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Literature summary for 3.8.1.8 extracted from

  • Scott, C.; Jackson, C.J.; Coppin, C.W.; Mourant, R.G.; Hilton, M.E.; Russell, R.J.; Oakeshott, J.G.
    Atrazine chlorohydrolase: catalytic improvement and evolution (2009), Appl. Environ. Microbiol., 75, 2184-2191.
    View publication on PubMedView publication on EuropePMC

Application

Application Comment Organism
biotechnology bioremediation Escherichia coli

Cloned(Commentary)

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

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 [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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Escherichia coli
-
JM109
-
Escherichia coli JM109
-
JM109
-
Thermotoga maritima
-
-
-

Substrates and Products (Substrate)

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

Synonyms Comment Organism
atrazine chlorohydrolase
-
Escherichia coli
atrazine chlorohydrolase
-
Thermotoga maritima
AtzA
-
Escherichia coli
AtzA
-
Thermotoga maritima

Turnover Number [1/s]

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