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Literature summary for 3.8.1.8 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
Search for more literature for 3.8.1.8

Cloned(Commentary)

Cloned (Comment) Organism
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

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

KM Value [mM]

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Pseudomonas sp. P72156 gene atzA
-

Purification (Commentary)

Purification (Comment) Organism
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)

Renatured (Comment) Organism
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]

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

Substrates and Products (Substrate)

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

Synonyms

Synonyms Comment Organism
AtzA
-
 Pseudomonas sp.

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

pH Optimum

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

General Information

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

kcat/KM [mM/s]

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