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Literature summary for 3.7.1.17 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
Search for more literature for 3.7.1.17

Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment) Organism
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

Protein Variants Comment Organism
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]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
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
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
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

Organism UniProt Comment Textmining
Mycobacterium tuberculosis
-
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid + H2O
-
 Mycobacterium tuberculosis ?
-
 ?
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
-
 ?
8-(2-chlorophenyl)-2-hydroxy-5-methyl-6-oxoocta-2,4-dienoic acid + H2O
-
 Mycobacterium tuberculosis ?
-
 ?

Synonyms

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

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
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
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
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
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

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
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
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
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