Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • van Leeuwen, J.G.; Wijma, H.J.; Floor, R.J.; van der Laan, J.M.; Janssen, D.B.
    Directed evolution strategies for enantiocomplementary haloalkane dehalogenases: from chemical waste to enantiopure building blocks (2012), ChemBioChem, 13, 137-148.
    View publication on PubMed

Application

EC Number Application Comment Organism
3.8.1.5 environmental protection DhaA is capable of degrading 1,2,3-trichloropropane, TCP, an industrial waste product that is toxic, extremely recalcitrant to biodegradation, and expensive to dispose of by physical or chemical methods Rhodococcus rhodochrous
3.8.1.5 synthesis DhaA produces reaction products (R)- and (S)-2,3-dichloropropan-1-ol, which can be converted to (S)- and (R)-epihydrins, valuable fine chemicals that find application in synthetic routes to several pharmaceutical and healthcare products Rhodococcus rhodochrous

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.8.1.5 recombinant expression of His-tagged wild-type and mutant enzymes Rhodococcus rhodochrous

Protein Variants

EC Number Protein Variants Comment Organism
3.8.1.5 F168W/A172L/Y176G site-directed mutagenesis, the mutant shows increased enantioselectivity with substrate TCP compared to the wild-type enzyme, 1,2,3-trichloropropane is docked in the active site in a configuration that leads to (R)-2,3-dichloropropan-1-ol formation Rhodococcus rhodochrous
3.8.1.5 I135F/C176Y/V245F/L246I/Y273F structural modeling of the mutant compared to the wild-type DhaA31 using the wild-type crystal structure Rhodococcus rhodochrous
3.8.1.5 additional information library screening for mutants with altered enantioselectivity with substrate 1,2,3-trichloropropane compared to DhaA31 wild-type, overview Rhodococcus rhodochrous

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.8.1.5 additional information
-
additional information steady-state kinetics with substrate1,2,3-trichloropropane, wild-type and mutant enzymes, overview Rhodococcus rhodochrous

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.8.1.5 1,2,3-trichloropropane + H2O Rhodococcus rhodochrous
-
(RS)-2,3-dichloropropan-1-ol + chloride
-
?

Organism

EC Number Organism UniProt Comment Textmining
3.8.1.5 Rhodococcus rhodochrous
-
gene dhaA
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.8.1.5 recombinant His-tagged wild-type and mutant enzymes by nickel affinity chromatography Rhodococcus rhodochrous

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.8.1.5 1,2,3-trichloropropane + H2O
-
Rhodococcus rhodochrous (RS)-2,3-dichloropropan-1-ol + chloride
-
?
3.8.1.5 1,2,3-trichloropropane + H2O structural model of DhaA31 with TCP docked in the active site, comparison with DhaA31 mutant enzyme docking models, overview. The pentad is made up of an Asp-His-Asp catalytic triad and a Trp-Trp or Trp-Asn diad for halide binding. An aspartate residue acts as the nucleophile to displace a halide ion from the substrate. By hydrolysis of the resulting covalent alkyl-enzyme intermediate, alcohol is released Rhodococcus rhodochrous (RS)-2,3-dichloropropan-1-ol + chloride
-
?

Subunits

EC Number Subunits Comment Organism
3.8.1.5 More the enzyme is composed of a main domain and a cap domain, with a catalytic pentad located between these domains Rhodococcus rhodochrous

Synonyms

EC Number Synonyms Comment Organism
3.8.1.5 DhaA
-
Rhodococcus rhodochrous
3.8.1.5 DhaA31
-
Rhodococcus rhodochrous

General Information

EC Number General Information Comment Organism
3.8.1.5 evolution the haloalkane dehalogenases form a subclass of enzymes in the alpha/beta-hydrolase fold superfamily Rhodococcus rhodochrous