Literature summary extracted from

  • Laempe, D.; Jahn, M.; Fuchs, G.
    6-Hydroxycyclohex-1-ene-1-carbonyl-CoA dehydrogenase and 6-oxocyclohex-1-ene-1-carbonyl-CoA hydrolase, enzymes of the benzoyl-CoA pathway of anaerobic aromatic metabolism in the denitrifying bacterium Thauera aromatica (1999), Eur. J. Biochem., 263, 420-429.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.368 overexpressed in Escherichia coli Thauera aromatica
3.7.1.21 overexpressed in Escherichia coli Thauera aromatica

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.368 additional information the enzymew is insensitive to 10 mM EDTA, 1 mM 2,2'-bipyridyl, or 1 mM pyrazole Thauera aromatica

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.368 0.06
-
6-hydroxycyclohex-1-ene-1-carbonyl-CoA pH 7.4, 37°C Thauera aromatica

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.368 additional information activity is not increased after adding 2-20 mM of ZnCl2 Thauera aromatica

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.368 78000
-
gel filtration Thauera aromatica
3.7.1.21 170000
-
-
Thauera aromatica
1.1.1.368 38000
-
x * 38000, SDS-PAGE Thauera aromatica
3.7.1.21 40000
-
4 * 40000, SDS-PAGE Thauera aromatica

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.368 6-hydroxycyclohex-1-ene-1-carbonyl-CoA + NAD+ Thauera aromatica the enzyme participates in the central benzoyl-CoA degradation pathway of some anaerobic bacteria 6-oxocyclohex-1-ene-1-carbonyl-CoA + NADH + H+
-
ir
3.7.1.21 6-oxocyclohex-1-ene-1-carbonyl-CoA + 2 H2O Thauera aromatica the enzyme participates in the anaerobic benzoyl-CoA degradation pathway 3-hydroxypimeloyl-CoA 3-hydroxypimeloyl-CoA i.e. 3-hydroxy-6-carboxyhexanoyl-CoA ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.368 Thauera aromatica O87871
-
-
3.7.1.21 Thauera aromatica O87872
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.368 recombinant protein Thauera aromatica
3.7.1.21
-
Thauera aromatica

Source Tissue

EC Number Source Tissue Comment Organism Textmining
1.1.1.368 culture condition when cells are grown aerobically with benzoate the activity is 20-25fold lower than in cells grown anaerobically with benzoate Thauera aromatica
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.368 11.8
-
pH 7.4, 37°C, substrate: 6-hydroxycyclohex-1-ene-1-carbonyl-CoA Thauera aromatica
3.7.1.21 1.3
-
pH 7.4, 37°C, substrate: 6-oxocyclohex-1-ene-1-carbonyl-CoA Thauera aromatica

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.368 6-hydroxycyclohex-1-ene-1-carbonyl-CoA + NAD+ the enzyme participates in the central benzoyl-CoA degradation pathway of some anaerobic bacteria Thauera aromatica 6-oxocyclohex-1-ene-1-carbonyl-CoA + NADH + H+
-
ir
1.1.1.368 6-hydroxycyclohex-1-ene-1-carbonyl-CoA + NAD+ the enzyme is specific for NAD+, no activity with NADP+. The enzyme is specific for 6-hydroxycyclohex-1-ene-1-carbonyl-CoA, no activity with 2-hydroxycyclohexane-1-carbonyl-CoA Thauera aromatica 6-oxocyclohex-1-ene-1-carbonyl-CoA + NADH + H+
-
ir
3.7.1.21 6-oxocyclohex-1-ene-1-carbonyl-CoA + 2 H2O the enzyme participates in the anaerobic benzoyl-CoA degradation pathway Thauera aromatica 3-hydroxypimeloyl-CoA 3-hydroxypimeloyl-CoA i.e. 3-hydroxy-6-carboxyhexanoyl-CoA ?
3.7.1.21 6-oxocyclohex-1-ene-1-carbonyl-CoA + 2 H2O the enzyme catalyses the addition of one molecule of water to the double bound of 6-oxocyclohex-1-ene-1-carbonyl-CoA followed by the hydrolytic C-C cleavage of the alicyclic ring Thauera aromatica 3-hydroxypimeloyl-CoA 3-hydroxypimeloyl-CoA i.e. 3-hydroxy-6-carboxyhexanoyl-CoA ?

Subunits

EC Number Subunits Comment Organism
1.1.1.368 ? x * 38000, SDS-PAGE Thauera aromatica
3.7.1.21 tetramer 4 * 40000, SDS-PAGE Thauera aromatica

Synonyms

EC Number Synonyms Comment Organism
1.1.1.368 HAD
-
Thauera aromatica
3.7.1.21 6-oxocyclohex-1-ene-1-carbonyl-CoA hydrolase
-
Thauera aromatica

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.368 37
-
assay at Thauera aromatica
3.7.1.21 37
-
assay at Thauera aromatica

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.368 7
-
6-hydroxycyclohex-1-ene-1-carbonyl-CoA pH 7.4, 37°C, calculated as kcat number per subunit Thauera aromatica
3.7.1.21 additional information
-
6-oxocyclohex-1-ene-1-carbonyl-CoA 0.9/s per subunit Thauera aromatica

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.368 7.4
-
assay at Thauera aromatica
1.1.1.368 7.2 7.4
-
Thauera aromatica
3.7.1.21 7.4
-
assay at Thauera aromatica

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.368 NAD+ the enzyme is specific for NAD+, no activity with NADP+ Thauera aromatica

Expression

EC Number Organism Comment Expression
1.1.1.368 Thauera aromatica gene expression is positively controlled by benzoate, an intermediate of the central benzoyl-CoA pathway up
1.1.1.368 Thauera aromatica expression of the enzyme activity is under negative control by oxygen down

General Information

EC Number General Information Comment Organism
3.7.1.21 physiological function the enzyme participates in the anaerobic benzoyl-CoA degradation pathway Thauera aromatica