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

BRENDA support

Literature summary extracted from

  • Lack, N.; Yam, K.; Lowe, E.; Horsman, G.; Owen, R.; Sim, E.; Eltis, L.
    Characterization of a carbon-carbon hydrolase from Mycobacterium tuberculosis involved in cholesterol metabolism (2010), J. Biol. Chem., 285, 434-443.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.7.1.17 expressed in Escherichia coli strain GJ1158 Mycobacterium tuberculosis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.7.1.8 mutant S114A in complex with 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid, 8-(2-chlorophenyl)-2-hydroxy-5-methyl-6-oxoocta-2,4-dienoic acid, or 4,5-9,10-diseco-3-hydroxy-5,9,17-trioxoandrosta-1(10),2-diene-4-oic acid, sitting drop vapor diffusion, 0.001 ml of 8 mg/ml protein solution is mixed with 0.001 ml of precipitant solution containing 200 mM KSCN, 24% PEG 3350, and 100 mM bis-tris propane, pH 7.0, soaking of S114A crystals in 0.01 ml of precipitant solution supplemented with 15 mM of each ligand for 30-60 min, X-ray diffractin structure determination and analysis at 1.8-2.1 A resolution, molecular replacement Mycobacterium tuberculosis
3.7.1.17 mutant enzyme S114A, sitting drop vapor diffusion method, using 200 mM KSCN, 24% polyethylene glycol 3.35 K, and 100 mM bis-tris propane, pH 7.0 Mycobacterium tuberculosis

Protein Variants

EC Number Protein Variants Comment Organism
3.7.1.8 S114A active site HsaD mutant, catalytically impaired and binds 4,5-9,10-diseco-3-hydroxy-5,9,17-trioxoandrosta-1(10),2-diene-4-oic acid with an altered dissociation constant compared to the wild-type enzyme Mycobacterium tuberculosis
3.7.1.17 S114A the mutant has a 6500fold lower kcat for 4,5–9,10-diseco-3-hydroxy-5,9,17-tri-oxoandrosta-1(10),2-diene-4-oic acid compared to the wild type enzyme Mycobacterium tuberculosis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.7.1.8 0.008
-
 2-Hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid pH 7.5, 25°C Mycobacterium tuberculosis
3.7.1.8 0.31
-
 8-(2-chlorophenyl)-2-hydroxy-5-methyl-6-oxoocta-2,4-dienoic acid pH 7.5, 25°C Mycobacterium tuberculosis
3.7.1.17 0.008
-
 2-Hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid wild type enzyme, in 100 mM potassium phosphate, pH 7.5, at 25°C Mycobacterium tuberculosis
3.7.1.17 0.017
-
 4,5-9,10-diseco-3-hydroxy-5,9,17-tri-oxoandrosta-1(10),2-diene-4-oic acid wild type enzyme, in 100 mM potassium phosphate, pH 7.5, at 25°C Mycobacterium tuberculosis
3.7.1.17 0.31
-
 8-(2-chlorophenyl)-2-hydroxy-5-methyl-6-oxoocta-2,4-dienoic acid wild type enzyme, in 100 mM potassium phosphate, pH 7.5, at 25°C Mycobacterium tuberculosis

Organism

EC Number Organism UniProt Comment Textmining
3.7.1.8 Mycobacterium tuberculosis P9WNH5
-
-
3.7.1.8 Mycobacterium tuberculosis H37Rv P9WNH5
-
-
3.7.1.17 Mycobacterium tuberculosis
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.7.1.17 Source 15Q anion exchange resin column chromatography Mycobacterium tuberculosis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.7.1.8 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid + H2O i.e. HOPDA, a biphenyl meta-cleavage product Mycobacterium tuberculosis 2-hydroxypenta-2,4-dienoic acid + benzoate
-
 ?
3.7.1.8 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid + H2O i.e. HOPDA, a biphenyl meta-cleavage product Mycobacterium tuberculosis H37Rv 2-hydroxypenta-2,4-dienoic acid + benzoate
-
 ?
3.7.1.8 4,5-9,10-diseco-3-hydroxy-5,9,17-trioxoandrosta-1(10),2-diene-4-oic acid + H2O i.e. DSHA, a cholesterol meta-cleavage product Mycobacterium tuberculosis 3-[(7aR)-7a-hydroxy-1,5-dioxooctahydro-1H-inden-4-yl]propanoic acid + (2Z,4Z)-5-hydroxyhexa-2,4-dienoic acid
-
 ?
3.7.1.8 4,5-9,10-diseco-3-hydroxy-5,9,17-trioxoandrosta-1(10),2-diene-4-oic acid + H2O i.e. DSHA, a cholesterol meta-cleavage product Mycobacterium tuberculosis H37Rv 3-[(7aR)-7a-hydroxy-1,5-dioxooctahydro-1H-inden-4-yl]propanoic acid + (2Z,4Z)-5-hydroxyhexa-2,4-dienoic acid
-
 ?
3.7.1.8 8-(2-chlorophenyl)-2-hydroxy-5-methyl-6-oxoocta-2,4-dienoic acid + H2O i.e. HOPODA, a HOPDA synthetic analogue Mycobacterium tuberculosis 2-hydroxypenta-2,4-dienoic acid + benzoate
-
 ?
3.7.1.8 8-(2-chlorophenyl)-2-hydroxy-5-methyl-6-oxoocta-2,4-dienoic acid + H2O i.e. HOPODA, a HOPDA synthetic analogue Mycobacterium tuberculosis H37Rv 2-hydroxypenta-2,4-dienoic acid + benzoate
-
 ?
3.7.1.17 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid + H2O
-
 Mycobacterium tuberculosis ?
-
 ?
3.7.1.17 4,5-9,10-diseco-3-hydroxy-5,9,17-tri-oxoandrosta-1(10),2-diene-4-oic acid + H2O best substrate Mycobacterium tuberculosis 9,17-dioxo-1,2,3,4,10,19-hexanorandrostan-5-oic acid + 2-hydroxy-hexa-2,4-dienoic acid
-
 ?
3.7.1.17 8-(2-chlorophenyl)-2-hydroxy-5-methyl-6-oxoocta-2,4-dienoic acid + H2O
-
 Mycobacterium tuberculosis ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.7.1.8 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate hydrolase
-
 Mycobacterium tuberculosis
3.7.1.8 HsaD
-
 Mycobacterium tuberculosis
3.7.1.17 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate hydrolase
-
 Mycobacterium tuberculosis
3.7.1.17 HsaD
-
 Mycobacterium tuberculosis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.7.1.8 25
-
 assay at Mycobacterium tuberculosis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.7.1.8 0.066
-
 2-Hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid pH 7.5, 25°C Mycobacterium tuberculosis
3.7.1.8 0.33
-
 8-(2-chlorophenyl)-2-hydroxy-5-methyl-6-oxoocta-2,4-dienoic acid pH 7.5, 25°C Mycobacterium tuberculosis
3.7.1.17 0.000085
-
 4,5-9,10-diseco-3-hydroxy-5,9,17-tri-oxoandrosta-1(10),2-diene-4-oic acid mutant enzyme S114A, in 100 mM potassium phosphate, pH 7.5, at 25°C Mycobacterium tuberculosis
3.7.1.17 0.066
-
 2-Hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid wild type enzyme, in 100 mM potassium phosphate, pH 7.5, at 25°C Mycobacterium tuberculosis
3.7.1.17 0.33
-
 8-(2-chlorophenyl)-2-hydroxy-5-methyl-6-oxoocta-2,4-dienoic acid wild type enzyme, in 100 mM potassium phosphate, pH 7.5, at 25°C Mycobacterium tuberculosis
3.7.1.17 0.55
-
 4,5-9,10-diseco-3-hydroxy-5,9,17-tri-oxoandrosta-1(10),2-diene-4-oic acid wild type enzyme, in 100 mM potassium phosphate, pH 7.5, at 25°C Mycobacterium tuberculosis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.7.1.8 7.5
-
 assay at Mycobacterium tuberculosis

General Information

EC Number General Information Comment Organism
3.7.1.8 physiological function the enzyme is involved in the intracellular survival of pathogen Mycobacterium tuberculosis Mycobacterium tuberculosis

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.7.1.8 1.1
-
 8-(2-chlorophenyl)-2-hydroxy-5-methyl-6-oxoocta-2,4-dienoic acid pH 7.5, 25°C Mycobacterium tuberculosis
3.7.1.8 9
-
 2-Hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid pH 7.5, 25°C Mycobacterium tuberculosis
3.7.1.17 1.06
-
 8-(2-chlorophenyl)-2-hydroxy-5-methyl-6-oxoocta-2,4-dienoic acid wild type enzyme, in 100 mM potassium phosphate, pH 7.5, at 25°C Mycobacterium tuberculosis
3.7.1.17 9
-
 2-Hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid wild type enzyme, in 100 mM potassium phosphate, pH 7.5, at 25°C Mycobacterium tuberculosis
3.7.1.17 33
-
 4,5-9,10-diseco-3-hydroxy-5,9,17-tri-oxoandrosta-1(10),2-diene-4-oic acid wild type enzyme, in 100 mM potassium phosphate, pH 7.5, at 25°C Mycobacterium tuberculosis