Literature summary for 2.3.1.180 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.

Search for more literature for 2.3.1.180

Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment)	Organism
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

Crystallization (Comment)

Organism

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

Protein Variants	Comment	Organism
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
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
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
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
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
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
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
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
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

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
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	-	?
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

Organism	UniProt	Comment	Textmining
Mycobacterium tuberculosis	P9WNG3	gene fabH	-
Mycobacterium tuberculosis H37Rv	P9WNG3	gene fabH	-

Purification (Commentary)

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

Reaction

Reaction	Comment	Organism	Reaction ID
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

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
acetyl-CoA + malonyl-[acyl-carrier protein]	-	Mycobacterium tuberculosis	acetoacetyl-[acyl-carrier protein] + CoA + CO2	-	?
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	-	?
acetyl-CoA + malonyl-[acyl-carrier protein]	-	Mycobacterium tuberculosis H37Rv	acetoacetyl-[acyl-carrier protein] + CoA + CO2	-	?
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	-	?
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	?	-	?
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

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

Temperature Optimum [°C]

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

pH Optimum

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