Literature summary extracted from

  • Tiedt, O.; Mergelsberg, M.; Eisenreich, W.; Boll, M.
    Promiscuous defluorinating enoyl-CoA hydratases/hydrolases allow for complete anaerobic degradation of 2-fluorobenzoate (2017), Front. Microbiol., 8, 2579 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.1.100 gene dch, recombinant expression of C-terminally His6-tagged enzyme in Escherichia coli strain MC4100 Thauera aromatica

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.2.1.100 2-fluorocyclohexa-1,5-diene-1-carbonyl-CoA + H2O Thauera aromatica
-
2-fluoro-6-hydroxycyclohexa-1-ene-1-carbonyl-CoA
-
r
4.2.1.100 6-fluorocyclohexa-1,5-diene-1-carbonyl-CoA + H2O Thauera aromatica
-
6-fluoro-6-hydroxycyclohexa-1-ene-1-carbonyl-CoA
-
r
4.2.1.100 6-hydroxycyclohexa-1-ene-1-carbonyl-CoA Thauera aromatica
-
cyclohexa-1,5-diene-1-carbonyl-CoA + H2O
-
r
4.2.1.100 cyclohexa-1,5-diene-1-carbonyl-CoA + H2O Thauera aromatica
-
6-hydroxycyclohexa-1-ene-1-carbonyl-CoA
-
r
4.2.1.100 2-fluorocyclohexa-1,5-diene-1-carbonyl-CoA + H2O Thauera aromatica K172
-
2-fluoro-6-hydroxycyclohexa-1-ene-1-carbonyl-CoA
-
r
4.2.1.100 6-fluorocyclohexa-1,5-diene-1-carbonyl-CoA + H2O Thauera aromatica K172
-
6-fluoro-6-hydroxycyclohexa-1-ene-1-carbonyl-CoA
-
r
4.2.1.100 6-hydroxycyclohexa-1-ene-1-carbonyl-CoA Thauera aromatica K172
-
cyclohexa-1,5-diene-1-carbonyl-CoA + H2O
-
r

Organism

EC Number Organism UniProt Comment Textmining
4.2.1.100 Thauera aromatica o87873
-
-
4.2.1.100 Thauera aromatica K172 o87873
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.1.100 recombinant His6-tagged enzyme from Escherichia coli strain MC4100 by nickel affinity chromatography and desalting gel filtration Thauera aromatica

Source Tissue

EC Number Source Tissue Comment Organism Textmining
4.2.1.100 additional information 2-fluoro-benzoate- and benzoate-grown cells Thauera aromatica
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.2.1.100 110
-
purified recombinant enzyme with substrate 6-hydroxycyclohex-1-ene-1-carbonyl-CoA, pH and temperature not specified in the publication Thauera aromatica
4.2.1.100 107
-
purified recombinant enzyme with substrate cyclohexa-1,5-diene-1-carbonyl-CoA, pH and temperature not specified in the publication Thauera aromatica

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.100 additional information the 1,5-dienoyl-CoA hydratase hydrated the fluorocyclohexa-1,5-dienoyl-CoA isomers to a mixture of the stable 2-fluoro-6-hydroxycyclohexa-1-ene-1-carbonyl-CoA and the unstable alpha-fluorohydrin 6-fluoro-6-hydroxycyclohexa-1-ene-1-carbonyl-CoA. The latter spontaneously decomposed to HF and 6-oxo-cyclohexa-1-ene-1-carbonyl-CoA Thauera aromatica ?
-
?
4.2.1.100 additional information the 1,5-dienoyl-CoA hydratase hydrated the fluorocyclohexa-1,5-dienoyl-CoA isomers to a mixture of the stable 2-fluoro-6-hydroxycyclohexa-1-ene-1-carbonyl-CoA and the unstable alpha-fluorohydrin 6-fluoro-6-hydroxycyclohexa-1-ene-1-carbonyl-CoA. The latter spontaneously decomposed to HF and 6-oxo-cyclohexa-1-ene-1-carbonyl-CoA Thauera aromatica K172 ?
-
?
4.2.1.100 cyclohexa-1,5-diene-1-carbonyl-CoA + H2O
-
Thauera aromatica 6-hydroxycyclohex-1-ene-1-carbonyl-CoA
-
r
4.2.1.100 cyclohexa-1,5-diene-1-carbonyl-CoA + H2O
-
Thauera aromatica K172 6-hydroxycyclohex-1-ene-1-carbonyl-CoA
-
r
4.2.1.100 2-fluorocyclohexa-1,5-diene-1-carbonyl-CoA + H2O
-
Thauera aromatica 2-fluoro-6-hydroxycyclohexa-1-ene-1-carbonyl-CoA
-
r
4.2.1.100 2-fluorocyclohexa-1,5-diene-1-carbonyl-CoA + H2O
-
Thauera aromatica K172 2-fluoro-6-hydroxycyclohexa-1-ene-1-carbonyl-CoA
-
r
4.2.1.100 6-fluorocyclohexa-1,5-diene-1-carbonyl-CoA + H2O
-
Thauera aromatica 6-fluoro-6-hydroxycyclohexa-1-ene-1-carbonyl-CoA
-
r
4.2.1.100 6-fluorocyclohexa-1,5-diene-1-carbonyl-CoA + H2O
-
Thauera aromatica K172 6-fluoro-6-hydroxycyclohexa-1-ene-1-carbonyl-CoA
-
r
4.2.1.100 6-hydroxycyclohexa-1-ene-1-carbonyl-CoA
-
Thauera aromatica cyclohexa-1,5-diene-1-carbonyl-CoA + H2O
-
r
4.2.1.100 6-hydroxycyclohexa-1-ene-1-carbonyl-CoA
-
Thauera aromatica K172 cyclohexa-1,5-diene-1-carbonyl-CoA + H2O
-
r
4.2.1.100 cyclohexa-1,5-diene-1-carbonyl-CoA + H2O
-
Thauera aromatica 6-hydroxycyclohexa-1-ene-1-carbonyl-CoA
-
r

Synonyms

EC Number Synonyms Comment Organism
4.2.1.100 DCH
-
Thauera aromatica
4.2.1.100 1,5-dienoyl-CoA hydratase
-
Thauera aromatica

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.2.1.100 30
-
in vivo assay at Thauera aromatica

General Information

EC Number General Information Comment Organism
4.2.1.100 physiological function the enzyme is involved in the biodegradation of the environmentally hazardous fluoroaromatics. The 1,5-dienoyl-CoA hydratase (DCH) hydrates the fluorocyclohexa-1,5-diene-1-carbonyl-CoA isomers to a mixture of the stable 2-fluoro-6-hydroxycyclohexa-1-ene-1-carbonyl-CoA and the unstable alpha-fluorohydrin 6-fluoro-6-hydroxycyclohexa-1-ene-1-carbonyl-CoA. The latter spontaneously decomposes to HF and 6-oxo-cyclohexa-1-ene-1-carbonyl-CoA, a common intermediate of the benzoyl-coenzyme A (BzCoA) degradation pathway. 6-Oxo-cyclohexa-1-ene-1-carbonyl-CoA hydrolase/hydratase catalyzes the defluorination of 2-fluoro-6-hydroxy-1-enoyl-CoA to 2-oxo-6-hydroxy-1-enoyl-CoA and HF again via water addition to an fluoro-enoyl-CoA functionality Thauera aromatica
4.2.1.100 metabolism the enzyme is involved in the biodegradation of the environmentally hazardous fluoroaromatics. The 1,5-dienoyl-CoA hydratase (DCH) hydrates the fluoro-1,5-dienoyl-CoA isomers to a mixture of the stable 2-fluoro-6-hydroxy-1-enoyl-CoA and the unstable alpha-fluorohydrin 6-fluoro-6-hydroxy-1-enoyl-CoA. The latter spontaneously decomposes to HF and 6-oxo-cyclohex-1-enoyl-CoA, a common intermediate of the benzoyl-coenzyme A (BzCoA) degradation pathway. 6-Oxo-cyclohex-1-enoyl-CoA hydrolase/hydratase catalyzes the defluorination of 2-fluoro-6-hydroxycyclohex-1-enoyl-CoA to 2-oxo-6-hydroxycyclohex-1-enoyl-CoA and HF again via water addition to an fluorocyclohex-1-enoyl-CoA functionality. Formation of both 3-fluorocyclohexa-1,5-diene-1-carbonyl-CoA and 5-fluorocyclohexa-1,5-diene-1-carbonyl-CoA suggesting a low regioselectivity of benzoyl-coenzyme A (BzCoA) reductase (BCR) for fluorinated BzCoA analogues. Enzyme DCH and bifunctional 6-oxo-cyclohex-1-enoyl-CoA hydrolase (OAH) both initiate defluorination of the 2-fluorocyclohexa-1,5-diene-1-carbonyl-CoA intermediate by 1,4-hydration. Enzyme OAH is competent to hydrate both fluorinated cyclohexa-1,5-diene-carbonyl-CoA isomers and exhibits a similar reactivity as enzyme DCH, and OAH alone is capable of defluorinating both fluorinated cyclohexa-1,5-diene-carbonyl-CoA isomers formed by BCR. Detailed pathway overview Thauera aromatica