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Literature summary extracted from

  • Wang, Y.; Li, X.; Chen, X.; Chen, D.
    Directed evolution and characterization of atrazine chlorohydrolase variants with enhanced activity (2013), Biochemistry (Moscow), 78, 1104-1111.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.8.1.8 gene atzA, application of a Haematococcus pluvialis-based method to screen AtzA variants from a random mutagenesis library, genotyping, Haematococcus pluvialis strain H1 is used to test atrazine concentrations, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) in Inclusion bodies, subcloning in Escherichia coli strain DH5alpha-FT Pseudomonas sp.

Protein Variants

EC Number Protein Variants Comment Organism
3.8.1.8 D30G random mutagenesis, the enzyme shows altered kinetics and increased activity compared to the wild-type enzyme Pseudomonas sp.
3.8.1.8 D30G/M315I/R389C/H399Q/N429S/V466A random mutagenesis, the enzyme shows altered kinetics and increased activity compared to the wild-type enzyme Pseudomonas sp.
3.8.1.8 D30G/Q71R/M315I/R389C/H399Q/N429S/V466A random mutagenesis, the enzyme shows altered kinetics and increased activity compared to the wild-type enzyme Pseudomonas sp.
3.8.1.8 F439L random mutagenesis, the enzyme shows altered kinetics and increased activity compared to the wild-type enzyme Pseudomonas sp.
3.8.1.8 H399Q random mutagenesis, the enzyme shows altered kinetics and increased activity compared to the wild-type enzyme Pseudomonas sp.
3.8.1.8 L395P random mutagenesis, the enzyme shows altered kinetics and increased activity compared to the wild-type enzyme Pseudomonas sp.
3.8.1.8 M226V random mutagenesis, the enzyme shows altered kinetics and increased activity compared to the wild-type enzyme Pseudomonas sp.
3.8.1.8 M226V/V278A random mutagenesis, the enzyme shows altered kinetics and increased activity compared to the wild-type enzyme Pseudomonas sp.
3.8.1.8 M315I random mutagenesis, the enzyme shows altered kinetics and increased activity compared to the wild-type enzyme Pseudomonas sp.
3.8.1.8 M315I/H399Q/N429S/V466A random mutagenesis, the enzyme shows altered kinetics and increased activity compared to the wild-type enzyme Pseudomonas sp.
3.8.1.8 M337T random mutagenesis, the enzyme shows altered kinetics and increased activity compared to the wild-type enzyme Pseudomonas sp.
3.8.1.8 N429S random mutagenesis, the enzyme shows altered kinetics and increased activity compared to the wild-type enzyme Pseudomonas sp.
3.8.1.8 Q71R random mutagenesis, the enzyme shows altered kinetics and increased activity compared to the wild-type enzyme Pseudomonas sp.
3.8.1.8 R389S random mutagenesis, the enzyme shows altered kinetics and increased activity compared to the wild-type enzyme Pseudomonas sp.
3.8.1.8 T195A random mutagenesis, the enzyme shows altered kinetics and increased activity compared to the wild-type enzyme Pseudomonas sp.
3.8.1.8 T195A/M337T/F439L random mutagenesis, the enzyme shows altered kinetics and increased activity compared to the wild-type enzyme Pseudomonas sp.
3.8.1.8 V12A random mutagenesis, the enzyme shows altered kinetics and increased activity compared to the wild-type enzyme Pseudomonas sp.
3.8.1.8 V278A random mutagenesis, the enzyme shows altered kinetics and increased activity compared to the wild-type enzyme Pseudomonas sp.
3.8.1.8 V466A random mutagenesis, the enzyme shows altered kinetics and increased activity compared to the wild-type enzyme Pseudomonas sp.
3.8.1.8 V58A/H80R/T121A random mutagenesis, the enzyme shows altered kinetics and increased activity compared to the wild-type enzyme Pseudomonas sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.8.1.8 additional information
-
 additional information kinetic parameters of diverse mutants relative to the wild-type values, overview Pseudomonas sp.
3.8.1.8 0.146
-
 Atrazine recombinant wild-type enzyme, pH 7.2, 25°C Pseudomonas sp.

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.8.1.8 atrazine + H2O Pseudomonas sp.
-
 4-(ethylamino)-2-hydroxy-6-(isopropylamino)-1,3,5-triazine + chloride
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.8.1.8 Pseudomonas sp. P72156 gene atzA
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.8.1.8 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) solubilized from inclusion bodies by nickel affinity chromatography Pseudomonas sp.

Renatured (Commentary)

EC Number Renatured (Comment) Organism
3.8.1.8 solubilization of recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) inclusion bodies Pseudomonas sp.

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.8.1.8 2.6
-
 purified recombinant wild-type enzyme, 0.046 mM atrazine, pH 7.2, 25°C Pseudomonas sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.8.1.8 atrazine + H2O
-
 Pseudomonas sp. 4-(ethylamino)-2-hydroxy-6-(isopropylamino)-1,3,5-triazine + chloride
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.8.1.8 AtzA
-
 Pseudomonas sp.

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.8.1.8 25
-
 assay at Pseudomonas sp.

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.8.1.8 1.6
-
 Atrazine recombinant wild-type enzyme, pH 7.2, 25°C Pseudomonas sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.8.1.8 7.2
-
 assay at Pseudomonas sp.

General Information

EC Number General Information Comment Organism
3.8.1.8 additional information homology modeling of enzyme AtzA, structure modeling and distribution of substitution sites Pseudomonas sp.

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.8.1.8 11
-
 Atrazine recombinant wild-type enzyme, pH 7.2, 25°C Pseudomonas sp.