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Literature summary extracted from

  • Seffernick, J.L.; Aleem, A.; Osborne, J.P.; Johnson, G.; Sadowsky, M.J.; Wackett, L.P.
    Hydroxyatrazine N-ethylaminohydrolase (AtzB): an amidohydrolase superfamily enzyme catalyzing deamination and dechlorination (2007), J. Bacteriol., 189, 6989-6997.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.4.43 gene atzB, expression in Escherichia coli strain DH5alpha Pseudomonas sp.
3.8.1.8 using the vectors pACYC184 and pMD4 for expression in Escherichia coli DH5alpha cells Pseudomonas sp.

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.4.43 1,10-phenanthroline 72% inhibition, not due to metal removal Pseudomonas sp.
3.5.4.43 8-Hydroxyquinoline-5-sulfonic acid
-
 Pseudomonas sp.
3.5.4.43 Chelex-100 resin
-
 Pseudomonas sp.
3.5.4.43 dipicolinic acid
-
 Pseudomonas sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.8.1.8 0.02
-
 hydroxyatrazine
-
 Pseudomonas sp.
3.8.1.8 0.02
-
 2-(N-ethylamino)-4-hydroxy-6-(N-isopropylamino)-1,3,5-triazine deamination Pseudomonas sp.
3.8.1.8 0.029
-
 2,4-diisopropylamino-6-hydroxy-1,3,5-triazine deamination Pseudomonas sp.
3.8.1.8 0.04
-
 6-hydroxy-4-(N-isopropylamino)-2-(N-methylamino)-1,3,5-triazine deamination Pseudomonas sp.
3.8.1.8 0.12
-
 2-chloro-4-hydroxy-6-(N-isopropylamino)-1,3,5-triazine dechlorination Pseudomonas sp.
3.8.1.8 0.23
-
 2-chloro-4-(N-ethylamino)-6-hydroxy-1,3,5-triazine dechlorination Pseudomonas sp.

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.8.1.8 Zn2+
-
 Pseudomonas sp.

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.5.4.43 52000
-
 2 * 52000, recombinant AtzB, SDS-PAGE Pseudomonas sp.
3.5.4.43 105000
-
 recombinant AtzB, gel filtration Pseudomonas sp.
3.8.1.8 52000
-
 subunit molecular mass, determind by SDS-Page Pseudomonas sp.
3.8.1.8 105000
-
 holoenzyme molecular mass, determind by gel-filtration 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 hydroxyatrazine + H2O Pseudomonas sp.
-
 N-isopropylammelide + ethylamine
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.5.4.43 Pseudomonas sp.
-
 gene atzB
-
3.8.1.8 Pseudomonas sp.
-
 Pseudomonas sp. starin ADP
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.4.43 recombinant enzyme 6.6fold from Escherichia coli strain DH5alpha to homogeneity by ammonium sulfate fractionation, hydroxyapatite chromatography, and gel filtration Pseudomonas sp.
3.8.1.8 using a CHT ceramic hydroxyapatite type I column Pseudomonas sp.

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.5.4.43 1.05
-
 purified recombinant enzyme Pseudomonas sp.
3.8.1.8 0.16
-
 crude extract Pseudomonas sp.
3.8.1.8 0.33
-
 prurification step ammonium sulfate precipitation Pseudomonas sp.
3.8.1.8 1.05
-
 prurification step ceramic hydroxyapatite type I column Pseudomonas sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.4.43 2,4-di(N-isobutylamino)-6-hydroxy-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2,4-di(N-sec-butylamino)-6-hydroxy-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2,4-di(N-tert-butylamino)-6-hydroxy-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2,4-diamino-6-hydroxy-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2,4-dichloro-6-amino-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2,4-diethylamino-6-hydroxy-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2,4-diisopropylamino-6-hydroxy-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2,4-dimethylamino-6-hydroxy-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2-(N-ethyl-N-methylamino)-4-ethylamino-6-hydroxy-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2-(N-ethylamino)-4-hydroxy-6-(N-hydroxyethylamino)-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2-(N-ethylamino)-4-hydroxy-6-(N-isopropylamino)-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2-amino-4-(N-ethylamino)-6-(N-isopropylamino)-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2-amino-4-chloro-6-hydroxy-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2-amino-4-hydroxy-6-(N-hydroxyethylamino)-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2-amino-4-hydroxy-6-(N-isopropylamino)-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2-amino-6-(N-ethylamino)-4-hydroxy-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2-azido-4-(N-ethylamino)-6-(N-isopropylamino)-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2-chloro-4-(N-ethylamino)-6-hydroxy-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2-chloro-4-amino-6-hydroxy-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2-chloro-4-hydroxy-6-(N-isopropylamino)-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2-cyano-4-(N-ethylamino)-6-(N-isopropylamino)-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2-fluoro-4-(N-ethylamino)-6-(N-isopropylamino)-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2-hydroxy-4,6-di(N-hydroxyethylamino)-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2-hydroxy-4-(N-ethylamino)-6-(N-isopropylamino)-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2-hydroxy-4-(N-isopropylamino)-6-(N-(3-methoxypropyl)amino)-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2-isopropylamino-4-ethylamino-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 2-mercapto-4-isopropylamino-6-ethylamino-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 4-(N-ethylamino)-6-hydroxy-2-(N-methylamino)-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 6-hydroxy-4-(N-isopropylamino)-2-(N-methylamino)-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.5.4.43 additional information the enzyme performs deamination and dechlorination, substrate specificity, overview Pseudomonas sp. ?
-
 ?
3.8.1.8 2,4-di(N-isobutylamino)-6-hydroxy-1,3,5-triazine + H2O
-
 Pseudomonas sp. 6-[(2-methylpropyl)amino]-1,3,5-triazine-2,4-diol + isobutylamine
-
 ?
3.8.1.8 2,4-di(N-secbutylamino)-6-hydroxy-1,3,5-triazine + H2O
-
 Pseudomonas sp. 6-[(1-methylpropyl)amino]-1,3,5-triazine-2,4-diol + butan-2-amine
-
 ?
3.8.1.8 2,4-di(N-tertbutylamino)-6-hydroxy-1,3,5-triazine + H2O
-
 Pseudomonas sp. 6-(tert-butylamino)-1,3,5-triazine-2,4-diol + tert-butylamine
-
 ?
3.8.1.8 2,4-diamino-6-hydroxy-1,3,5-triazine + H2O
-
 Pseudomonas sp. 6-amino-1,3,5-triazine-2,4-diol + NH3
-
 ?
3.8.1.8 2,4-diethylamino-6-hydroxy-1,3,5-triazine + H2O
-
 Pseudomonas sp. 6-(methylamino)-1,3,5-triazine-2,4-diol + ethylamine
-
 ?
3.8.1.8 2,4-diisopropylamino-6-hydroxy-1,3,5-triazine + H2O
-
 Pseudomonas sp. 6-[(1-methylethyl)amino]-1,3,5-triazine-2,4-diol + propan-2-amine
-
 ?
3.8.1.8 2,4-dimethylamino-6-hydroxy-1,3,5-triazine + H2O
-
 Pseudomonas sp. 6-(methylamino)-1,3,5-triazine-2,4-diol + methylamine
-
 ?
3.8.1.8 2-(N-ethyl-N-methylamino)-4-ethylamino-6-hydroxy-1,3,5-triazine + H2O
-
 Pseudomonas sp. 6-[ethyl(methyl)amino]-1,3,5-triazine-2,4-diol + ethylamine
-
 ?
3.8.1.8 2-(N-ethylamino)-4-hydroxy-6-(N-hydroxyethylamino)-1,3,5-triazine + H2O
-
 Pseudomonas sp. 6-[(2-hydroxyethyl)amino]-1,3,5-triazine-2,4-diol + ethylamine
-
 ?
3.8.1.8 2-(N-ethylamino)-4-hydroxy-6-(N-isopropylamino)-1,3,5-triazine + H2O
-
 Pseudomonas sp. 6-(ethylamino)-1,3,5-triazine-2,4-diol + propan-2-amine
-
 ?
3.8.1.8 2-amino-4-chloro-6-hydroxy-1,3,5-triazine + H2O
-
 Pseudomonas sp. 6-amino-1,3,5-triazine-2,4-diol + Cl-
-
 ?
3.8.1.8 2-amino-4-hydroxy-6-(N-hydroxyethylamino)-1,3,5-triazine + H2O
-
 Pseudomonas sp. 6-[(2-hydroxyethyl)amino]-1,3,5-triazine-2,4-diol + NH3
-
 ?
3.8.1.8 2-amino-4-hydroxy-6-(N-isopropylamino)-1,3,5-triazine + H2O
-
 Pseudomonas sp. 6-(isopropylamino)-1,3,5-triazine-2,4-diol + NH3
-
 ?
3.8.1.8 2-amino-6-(N-ethylamino)-4-hydroxy-1,3,5-triazine + H2O
-
 Pseudomonas sp. 6-(ethylamino)-1,3,5-triazine-2,4-diol + NH3
-
 ?
3.8.1.8 2-chloro-4-(N-ethylamino)-6-hydroxy-1,3,5-triazine + H2O
-
 Pseudomonas sp. 6-(ethylamino)-1,3,5-triazine-2,4-diol + Cl-
-
 ?
3.8.1.8 2-chloro-4-hydroxy-6-(N-isopropylamino)-1,3,5-triazine + H2O
-
 Pseudomonas sp. 6-(isopropylamino)-1,3,5-triazine-2,4-diol + Cl-
-
 ?
3.8.1.8 2-hydroxy-4,6-di(N-hydroxyethylamino)-1,3,5-triazine + H2O
-
 Pseudomonas sp. 6-[(2-hydroxyethyl)amino]-1,3,5-triazine-2,4-diol + 2-aminoethanol
-
 ?
3.8.1.8 2-hydroxy-4-(N-isopropylamino)-6-(N-(3-methoxypropyl)amino)-1,3,5-triazine + H2O
-
 Pseudomonas sp. ?
-
 ?
3.8.1.8 6-(ethylamino)-4-(methylamino)-1,3,5-triazin-2-ol + H2O
-
 Pseudomonas sp. 6-(ethylamino)-1,3,5-triazine-2,4-diol + methylamine
-
 ?
3.8.1.8 6-hydroxy-4-(N-isopropylamino)-2-(N-methylamino)-1,3,5-triazine + H2O
-
 Pseudomonas sp. 6-[(1-methylethyl)amino]-1,3,5-triazine-2,4-diol + methylamine
-
 ?
3.8.1.8 hydroxyatrazine + H2O
-
 Pseudomonas sp. N-isopropylammelide + ethylamine
-
 ?

Subunits

EC Number Subunits Comment Organism
3.5.4.43 dimer 2 * 52000, recombinant AtzB, SDS-PAGE Pseudomonas sp.
3.8.1.8 dimer
-
 Pseudomonas sp.

Synonyms

EC Number Synonyms Comment Organism
3.5.4.43 AtzB
-
 Pseudomonas sp.
3.5.4.43 hydroxyatrazine N-ethylaminohydrolase
-
 Pseudomonas sp.
3.5.4.43 More the enzyme belongs to the amidohydrolase superfamily Pseudomonas sp.
3.8.1.8 AtzB
-
 Pseudomonas sp.
3.8.1.8 hydroxyatrazine N-ethylaminohydrolase
-
 Pseudomonas sp.

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.4.43 23
-
 assay at Pseudomonas sp.
3.8.1.8 23
-
 activity assay 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 0.8
-
 2,4-diisopropylamino-6-hydroxy-1,3,5-triazine deamination Pseudomonas sp.
3.8.1.8 1.4
-
 6-hydroxy-4-(N-isopropylamino)-2-(N-methylamino)-1,3,5-triazine deamination Pseudomonas sp.
3.8.1.8 1.6
-
 2-chloro-4-(N-ethylamino)-6-hydroxy-1,3,5-triazine dechlorination Pseudomonas sp.
3.8.1.8 1.9
-
 2-chloro-4-hydroxy-6-(N-isopropylamino)-1,3,5-triazine dechlorination Pseudomonas sp.
3.8.1.8 3
-
 hydroxyatrazine
-
 Pseudomonas sp.
3.8.1.8 3.2
-
 2-(N-ethylamino)-4-hydroxy-6-(N-isopropylamino)-1,3,5-triazine deamination Pseudomonas sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.4.43 6.5 7.5
-
 Pseudomonas sp.
3.8.1.8 7
-
 activity assay Pseudomonas sp.

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.5.4.43 6 10
-
 Pseudomonas sp.

pI Value

EC Number Organism Comment pI Value Maximum pI Value
3.5.4.43 Pseudomonas sp. isoelectric focusing, recombinant AtzB
-
 5
3.5.4.43 Pseudomonas sp. sequence calculation
-
 5.7