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.
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 |