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Literature summary extracted from

  • Brown, A.K.; Sridharan, S.; Kremer, L.; Lindenberg, S.; Dover, L.G.; Sacchettini, J.C.; Besra, G.S.
    Probing the mechanism of the Mycobacterium tuberculosis beta-ketoacyl-ACP synthase III mtFabH: Factors influencing catalysis and substrate specificity (2005), J. Biol. Chem., 280, 32539-32547.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.1.180 overexpression of His-tagged wild-type and mutant enzymes in strain C41(DE3) Mycobacterium tuberculosis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.3.1.180 purified recombinant mutant enzymes R46A/R161A and W42A/R163A, 0.003 ml R46A/R161A solution containing 20 mg/ml protein, 20 mM Tris-HCl, pH 7.5, 0.5 M NaCl, 2 mM 2-mercaptoethanol, 10% glycerol, mixed with 0.001 ml of crystallization solution containing 0.1 M HEPES, pH 7.5, 10% v/v isopropyl alcohol, 20% w/v PEG 4000, for the W42A/R163A solution 0.002 ml of protein solution is mixed with 0.001 ml of crystallization solution containing 0.2 M ammonium sulfate, and 30% w/v PEG 8000, X-ray diffraction structure determination and analysis at 2.0-2.65 A resolution Mycobacterium tuberculosis

Protein Variants

EC Number Protein Variants Comment Organism
2.3.1.41 R161A 69.6% of wild-type activity Mycobacterium tuberculosis
2.3.1.41 R46A 7.3% of wild-type activity Mycobacterium tuberculosis
2.3.1.41 R46A/R161A 0.3% of wild-type activity Mycobacterium tuberculosis
2.3.1.41 T97F no enzymic activity Mycobacterium tuberculosis
2.3.1.41 W42A 21.5% of wild-type activity Mycobacterium tuberculosis
2.3.1.41 W42A/R161A 0.2% of wild-type activity Mycobacterium tuberculosis
2.3.1.180 C122A site-directed mutagenesis, the mutant shows 0.25% of wild-type enzyme condensation activity, and 2.4% of wild-type malonyl-ACP decarboxylation activity Mycobacterium tuberculosis
2.3.1.180 H258A site-directed mutagenesis, the mutant shows 0.46% of wild-type enzyme condensation activity, and 0.68% of wild-type malonyl-ACP decarboxylation activity Mycobacterium tuberculosis
2.3.1.180 N289A site-directed mutagenesis, the mutant shows 0.51% of wild-type enzyme condensation activity, and 2.24% of wild-type malonyl-ACP decarboxylation activity Mycobacterium tuberculosis
2.3.1.180 R161A site-directed mutagenesis, the mutant shows 69.7% of wild-type enzyme condensation activity, and 8.7% of wild-type malonyl-ACP decarboxylation activity Mycobacterium tuberculosis
2.3.1.180 R46A site-directed mutagenesis, the mutant shows 7.3% of wild-type enzyme condensation activity, and 0.79% of wild-type malonyl-ACP decarboxylation activity Mycobacterium tuberculosis
2.3.1.180 R46A/R161A site-directed mutagenesis, the mutant shows 0.31% of wild-type enzyme condensation activity, and 5.1% of wild-type malonyl-ACP decarboxylation activity Mycobacterium tuberculosis
2.3.1.180 T97F site-directed mutagenesis, the mutant shows 0.47% of wild-type enzyme condensation activity, and 0.84% of wild-type malonyl-ACP decarboxylation activity Mycobacterium tuberculosis
2.3.1.180 W42A site-directed mutagenesis, the mutant shows 21.6% of wild-type enzyme condensation activity, and 30.1% of wild-type malonyl-ACP decarboxylation activity Mycobacterium tuberculosis
2.3.1.180 W42A/R161A site-directed mutagenesis, the mutant shows 0.24% of wild-type enzyme condensation activity, and 9.7% of wild-type malonyl-ACP decarboxylation activity Mycobacterium tuberculosis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.1.41 0.0023
-
 dodecanoyl-[acyl-carrier protein] pH 7.0, 37°C Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.3.1.180 acetyl-CoA + malonyl-[acyl-carrier protein] Mycobacterium tuberculosis condensation of fatty acid synthase I-derived acyl-CoAs with malonyl-ACPs to initiates fatty acid synthase II-catalyzed fatty acid elongation, the enzyme links the two fatty acid synthase complexes acetoacetyl-[acyl-carrier protein] + CoA + CO2
-
 ?
2.3.1.180 acetyl-CoA + malonyl-[acyl-carrier protein] Mycobacterium tuberculosis H37Rv condensation of fatty acid synthase I-derived acyl-CoAs with malonyl-ACPs to initiates fatty acid synthase II-catalyzed fatty acid elongation, the enzyme links the two fatty acid synthase complexes acetoacetyl-[acyl-carrier protein] + CoA + CO2
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.3.1.41 Mycobacterium tuberculosis
-
-
-
2.3.1.180 Mycobacterium tuberculosis P9WNG3 gene fabH
-
2.3.1.180 Mycobacterium tuberculosis H37Rv P9WNG3 gene fabH
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.1.180 recombinant His-tagged wild-type and mutant enzymes from strain C41(DE3) by nickel affinity chromatography Mycobacterium tuberculosis

Reaction

EC Number Reaction Comment Organism Reaction ID
2.3.1.180 acetyl-CoA + a malonyl-[acyl-carrier protein] = an acetoacetyl-[acyl-carrier protein] + CoA + CO2 active site structure and catalytic reaction mechanism, T97, R46, W42, C122, R161, H258, and N289 are important for activity, structure-function relationship Mycobacterium tuberculosis
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.3.1.41 0.0017
-
 substrate hexanoyl-[acyl-carrier protein], pH 7.0, 37°C Mycobacterium tuberculosis
2.3.1.41 0.0055
-
 substrate octanoyl-[acyl-carrier protein], pH 7.0, 37°C Mycobacterium tuberculosis
2.3.1.41 0.0062
-
 substrate decanoyl-[acyl-carrier protein], pH 7.0, 37°C Mycobacterium tuberculosis
2.3.1.41 0.0072
-
 substrate dodecanoyl-[acyl-carrier protein], pH 7.0, 37°C Mycobacterium tuberculosis
2.3.1.41 0.009
-
 substrate eicosanoyl-[acyl-carrier-protein], pH 7.0, 37°C Mycobacterium tuberculosis
2.3.1.41 0.0093
-
 substrate palmitoyl-[acyl-carrier protein], pH 7.0, 37°C Mycobacterium tuberculosis
2.3.1.41 0.0095
-
 substrate myristoyl-[acyl-carrier protein], pH 7.0, 37°C Mycobacterium tuberculosis
2.3.1.41 0.0104
-
 substrate octadecanoyl-[acyl-carrier-protein], pH 7.0, 37°C Mycobacterium tuberculosis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.1.41 decanoyl-[acyl-carrier protein] + malonyl-[acyl-carrier protein]
-
 Mycobacterium tuberculosis dodecanoyl-[acyl-carrier protein] + CO2 + [acyl-carrier protein]
-
 ?
2.3.1.41 dodecanoyl-[acyl-carrier protein] + malonyl-[acyl-carrier protein]
-
 Mycobacterium tuberculosis 3-oxotetradecanoyl-[acyl-carrier protein] + CO2 + [acyl-carrier protein]
-
 ?
2.3.1.41 eicosanoyl-[acyl-carrier protein] + malonyl-[acyl-carrier protein]
-
 Mycobacterium tuberculosis 3-oxodocosanoyl-[acyl-carrier protein] + CO2 + [acyl-carrier protein]
-
 ?
2.3.1.41 hexanoyl-[acyl-carrier protein] + malonyl-[acyl-carrier protein]
-
 Mycobacterium tuberculosis 3-oxooctanoyl-[acyl-carrier protein] + CO2 + [acyl-carrier protein]
-
 ?
2.3.1.41 additional information enzyme is able to use Escherichia coli acyl-carrier protein. Enzyme extends longer, physiologically relevant acyl-CoA primers when paired with its native acyl-carrier protein than with Escherichia coli acyl-carrier protein Mycobacterium tuberculosis ?
-
 ?
2.3.1.41 myristoyl-[acyl-carrier protein] + malonyl-[acyl-carrier protein]
-
 Mycobacterium tuberculosis 3-oxohexadecanoyl-[acyl-carrier protein] + CO2 + [acyl-carrier protein]
-
 ?
2.3.1.41 octadecanoyl-[acyl-carrier protein] + malonyl-[acyl-carrier protein]
-
 Mycobacterium tuberculosis 3-oxoeicosanoyl-[acyl-carrier protein] + CO2 + [acyl-carrier protein]
-
 ?
2.3.1.41 octanoyl-[acyl-carrier protein] + malonyl-[acyl-carrier protein]
-
 Mycobacterium tuberculosis 3-oxodecanoyl-[acyl-carrier protein] + CO2 + [acyl-carrier protein]
-
 ?
2.3.1.41 palmitoyl-[acyl-carrier protein] + malonyl-[acyl-carrier protein]
-
 Mycobacterium tuberculosis 3-oxooctadecanoyl-[acyl-carrier protein] + CO2 + [acyl-carrier protein]
-
 ?
2.3.1.180 acetyl-CoA + malonyl-[acyl-carrier protein]
-
 Mycobacterium tuberculosis acetoacetyl-[acyl-carrier protein] + CoA + CO2
-
 ?
2.3.1.180 acetyl-CoA + malonyl-[acyl-carrier protein] condensation of fatty acid synthase I-derived acyl-CoAs with malonyl-ACPs to initiates fatty acid synthase II-catalyzed fatty acid elongation, the enzyme links the two fatty acid synthase complexes Mycobacterium tuberculosis acetoacetyl-[acyl-carrier protein] + CoA + CO2
-
 ?
2.3.1.180 acetyl-CoA + malonyl-[acyl-carrier protein]
-
 Mycobacterium tuberculosis H37Rv acetoacetyl-[acyl-carrier protein] + CoA + CO2
-
 ?
2.3.1.180 acetyl-CoA + malonyl-[acyl-carrier protein] condensation of fatty acid synthase I-derived acyl-CoAs with malonyl-ACPs to initiates fatty acid synthase II-catalyzed fatty acid elongation, the enzyme links the two fatty acid synthase complexes Mycobacterium tuberculosis H37Rv acetoacetyl-[acyl-carrier protein] + CoA + CO2
-
 ?
2.3.1.180 additional information specificity of the enzyme for longer acyl-CoA chains, overview, the enzyme shows also acyl-CoA-[acyl-carrier-protein] transacylase activity Mycobacterium tuberculosis ?
-
 ?
2.3.1.180 additional information specificity of the enzyme for longer acyl-CoA chains, overview, the enzyme shows also acyl-CoA-[acyl-carrier-protein] transacylase activity Mycobacterium tuberculosis H37Rv ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.3.1.180 beta-ketoacyl-acyl carrier protein synthase III
-
 Mycobacterium tuberculosis
2.3.1.180 FabH
-
 Mycobacterium tuberculosis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.3.1.180 37
-
 assay at Mycobacterium tuberculosis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.1.180 7
-
 assay at, condensation activity Mycobacterium tuberculosis
2.3.1.180 8
-
 assay at, decarboxylation activity Mycobacterium tuberculosis