/D250D, site-directed mutagenesis, AtzA-variant 288, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
0.049 | - |
Atrazine | T217D/T219E, AtzA mutant, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima | |
0.062 | - |
Atrazine | A216G/T217D/T219G/A220H/D250D, site-directed mutagenesis, AtzA-variant 734, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
0.062 | - |
Atrazine | A216G/T217D/T219G/A220H/D250D, AtzA mutant, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima | |
0.067 | - |
Atrazine | A216S/T217A/T219P/A220F/D250G, site-directed mutagenesis, AtzA-variant 357, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
0.067 | - |
Atrazine | A216S/T217A/T219P/A220F/D250G, AtzA mutant, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima | |
0.076 | - |
Atrazine | A216A/T217S/T219H/A220G/D250Y, site-directed mutagenesis, AtzA-variant 431, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
0.076 | - |
Atrazine | T217S/T219H/A220G/D250Y, AtzA mutant, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima | |
0.09 | - |
Atrazine | A216G/T217D/T219G/A220H/D250G, site-directed mutagenesis, AtzA-variant 430, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
0.092 | - |
Atrazine | A216S/T217D/T219V/A220H/D250G, site-directed mutagenesis, AtzA-variant 297, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
0.092 | - |
Atrazine | A216Y/T217D/T219Y/A220H/D250V, site-directed mutagenesis, AtzA-variant 422, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
0.092 | - |
Atrazine | A216S/T217D/T219V/A220H/D250G, AtzA mutant, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima | |
0.092 | - |
Atrazine | A216Y/T217D/T219Y/A220H/D250V, AtzA mutant, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima | |
0.093 | - |
Atrazine | A216Y/T217D/T219T/A220H/D250E, site-directed mutagenesis, AtzA best variant consensus, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
0.093 | - |
Atrazine | A216Y/T217D/A220H/D250E, AtzA mutant, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima | |
0.095 | - |
Atrazine | A216G/T217D/T219A/A220V/D250W, site-directed mutagenesis, AtzA-variant 305, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
0.095 | - |
Atrazine | A216G/T217D/T219A/A220V/D250W, AtzA mutant, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima | |
0.1 | - |
Atrazine | A216H/T217A/T219E/A220S/D250S, site-directed mutagenesis, AtzA-variant 662, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
0.1 | - |
Atrazine | A216H/T217A/T219E/A220S/D250S, AtzA mutant, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima | |
0.105 | - |
Atrazine | A216S/T217D/T219G/A220S/D250D, site-directed mutagenesis, AtzA-variant 841, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
0.105 | - |
Atrazine | A216S/T217D/T219G/A220S, AtzA mutant, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima | |
0.153 | - |
Atrazine | more than 0.153 mM. AtzA wild type, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
0.153 | - |
Atrazine | AtzA wild-type, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima | |
0.9 | - |
Atrazine | A216G/T217D/T219G/A220H/D250G, AtzA mutant, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
atrazine + H2O | Escherichia coli | 6-chloro-N2-ethyl-N4-isopropyl-1,3,5-triazine-2,4-diamine, herbicide | 4-(ethylamino)-2-hydroxy-6-(isopropylamino)-1,3,5-triazine + HCl | - |
? | |
atrazine + H2O | Escherichia coli JM109 | 6-chloro-N2-ethyl-N4-isopropyl-1,3,5-triazine-2,4-diamine, herbicide | 4-(ethylamino)-2-hydroxy-6-(isopropylamino)-1,3,5-triazine + HCl | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
JM109 | - |
Escherichia coli JM109 | - |
JM109 | - |
Thermotoga maritima | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
atrazine + H2O | - |
Thermotoga maritima | 4-(ethylamino)-2-hydroxy-6-(isopropylamino)-1,3,5-triazine + HCl | - |
? | |
atrazine + H2O | 6-chloro-N2-ethyl-N4-isopropyl-1,3,5-triazine-2,4-diamine, herbicide | Escherichia coli | 4-(ethylamino)-2-hydroxy-6-(isopropylamino)-1,3,5-triazine + HCl | - |
? | |
atrazine + H2O | 6-chloro-N2-ethyl-N4-isopropyl-1,3,5-triazine-2,4-diamine, herbicide | Escherichia coli JM109 | 4-(ethylamino)-2-hydroxy-6-(isopropylamino)-1,3,5-triazine + HCl | - |
? |
Synonyms | Comment | Organism |
---|---|---|
atrazine chlorohydrolase | - |
Escherichia coli |
atrazine chlorohydrolase | - |
Thermotoga maritima |
AtzA | - |
Escherichia coli |
AtzA | - |
Thermotoga maritima |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.2 | - |
Atrazine | more than 0.153 mM. AtzA wild type, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
2.2 | - |
Atrazine | AtzA wild-type, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima | |
2.3 | - |
Atrazine | A216S/T217A/T219P/A220F/D250G, site-directed mutagenesis, AtzA-variant 357, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
2.3 | - |
Atrazine | A216S/T217A/T219P/A220F/D250G, AtzA mutant, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima | |
4.3 | - |
Atrazine | A216A/T217D/T219E/A220A/D250D, site-directed mutagenesis, AtzA-variant 288, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
4.3 | - |
Atrazine | T217D/T219E, AtzA mutant, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima | |
6.2 | - |
Atrazine | A216G/T217D/T219A/A220V/D250W, site-directed mutagenesis, AtzA-variant 305, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
6.2 | - |
Atrazine | A216G/T217D/T219A/A220V/D250W, AtzA mutant, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima | |
6.5 | - |
Atrazine | A216Y/T217D/T219Y/A220H/D250V, site-directed mutagenesis, AtzA-variant 422, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
6.5 | - |
Atrazine | A216Y/T217D/T219Y/A220H/D250V, AtzA mutant, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima | |
6.8 | - |
Atrazine | A216S/T217D/T219V/A220H/D250G, site-directed mutagenesis, AtzA-variant 297, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
6.8 | - |
Atrazine | A216S/T217D/T219V/A220H/D250G, AtzA mutant, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima | |
7.4 | - |
Atrazine | A216H/T217A/T219E/A220S/D250S, site-directed mutagenesis, AtzA-variant 662, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
7.4 | - |
Atrazine | A216H/T217A/T219E/A220S/D250S, AtzA mutant, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima | |
8 | - |
Atrazine | A216A/T217S/T219H/A220G/D250Y, site-directed mutagenesis, AtzA-variant 431, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
8 | - |
Atrazine | T217S/T219H/A220G/D250Y, AtzA mutant, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima | |
8.5 | - |
Atrazine | A216S/T217D/T219G/A220S/D250D, site-directed mutagenesis, AtzA-variant 841, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
8.5 | - |
Atrazine | A216S/T217D/T219G/A220S, AtzA mutant, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima | |
12.7 | - |
Atrazine | A216G/T217D/T219G/A220H/D250G, site-directed mutagenesis, AtzA-variant 430, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
12.7 | - |
Atrazine | A216G/T217D/T219G/A220H/D250G, AtzA mutant, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima | |
15.1 | - |
Atrazine | A216G/T217D/T219G/A220H/D250D, site-directed mutagenesis, AtzA-variant 734, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
15.1 | - |
Atrazine | A216G/T217D/T219G/A220H/D250D, AtzA mutant, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima | |
27.9 | - |
Atrazine | A216Y/T217D/T219T/A220H/D250E, site-directed mutagenesis, AtzA best variant consensus, kinetic data obtained using 4.6 to 153 microM atrazine and 10 nM enzyme | Escherichia coli | |
27.9 | - |
Atrazine | A216Y/T217D/A220H/D250E, AtzA mutant, substrate concentration 153 microM, enzyme concentration 10 nM, measuring absorbance of atrazine after reaction with pyridine at 436 nm | Thermotoga maritima |