Activating Compound | Comment | Organism | Structure |
---|---|---|---|
NaCl | optimal oionic strength 0.62 mM | Burkholderia mallei |
Cloned (Comment) | Organism |
---|---|
- |
Burkholderia mallei |
gene fabV, expression of His-tagged wild-type and mutant enzymes in Escherichia coli BL21(DE3), theN-terminal His-tag in bmFabv does not have a dramatic effect on catalytic activity | Burkholderia mallei |
Protein Variants | Comment | Organism |
---|---|---|
K244A | site-directed mutagenesis | Burkholderia mallei |
K244A | mutation does not induce major structural alterations. 110fold decrease in kcat value | Burkholderia mallei |
K244A/K245A | loss of activity | Burkholderia mallei |
K244A/K245A | site-directed mutagenesis | Burkholderia mallei |
K244R | site-directed mutagenesis | Burkholderia mallei |
K244R | mutation does not induce major structural alterations. 950fold decrease in kcat value | Burkholderia mallei |
K245M | site-directed mutagenesis | Burkholderia mallei |
K245M | mutation does not induce major structural alterations. 3fold increase in Km value for substrate dodecanoyl-CoA, 70fold decrease in kcat for substrate reduction | Burkholderia mallei |
Y235A | site-directed mutagenesis | Burkholderia mallei |
Y235A | mutation does not induce major structural alterations. 280fold decrease in kcat/Km ratio | Burkholderia mallei |
Y235S | site-directed mutagenesis | Burkholderia mallei |
Y235S | mutation does not induce major structural alterations. 280fold decrease in kcat/Km ratio | Burkholderia mallei |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
triclosan | a rapid, reversible inhibitor of bmFabV, and a competitive inhibitor with respect to NADH and an uncompetitive inhibitor with respect to the substrate 2-dodecenoyl-CoA. Triclosan binds to the enzyme-NAD+ product complex which is in rapid and reversible equilibrium with other intermediates on the reaction pathway; competitive inhibitor with respect to NADH and an uncompetitive inhibitor with respect to the substrate 2-dodecenoyl-CoA | Burkholderia mallei |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Km for NADH of 0.023 mM, a Km for 2-dodecenoyl-CoA of 0.0025 mM, and a kcat of 20.7 s-1. Steady-state kinetics, overview | Burkholderia mallei | |
0.0022 | - |
(2E)-2-dodecenoyl-CoA | mutant Y235S, pH 7.9, 25°C | Burkholderia mallei | |
0.0027 | - |
(2E)-2-dodecenoyl-CoA | mutant Y235A, pH 7.9, 25°C | Burkholderia mallei | |
0.0044 | - |
(2E)-2-dodecenoyl-CoA | wild-type, pH 7.9, 25°C | Burkholderia mallei | |
0.006 | - |
NADH | mutant Y235S, pH 7.9, 25°C | Burkholderia mallei | |
0.0072 | - |
(2E)-2-dodecenoyl-CoA | mutant K244A, pH 7.9, 25°C | Burkholderia mallei | |
0.0075 | - |
(2E)-2-dodecenoyl-CoA | mutant K244R, pH 7.9, 25°C | Burkholderia mallei | |
0.023 | - |
NADH | mutant Y235A, pH 7.9, 25°C | Burkholderia mallei | |
0.023 | - |
NADH | wild-type, pH 7.9, 25°C | Burkholderia mallei | |
0.025 | - |
crotonyl-[acyl-carrier protein] | pH 7.9, 25°C | Burkholderia mallei | |
0.028 | - |
NADH | mutant K245M, pH 7.9, 25°C | Burkholderia mallei | |
0.028 | - |
(2E)-2-dodecenoyl-CoA | mutant K245M, pH 7.9, 25°C | Burkholderia mallei | |
0.066 | - |
NADH | mutant K244A, pH 7.9, 25°C | Burkholderia mallei | |
0.069 | - |
NADH | mutant K244R, pH 7.9, 25°C | Burkholderia mallei |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Burkholderia mallei | Q62L02 | - |
- |
Burkholderia mallei | Q62L02 | gene fabV | - |
Burkholderia mallei ATCC 23344 | Q62L02 | gene fabV | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli BL21(DE3) by nickel affinity chromatography | Burkholderia mallei |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
an acyl-[acyl-carrier protein] + NAD+ = a trans-2,3-dehydroacyl-[acyl-carrier protein] + NADH + H+ | detailed kinetic analysis of the sequential bi-bi mechanism with NADH binding first and NAD+ dissociating last. The catalytic tyrosine (Y235) and lysine (K244) residues are organized in the consensus Tyr-(Xaa)8-Lys motif. Both Y235 and K244 are involved in acid-base chemistry during substrate reduction | Burkholderia mallei | |
an acyl-[acyl-carrier protein] + NAD+ = a trans-2,3-dehydroacyl-[acyl-carrier protein] + NADH + H+ | sequential Bi Bi mechanism with NADH binding first and NAD+ dissociating last | Burkholderia mallei |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(2E)-2-dodecenoyl-CoA + NADH + H+ | - |
Burkholderia mallei | dodecanoyl-CoA + NAD+ | - |
? | |
(2E)-2-dodecenoyl-CoA + NADH + H+ | - |
Burkholderia mallei ATCC 23344 | dodecanoyl-CoA + NAD+ | - |
? | |
crotonyl-[acyl-carrier protein] + NAD+ | - |
Burkholderia mallei | ? + NADH + H+ | - |
? | |
crotonyl-[acyl-carrier protein] + NAD+ | - |
Burkholderia mallei ATCC 23344 | ? + NADH + H+ | - |
? | |
crotonyl-[acyl-carrier protein] + NADH | - |
Burkholderia mallei | butyryl-[acyl-carrier protein] + NAD+ | - |
? | |
crotonyl-[acyl-carrier protein] + NADH | - |
Burkholderia mallei ATCC 23344 | butyryl-[acyl-carrier protein] + NAD+ | - |
? | |
additional information | activity measurement using NAD+ cofactor and substrate 2-dodecenoyl-CoA | Burkholderia mallei | ? | - |
? | |
additional information | activity measurement using NAD+ cofactor and substrate 2-dodecenoyl-CoA | Burkholderia mallei ATCC 23344 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
BMA0885 | - |
Burkholderia mallei |
enoyl-ACP reductase | - |
Burkholderia mallei |
FabV | - |
Burkholderia mallei |
More | the enzyme is a member of the short chain dehydrogenase/reductase superfamily | Burkholderia mallei |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Burkholderia mallei |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.022 | - |
(2E)-2-dodecenoyl-CoA | mutant K244R, pH 7.9, 25°C | Burkholderia mallei | |
0.08 | - |
(2E)-2-dodecenoyl-CoA | mutant Y235A, pH 7.9, 25°C | Burkholderia mallei | |
0.1 | - |
(2E)-2-dodecenoyl-CoA | mutant Y235S, pH 7.9, 25°C | Burkholderia mallei | |
0.18 | - |
(2E)-2-dodecenoyl-CoA | mutant K244A, pH 7.9, 25°C | Burkholderia mallei | |
0.3 | - |
(2E)-2-dodecenoyl-CoA | mutant K245M, pH 7.9, 25°C | Burkholderia mallei | |
28.8 | - |
(2E)-2-dodecenoyl-CoA | wild-type, pH 7.9, 25°C | Burkholderia mallei |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.9 | - |
- |
Burkholderia mallei |
7.9 | - |
assay at | Burkholderia mallei |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6.6 | 8.5 | - |
Burkholderia mallei |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | - |
Burkholderia mallei |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | inhibition kinetics, overview | Burkholderia mallei | |
0.0004 | - |
triclosan | versus 2-dodecenoyl-CoA, pH 7.8, 25°C | Burkholderia mallei | |
0.0004 | - |
triclosan | substrate 2-dodecenoyl-CoA, pH 7.9, 25°C | Burkholderia mallei |
General Information | Comment | Organism |
---|---|---|
metabolism | enoyl-ACP reductases catalyze the final step in the elongation cycle of the bacterial fatty acid biosynthesis, FAS-II, pathway, but FabV is distinct and belongs to another class of enoyl-acyl carrier protein reductases, overview | Burkholderia mallei |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.011 | - |
(2E)-2-dodecenoyl-CoA | mutant K245M, pH 7.9, 25°C | Burkholderia mallei | |
0.025 | - |
(2E)-2-dodecenoyl-CoA | mutant K244A, pH 7.9, 25°C | Burkholderia mallei | |
0.028 | - |
(2E)-2-dodecenoyl-CoA | mutant K244R, pH 7.9, 25°C | Burkholderia mallei | |
0.03 | - |
(2E)-2-dodecenoyl-CoA | mutant Y235A, pH 7.9, 25°C | Burkholderia mallei | |
0.047 | - |
(2E)-2-dodecenoyl-CoA | mutant Y235S, pH 7.9, 25°C | Burkholderia mallei | |
6.5 | - |
(2E)-2-dodecenoyl-CoA | wild-type, pH 7.9, 25°C | Burkholderia mallei |