BRENDA - Enzyme Database

Purification and characterization of a membrane-bound linoleic acid isomerase from Clostridium sporogenes

Peng, S.S.; Deng, M.; Grund, A.D.; Rosson, R.A.; Enzyme Microb. Technol. 40, 831-839 (2007)
No PubMed abstract available

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
5.2.1.5
additional information
no effects by DTT, ATP, ADP, NAD+, NADH, NADPH or CoA
Clostridium sporogenes
General Stability
EC Number
General Stability
Organism
5.2.1.5
the enzyme is very unstable, especially after being solubilized by detergents, increasing glycerol concentration to 30% and salt concentration to 0.5 M improves isomerase stability, resulting in a near full retention of activity in crude extracts for 1 week
Clostridium sporogenes
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
5.2.1.5
9-cis,12-cis-linoleic acid methyl ester
i.e. methyl linoleate, 31% inhibition at 0.02 mM
Clostridium sporogenes
5.2.1.5
9-cis,12-cis-octadecadien-1-ol
i.e. linoleyl alcohol, 19% inhibition at 0.02 mM
Clostridium sporogenes
5.2.1.5
gamma-linolenic acid
substrate inhibition at concentrations above 0.015 mM
Clostridium sporogenes
5.2.1.5
linoleic acid
substrate inhibition at concentrations above 0.02 mM, some linoleic acid derivatives also inhibited the isomerase
Clostridium sporogenes
5.2.1.5
linolelaidic acid
81% inhibition at 0.02 mM
Clostridium sporogenes
5.2.1.5
linolenic acid
substrate inhibition at concentrations above 0.04 mM
Clostridium sporogenes
5.2.1.5
additional information
no effects by DTT, ATP, ADP, NAD+, NADH, NADPH or CoA
Clostridium sporogenes
5.2.1.5
oleic acid
75% inhibition at 0.02 mM
Clostridium sporogenes
5.2.1.5
palmitoleic acid
72% inhibition at 0.02 mM
Clostridium sporogenes
5.2.1.5
stearidonic acid
substrate inhibition at concentrations above 0.120 mM
Clostridium sporogenes
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
5.2.1.5
additional information
-
additional information
-
Clostridium sporogenes
5.2.1.5
0.0034
-
6-cis,9-cis,12-cis,15-cis-octadecatetraenoic acid
pH 7.5, 22°C
Clostridium sporogenes
5.2.1.5
0.0077
-
9-cis,12-cis,15-cis-octadecatrienoic acid
pH 7.5, 22°C
Clostridium sporogenes
5.2.1.5
0.0119
-
9-cis,12-cis-linoleic acid
pH 7.5, 22°C
Clostridium sporogenes
5.2.1.5
0.0258
-
6-cis,9-cis,12-cis-octadecatrienoic acid
pH 7.5, 22°C
Clostridium sporogenes
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
5.2.1.5
membrane
bound
Clostridium sporogenes
16020
-
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
5.2.1.5
45000
-
4 * 45000, SDS-PAGE
Clostridium sporogenes
5.2.1.5
190000
-
native PAGE
Clostridium sporogenes
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
5.2.1.5
9-cis,11-trans-linoleic acid
Clostridium sporogenes
formation only under aerobic conditions
9-trans,11-trans-linoleic acid
-
-
?
5.2.1.5
9-cis,12-cis-linoleic acid
Clostridium sporogenes
higher activity under aerobic than under anaerobic conditions
9-cis,11-trans-linoleic acid
-
-
?
Organism
EC Number
Organism
UniProt
Commentary
Textmining
5.2.1.5
Clostridium sporogenes
-
strain ATCC 25762
-
Purification (Commentary)
EC Number
Purification (Commentary)
Organism
5.2.1.5
native enzyme 364fold by anion exchange chromatography, chromatofocusing, and gel filtration, method optimization for purification of the very unstable enzyme, overview
Clostridium sporogenes
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
5.2.1.5
0.4
-
purified enzyme
Clostridium sporogenes
Storage Stability
EC Number
Storage Stability
Organism
5.2.1.5
the enzyme is very unstable, especially after being solubilized by detergents, increasing glycerol concentration to 30% and salt concentration to 0.5 M improves isomerase stability, resulting in a near full retention of activity in crude extracts for 1 week
Clostridium sporogenes
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
5.2.1.5
11-cis,14-cis-eicosadienoic acid
-
679570
Clostridium sporogenes
?
-
-
-
?
5.2.1.5
6-cis,9-cis,12-cis,15-cis-octadecatetraenoic acid
i.e. stearidonic acid
679570
Clostridium sporogenes
?
-
-
-
?
5.2.1.5
6-cis,9-cis,12-cis-octadecatrienoic acid
i.e. alpha-linolenic acid
679570
Clostridium sporogenes
?
-
-
-
?
5.2.1.5
9-cis,11-trans-linoleic acid
formation only under aerobic conditions
679570
Clostridium sporogenes
9-trans,11-trans-linoleic acid
-
-
-
?
5.2.1.5
9-cis,12-cis,15-cis-octadecatrienoic acid
i.e. gamma-linolenic acid
679570
Clostridium sporogenes
?
-
-
-
?
5.2.1.5
9-cis,12-cis-linoleic acid
higher activity under aerobic than under anaerobic conditions
679570
Clostridium sporogenes
9-cis,11-trans-linoleic acid
-
-
-
?
5.2.1.5
9-cis,12-cis-linoleic acid
higher activity under aerobic than under anaerobic conditions
679570
Clostridium sporogenes
9-cis,11-trans-linoleic acid
conjugated linoleic acid isomer
-
-
?
5.2.1.5
additional information
substrate specificity, overview, the enzyme is specific for substrates containing cis double bonds at the c9 and c12 positions of C18 polyunsaturated fatty acids, a free carboxyl group is absolutely necessary for isomerization
679570
Clostridium sporogenes
?
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
5.2.1.5
tetramer
4 * 45000, SDS-PAGE
Clostridium sporogenes
Synonyms
EC Number
Synonyms
Commentary
Organism
5.2.1.5
Linoleic acid isomerase
-
Clostridium sporogenes
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
5.2.1.5
22
-
assay at room temperature
Clostridium sporogenes
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
5.2.1.5
7.5
-
-
Clostridium sporogenes
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
5.2.1.5
5.7
9
-
Clostridium sporogenes
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
5.2.1.5
additional information
no external cofactors are required for catalysis
Clostridium sporogenes
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
5.2.1.5
additional information
no effects by DTT, ATP, ADP, NAD+, NADH, NADPH or CoA
Clostridium sporogenes
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
5.2.1.5
additional information
no external cofactors are required for catalysis
Clostridium sporogenes
General Stability (protein specific)
EC Number
General Stability
Organism
5.2.1.5
the enzyme is very unstable, especially after being solubilized by detergents, increasing glycerol concentration to 30% and salt concentration to 0.5 M improves isomerase stability, resulting in a near full retention of activity in crude extracts for 1 week
Clostridium sporogenes
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
5.2.1.5
9-cis,12-cis-linoleic acid methyl ester
i.e. methyl linoleate, 31% inhibition at 0.02 mM
Clostridium sporogenes
5.2.1.5
9-cis,12-cis-octadecadien-1-ol
i.e. linoleyl alcohol, 19% inhibition at 0.02 mM
Clostridium sporogenes
5.2.1.5
gamma-linolenic acid
substrate inhibition at concentrations above 0.015 mM
Clostridium sporogenes
5.2.1.5
linoleic acid
substrate inhibition at concentrations above 0.02 mM, some linoleic acid derivatives also inhibited the isomerase
Clostridium sporogenes
5.2.1.5
linolelaidic acid
81% inhibition at 0.02 mM
Clostridium sporogenes
5.2.1.5
linolenic acid
substrate inhibition at concentrations above 0.04 mM
Clostridium sporogenes
5.2.1.5
additional information
no effects by DTT, ATP, ADP, NAD+, NADH, NADPH or CoA
Clostridium sporogenes
5.2.1.5
oleic acid
75% inhibition at 0.02 mM
Clostridium sporogenes
5.2.1.5
palmitoleic acid
72% inhibition at 0.02 mM
Clostridium sporogenes
5.2.1.5
stearidonic acid
substrate inhibition at concentrations above 0.120 mM
Clostridium sporogenes
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
5.2.1.5
additional information
-
additional information
-
Clostridium sporogenes
5.2.1.5
0.0034
-
6-cis,9-cis,12-cis,15-cis-octadecatetraenoic acid
pH 7.5, 22°C
Clostridium sporogenes
5.2.1.5
0.0077
-
9-cis,12-cis,15-cis-octadecatrienoic acid
pH 7.5, 22°C
Clostridium sporogenes
5.2.1.5
0.0119
-
9-cis,12-cis-linoleic acid
pH 7.5, 22°C
Clostridium sporogenes
5.2.1.5
0.0258
-
6-cis,9-cis,12-cis-octadecatrienoic acid
pH 7.5, 22°C
Clostridium sporogenes
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
5.2.1.5
membrane
bound
Clostridium sporogenes
16020
-
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
5.2.1.5
45000
-
4 * 45000, SDS-PAGE
Clostridium sporogenes
5.2.1.5
190000
-
native PAGE
Clostridium sporogenes
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
5.2.1.5
9-cis,11-trans-linoleic acid
Clostridium sporogenes
formation only under aerobic conditions
9-trans,11-trans-linoleic acid
-
-
?
5.2.1.5
9-cis,12-cis-linoleic acid
Clostridium sporogenes
higher activity under aerobic than under anaerobic conditions
9-cis,11-trans-linoleic acid
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
5.2.1.5
native enzyme 364fold by anion exchange chromatography, chromatofocusing, and gel filtration, method optimization for purification of the very unstable enzyme, overview
Clostridium sporogenes
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
5.2.1.5
0.4
-
purified enzyme
Clostridium sporogenes
Storage Stability (protein specific)
EC Number
Storage Stability
Organism
5.2.1.5
the enzyme is very unstable, especially after being solubilized by detergents, increasing glycerol concentration to 30% and salt concentration to 0.5 M improves isomerase stability, resulting in a near full retention of activity in crude extracts for 1 week
Clostridium sporogenes
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
5.2.1.5
11-cis,14-cis-eicosadienoic acid
-
679570
Clostridium sporogenes
?
-
-
-
?
5.2.1.5
6-cis,9-cis,12-cis,15-cis-octadecatetraenoic acid
i.e. stearidonic acid
679570
Clostridium sporogenes
?
-
-
-
?
5.2.1.5
6-cis,9-cis,12-cis-octadecatrienoic acid
i.e. alpha-linolenic acid
679570
Clostridium sporogenes
?
-
-
-
?
5.2.1.5
9-cis,11-trans-linoleic acid
formation only under aerobic conditions
679570
Clostridium sporogenes
9-trans,11-trans-linoleic acid
-
-
-
?
5.2.1.5
9-cis,12-cis,15-cis-octadecatrienoic acid
i.e. gamma-linolenic acid
679570
Clostridium sporogenes
?
-
-
-
?
5.2.1.5
9-cis,12-cis-linoleic acid
higher activity under aerobic than under anaerobic conditions
679570
Clostridium sporogenes
9-cis,11-trans-linoleic acid
-
-
-
?
5.2.1.5
9-cis,12-cis-linoleic acid
higher activity under aerobic than under anaerobic conditions
679570
Clostridium sporogenes
9-cis,11-trans-linoleic acid
conjugated linoleic acid isomer
-
-
?
5.2.1.5
additional information
substrate specificity, overview, the enzyme is specific for substrates containing cis double bonds at the c9 and c12 positions of C18 polyunsaturated fatty acids, a free carboxyl group is absolutely necessary for isomerization
679570
Clostridium sporogenes
?
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
5.2.1.5
tetramer
4 * 45000, SDS-PAGE
Clostridium sporogenes
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
5.2.1.5
22
-
assay at room temperature
Clostridium sporogenes
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
5.2.1.5
7.5
-
-
Clostridium sporogenes
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
5.2.1.5
5.7
9
-
Clostridium sporogenes