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Literature summary extracted from

  • Lu, H.; Tonge, P.J.
    Mechanism and inhibition of the FabV enoyl-ACP reductase from Burkholderia mallei (2010), Biochemistry, 49, 1281-1289.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.3.1.9 NaCl optimal oionic strength 0.62 mM Burkholderia mallei

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.3.1.9
-
 Burkholderia mallei
1.3.1.9 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

EC Number Protein Variants Comment Organism
1.3.1.9 K244A site-directed mutagenesis Burkholderia mallei
1.3.1.9 K244A mutation does not induce major structural alterations. 110fold decrease in kcat value Burkholderia mallei
1.3.1.9 K244A/K245A loss of activity Burkholderia mallei
1.3.1.9 K244A/K245A site-directed mutagenesis Burkholderia mallei
1.3.1.9 K244R site-directed mutagenesis Burkholderia mallei
1.3.1.9 K244R mutation does not induce major structural alterations. 950fold decrease in kcat value Burkholderia mallei
1.3.1.9 K245M site-directed mutagenesis Burkholderia mallei
1.3.1.9 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
1.3.1.9 Y235A site-directed mutagenesis Burkholderia mallei
1.3.1.9 Y235A mutation does not induce major structural alterations. 280fold decrease in kcat/Km ratio Burkholderia mallei
1.3.1.9 Y235S site-directed mutagenesis Burkholderia mallei
1.3.1.9 Y235S mutation does not induce major structural alterations. 280fold decrease in kcat/Km ratio Burkholderia mallei

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.3.1.9 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.3.1.9 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
1.3.1.9 0.0022
-
 (2E)-2-dodecenoyl-CoA mutant Y235S, pH 7.9, 25°C Burkholderia mallei
1.3.1.9 0.0027
-
 (2E)-2-dodecenoyl-CoA mutant Y235A, pH 7.9, 25°C Burkholderia mallei
1.3.1.9 0.0044
-
 (2E)-2-dodecenoyl-CoA wild-type, pH 7.9, 25°C Burkholderia mallei
1.3.1.9 0.006
-
 NADH mutant Y235S, pH 7.9, 25°C Burkholderia mallei
1.3.1.9 0.0072
-
 (2E)-2-dodecenoyl-CoA mutant K244A, pH 7.9, 25°C Burkholderia mallei
1.3.1.9 0.0075
-
 (2E)-2-dodecenoyl-CoA mutant K244R, pH 7.9, 25°C Burkholderia mallei
1.3.1.9 0.023
-
 NADH mutant Y235A, pH 7.9, 25°C Burkholderia mallei
1.3.1.9 0.023
-
 NADH wild-type, pH 7.9, 25°C Burkholderia mallei
1.3.1.9 0.025
-
 crotonyl-[acyl-carrier protein] pH 7.9, 25°C Burkholderia mallei
1.3.1.9 0.028
-
 NADH mutant K245M, pH 7.9, 25°C Burkholderia mallei
1.3.1.9 0.028
-
 (2E)-2-dodecenoyl-CoA mutant K245M, pH 7.9, 25°C Burkholderia mallei
1.3.1.9 0.066
-
 NADH mutant K244A, pH 7.9, 25°C Burkholderia mallei
1.3.1.9 0.069
-
 NADH mutant K244R, pH 7.9, 25°C Burkholderia mallei

Organism

EC Number Organism UniProt Comment Textmining
1.3.1.9 Burkholderia mallei Q62L02
-
-
1.3.1.9 Burkholderia mallei Q62L02 gene fabV
-
1.3.1.9 Burkholderia mallei ATCC 23344 Q62L02 gene fabV
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.3.1.9 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli BL21(DE3) by nickel affinity chromatography Burkholderia mallei

Reaction

EC Number Reaction Comment Organism Reaction ID
1.3.1.9 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
a
1.3.1.9 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
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.3.1.9 (2E)-2-dodecenoyl-CoA + NADH + H+
-
 Burkholderia mallei dodecanoyl-CoA + NAD+
-
 ?
1.3.1.9 (2E)-2-dodecenoyl-CoA + NADH + H+
-
 Burkholderia mallei ATCC 23344 dodecanoyl-CoA + NAD+
-
 ?
1.3.1.9 crotonyl-[acyl-carrier protein] + NAD+
-
 Burkholderia mallei ? + NADH + H+
-
 ?
1.3.1.9 crotonyl-[acyl-carrier protein] + NAD+
-
 Burkholderia mallei ATCC 23344 ? + NADH + H+
-
 ?
1.3.1.9 crotonyl-[acyl-carrier protein] + NADH
-
 Burkholderia mallei butyryl-[acyl-carrier protein] + NAD+
-
 ?
1.3.1.9 crotonyl-[acyl-carrier protein] + NADH
-
 Burkholderia mallei ATCC 23344 butyryl-[acyl-carrier protein] + NAD+
-
 ?
1.3.1.9 additional information activity measurement using NAD+ cofactor and substrate 2-dodecenoyl-CoA Burkholderia mallei ?
-
 ?
1.3.1.9 additional information activity measurement using NAD+ cofactor and substrate 2-dodecenoyl-CoA Burkholderia mallei ATCC 23344 ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.3.1.9 BMA0885
-
 Burkholderia mallei
1.3.1.9 enoyl-ACP reductase
-
 Burkholderia mallei
1.3.1.9 FabV
-
 Burkholderia mallei
1.3.1.9 More the enzyme is a member of the short chain dehydrogenase/reductase superfamily Burkholderia mallei

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.3.1.9 25
-
 assay at Burkholderia mallei

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.3.1.9 0.022
-
 (2E)-2-dodecenoyl-CoA mutant K244R, pH 7.9, 25°C Burkholderia mallei
1.3.1.9 0.08
-
 (2E)-2-dodecenoyl-CoA mutant Y235A, pH 7.9, 25°C Burkholderia mallei
1.3.1.9 0.1
-
 (2E)-2-dodecenoyl-CoA mutant Y235S, pH 7.9, 25°C Burkholderia mallei
1.3.1.9 0.18
-
 (2E)-2-dodecenoyl-CoA mutant K244A, pH 7.9, 25°C Burkholderia mallei
1.3.1.9 0.3
-
 (2E)-2-dodecenoyl-CoA mutant K245M, pH 7.9, 25°C Burkholderia mallei
1.3.1.9 28.8
-
 (2E)-2-dodecenoyl-CoA wild-type, pH 7.9, 25°C Burkholderia mallei

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.3.1.9 7.9
-
-
 Burkholderia mallei
1.3.1.9 7.9
-
 assay at Burkholderia mallei

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.3.1.9 6.6 8.5
-
 Burkholderia mallei

Cofactor

EC Number Cofactor Comment Organism Structure
1.3.1.9 NADH
-
 Burkholderia mallei

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.3.1.9 additional information
-
 additional information inhibition kinetics, overview Burkholderia mallei
1.3.1.9 0.0004
-
 triclosan versus 2-dodecenoyl-CoA, pH 7.8, 25°C Burkholderia mallei
1.3.1.9 0.0004
-
 triclosan substrate 2-dodecenoyl-CoA, pH 7.9, 25°C Burkholderia mallei

General Information

EC Number General Information Comment Organism
1.3.1.9 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 [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.3.1.9 0.011
-
 (2E)-2-dodecenoyl-CoA mutant K245M, pH 7.9, 25°C Burkholderia mallei
1.3.1.9 0.025
-
 (2E)-2-dodecenoyl-CoA mutant K244A, pH 7.9, 25°C Burkholderia mallei
1.3.1.9 0.028
-
 (2E)-2-dodecenoyl-CoA mutant K244R, pH 7.9, 25°C Burkholderia mallei
1.3.1.9 0.03
-
 (2E)-2-dodecenoyl-CoA mutant Y235A, pH 7.9, 25°C Burkholderia mallei
1.3.1.9 0.047
-
 (2E)-2-dodecenoyl-CoA mutant Y235S, pH 7.9, 25°C Burkholderia mallei
1.3.1.9 6.5
-
 (2E)-2-dodecenoyl-CoA wild-type, pH 7.9, 25°C Burkholderia mallei