BRENDA - Enzyme Database show
show all sequences of 3.7.1.17

Characterization of a carbon-carbon hydrolase from Mycobacterium tuberculosis involved in cholesterol metabolism

Lack, N.; Yam, K.; Lowe, E.; Horsman, G.; Owen, R.; Sim, E.; Eltis, L.; J. Biol. Chem. 285, 434-443 (2010)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expressed in Escherichia coli strain GJ1158
Mycobacterium tuberculosis
Crystallization (Commentary)
Crystallization
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
Engineering
Amino acid exchange
Commentary
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
Commentary
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
Primary Accession No. (UniProt)
Commentary
Textmining
Mycobacterium tuberculosis
-
-
-
Purification (Commentary)
Commentary
Organism
Source 15Q anion exchange resin column chromatography
Mycobacterium tuberculosis
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid + H2O
-
717815
Mycobacterium tuberculosis
?
-
-
-
?
4,5-9,10-diseco-3-hydroxy-5,9,17-tri-oxoandrosta-1(10),2-diene-4-oic acid + H2O
best substrate
717815
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
-
717815
Mycobacterium tuberculosis
?
-
-
-
?
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
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
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed in Escherichia coli strain GJ1158
Mycobacterium tuberculosis
Crystallization (Commentary) (protein specific)
Crystallization
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
Engineering (protein specific)
Amino acid exchange
Commentary
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] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
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
Purification (Commentary) (protein specific)
Commentary
Organism
Source 15Q anion exchange resin column chromatography
Mycobacterium tuberculosis
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid + H2O
-
717815
Mycobacterium tuberculosis
?
-
-
-
?
4,5-9,10-diseco-3-hydroxy-5,9,17-tri-oxoandrosta-1(10),2-diene-4-oic acid + H2O
best substrate
717815
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
-
717815
Mycobacterium tuberculosis
?
-
-
-
?
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
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
Commentary
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
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
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
Other publictions for EC 3.7.1.17
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
733885
Ryan
Mechanism-based inhibition of ...
Mycobacterium tuberculosis
FEMS Microbiol. Lett.
350
42-47
2014
-
-
1
-
-
-
14
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
-
1
-
-
-
1
-
-
-
-
-
1
-
-
1
-
-
-
-
1
14
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
717815
Lack
Characterization of a carbon-c ...
Mycobacterium tuberculosis
J. Biol. Chem.
285
434-443
2010
-
-
1
1
1
-
-
3
-
-
-
-
-
1
-
-
1
-
-
-
-
-
3
-
-
-
-
4
-
-
-
-
-
-
-
-
-
1
-
1
1
-
-
-
-
3
-
-
-
-
-
-
-
1
-
-
-
-
3
-
-
-
-
4
-
-
-
-
-
-
-
-
3
3
717209
Lack
Temperature stability of prote ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
Biochem. J.
418
369-378
2009
-
-
1
-
-
-
-
1
1
-
2
-
-
152
-
-
1
-
-
-
-
-
2
1
-
-
1
1
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
1
1
-
2
-
-
-
-
1
-
-
-
-
2
1
-
-
1
1
-
-
-
-
-
-
-
-
1
1
717030
Lack
Structure of HsaD, a steroid-d ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
Acta Crystallogr. Sect. F
64
2-7
2007
-
-
1
1
-
-
-
-
-
-
-
2
-
152
-
-
1
-
-
-
-
-
2
1
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
-
2
-
-
-
1
-
-
-
-
2
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
718315
Van Der Geize
A gene cluster encoding choles ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
Proc. Natl. Acad. Sci. USA
104
1947-1952
2007
-
-
1
-
-
-
-
1
-
-
-
-
-
152
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
654270
Horinouchi
A new bacterial steroid degrad ...
Comamonas testosteroni, Comamonas testosteroni TA441
Appl. Environ. Microbiol.
69
4421-4430
2003
-
-
-
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-