BRENDA - Enzyme Database show

Purification and characterization of 3-ketoacyl-acyl carrier protein synthase III from spinach. A condensing enzyme utilizing acetyl-coenzyme A to initiate fatty acid synthesis

Clough, R.; Matthis, A.L.; Barnum, S.R.; Jaworski, J.G.; J. Biol. Chem. 267, 20992-20998 (1992)

Data extracted from this reference:

General Stability
EC Number
General Stability
Organism
2.3.1.41
dithiothreitol stabilizes
Spinacia oleracea
2.3.1.41
glycerol stabilizes
Spinacia oleracea
2.3.1.41
Triton X-100 stabilizes
Spinacia oleracea
2.3.1.180
0.01% Triton X-100 is essential for enzyme stability during purification
Spinacia oleracea
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
2.3.1.41
arsenite
synthase III
Spinacia oleracea
2.3.1.41
N-ethylmaleimide
-
Spinacia oleracea
2.3.1.180
arsenite
45% inhibition at 5 mM, 98% inhibition at 20 mM
Spinacia oleracea
2.3.1.180
N-ethylmaleimide
complete inhibition at 5 mM
Spinacia oleracea
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.3.1.41
40500
-
2 * 40500, synthase III, SDS-PAGE
Spinacia oleracea
2.3.1.41
63000
-
synthase III, gel filtration
Spinacia oleracea
2.3.1.180
40500
-
2 * 40500, SDS-PAGE
Spinacia oleracea
2.3.1.180
63000
-
gel filtration
Spinacia oleracea
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.3.1.180
acetyl-CoA + malonyl-[acyl-carrier protein]
Spinacia oleracea
-
acetoacetyl-[acyl-carrier protein] + CoA + CO2
-
-
r
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.3.1.41
Spinacia oleracea
-
synthase III
-
2.3.1.180
Spinacia oleracea
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
2.3.1.41
synthase III, native wild-type and recombinant from E. coli E103S
Spinacia oleracea
2.3.1.180
3500fold to homogeneity from leaves in an eight-step procedure including ACP affinity chromatography
Spinacia oleracea
Source Tissue
EC Number
Source Tissue
Commentary
Organism
Textmining
2.3.1.41
leaf
-
Spinacia oleracea
-
2.3.1.180
leaf
-
Spinacia oleracea
-
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.3.1.41
additional information
-
comparison of activity, recombinant and native enzyme
Spinacia oleracea
2.3.1.41
7
-
purified native enzyme
Spinacia oleracea
2.3.1.180
7
-
purified enzyme
Spinacia oleracea
Storage Stability
EC Number
Storage Stability
Organism
2.3.1.41
-80°C, several weeks in the presence of 20% v/v glycerol, 0.1% Triton X-100 and dithiothreitol
Spinacia oleracea
2.3.1.180
-80°C, purified enzyme, 20 mM Tris buffer, 1 mM DTT, 20% glycerol, 0.01% Triton X-100, several weeks with slight loss of activity
Spinacia oleracea
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.3.1.41
acetyl-[acyl-carrier protein] + malonyl-[acyl-carrier protein]
synthase III, recombinant enzyme from E. coli is absolutely specific for
486931
Spinacia oleracea
acetoacyl-[acyl-carrier protein] + CO2 + [acyl-carrier protein]
-
486931
Spinacia oleracea
r
2.3.1.41
saturated acyl-[acyl-carrier protein] + malonyl-[acyl-carrier protein]
-
486931
Spinacia oleracea
3-oxoacyl-[acyl-carrier protein] + CO2 + [acyl-carrier protein]
-
486931
Spinacia oleracea
r
2.3.1.180
acetyl-CoA + malonyl-[acyl-carrier protein]
-
486931
Spinacia oleracea
acetoacetyl-[acyl-carrier protein] + CoA + CO2
-
-
-
r
2.3.1.180
additional information
substrate specificity, the enzyme also shows acetyltransferase activity, which is lower than the 3-ketoacyl carrier protein synthase activity
486931
Spinacia oleracea
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
Organism
2.3.1.41
dimer
2 * 40500, synthase III, SDS-PAGE
Spinacia oleracea
2.3.1.180
dimer
2 * 40500, SDS-PAGE
Spinacia oleracea
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.3.1.180
32
-
assay at
Spinacia oleracea
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.3.1.180
8
-
assay at
Spinacia oleracea
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
2.3.1.180
CoA
-
Spinacia oleracea
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
2.3.1.180
CoA
-
Spinacia oleracea
General Stability (protein specific)
EC Number
General Stability
Organism
2.3.1.41
dithiothreitol stabilizes
Spinacia oleracea
2.3.1.41
glycerol stabilizes
Spinacia oleracea
2.3.1.41
Triton X-100 stabilizes
Spinacia oleracea
2.3.1.180
0.01% Triton X-100 is essential for enzyme stability during purification
Spinacia oleracea
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
2.3.1.41
arsenite
synthase III
Spinacia oleracea
2.3.1.41
N-ethylmaleimide
-
Spinacia oleracea
2.3.1.180
arsenite
45% inhibition at 5 mM, 98% inhibition at 20 mM
Spinacia oleracea
2.3.1.180
N-ethylmaleimide
complete inhibition at 5 mM
Spinacia oleracea
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.3.1.41
40500
-
2 * 40500, synthase III, SDS-PAGE
Spinacia oleracea
2.3.1.41
63000
-
synthase III, gel filtration
Spinacia oleracea
2.3.1.180
40500
-
2 * 40500, SDS-PAGE
Spinacia oleracea
2.3.1.180
63000
-
gel filtration
Spinacia oleracea
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.3.1.180
acetyl-CoA + malonyl-[acyl-carrier protein]
Spinacia oleracea
-
acetoacetyl-[acyl-carrier protein] + CoA + CO2
-
-
r
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.3.1.41
synthase III, native wild-type and recombinant from E. coli E103S
Spinacia oleracea
2.3.1.180
3500fold to homogeneity from leaves in an eight-step procedure including ACP affinity chromatography
Spinacia oleracea
Source Tissue (protein specific)
EC Number
Source Tissue
Commentary
Organism
Textmining
2.3.1.41
leaf
-
Spinacia oleracea
-
2.3.1.180
leaf
-
Spinacia oleracea
-
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.3.1.41
additional information
-
comparison of activity, recombinant and native enzyme
Spinacia oleracea
2.3.1.41
7
-
purified native enzyme
Spinacia oleracea
2.3.1.180
7
-
purified enzyme
Spinacia oleracea
Storage Stability (protein specific)
EC Number
Storage Stability
Organism
2.3.1.41
-80°C, several weeks in the presence of 20% v/v glycerol, 0.1% Triton X-100 and dithiothreitol
Spinacia oleracea
2.3.1.180
-80°C, purified enzyme, 20 mM Tris buffer, 1 mM DTT, 20% glycerol, 0.01% Triton X-100, several weeks with slight loss of activity
Spinacia oleracea
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.3.1.41
acetyl-[acyl-carrier protein] + malonyl-[acyl-carrier protein]
synthase III, recombinant enzyme from E. coli is absolutely specific for
486931
Spinacia oleracea
acetoacyl-[acyl-carrier protein] + CO2 + [acyl-carrier protein]
-
486931
Spinacia oleracea
r
2.3.1.41
saturated acyl-[acyl-carrier protein] + malonyl-[acyl-carrier protein]
-
486931
Spinacia oleracea
3-oxoacyl-[acyl-carrier protein] + CO2 + [acyl-carrier protein]
-
486931
Spinacia oleracea
r
2.3.1.180
acetyl-CoA + malonyl-[acyl-carrier protein]
-
486931
Spinacia oleracea
acetoacetyl-[acyl-carrier protein] + CoA + CO2
-
-
-
r
2.3.1.180
additional information
substrate specificity, the enzyme also shows acetyltransferase activity, which is lower than the 3-ketoacyl carrier protein synthase activity
486931
Spinacia oleracea
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
2.3.1.41
dimer
2 * 40500, synthase III, SDS-PAGE
Spinacia oleracea
2.3.1.180
dimer
2 * 40500, SDS-PAGE
Spinacia oleracea
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.3.1.180
32
-
assay at
Spinacia oleracea
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.3.1.180
8
-
assay at
Spinacia oleracea