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Literature summary for 2.1.2.11 extracted from

  • Sugantino, M.; Zheng, R.; Yu, M.; Blanchard, J.S.
    Mycobacterium tuberculosis ketopantoate hydroxymethyltransferase: Tetrahydrofolate-independent hydroxymethyltransferase and enolization reactions with alpha-keto acids (2003), Biochemistry, 42, 191-199.
    View publication on PubMed

Application

Application Comment Organism
pharmacology enzyme might be an attractive target for inhibitor design Mycobacterium tuberculosis

Cloned(Commentary)

Cloned (Comment) Organism
panB gene encoding enzyme is cloned and overexpressed in Escherichia coli BL21 (DE3) Mycobacterium tuberculosis

Inhibitors

Inhibitors Comment Organism Structure
additional information not inhibited by methylenetetrahydrofolate, i.e. equimolar formaldehyde and tetrahydrofolate, at concentrations up to 2 mM Mycobacterium tuberculosis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information
-
Mycobacterium tuberculosis
0.24
-
3-methyl-2-oxobutanoate
-
Mycobacterium tuberculosis
0.82
-
5,10-methylenetetrahydrofolate
-
Mycobacterium tuberculosis

Localization

Localization Comment Organism GeneOntology No. Textmining
soluble
-
Mycobacterium tuberculosis
-
-

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ substrate enolization is divalent metal-dependent with a preference for metal ions in decreasing order: Mg2+, Zn2+, Co2+, Ni2+, Ca2+ Mycobacterium tuberculosis
Ca2+ Km: 0.27 mM, the reaction is divalent metal-dependent with descending order of preference: Mg2+, Zn2+, Co2+, Ni2+, Ca2+ Mycobacterium tuberculosis
Co2+ substrate enolization is divalent metal-dependent with a preference for metal ions in decreasing order: Mg2+, Zn2+, Co2+, Ni2+, Ca2+ Mycobacterium tuberculosis
Co2+ Km: 0.33 mM, the reaction is divalent metal-dependent with descending order of preference: Mg2+, Zn2+, Co2+, Ni2+, Ca2+ Mycobacterium tuberculosis
Mg2+ substrate enolization is divalent metal-dependent with a preference for metal ions in decreasing order: Mg2+, Zn2+, Co2+, Ni2+, Ca2+ Mycobacterium tuberculosis
Mg2+ Km: 0.61 mM, the reaction is divalent metal-dependent with descending order of preference: Mg2+, Zn2+, Co2+, Ni2+, Ca2+ Mycobacterium tuberculosis
additional information metalloenzyme, inactive in absence of divalent metals, enzyme binds metal ions that assist in the polarization of the carbonyl group and stabilize the enolate anion Mycobacterium tuberculosis
Ni2+ substrate enolization is divalent metal-dependent with a preference for metal ions in decreasing order: Mg2+, Zn2+, Co2+, Ni2+, Ca2+ Mycobacterium tuberculosis
Ni2+ Km: 0.45 mM, the reaction is divalent metal-dependent with descending order of preference: Mg2+, Zn2+, Co2+, Ni2+, Ca2+ Mycobacterium tuberculosis
Zn2+ substrate enolization is divalent metal-dependent with a preference for metal ions in decreasing order: Mg2+, Zn2+, Co2+, Ni2+, Ca2+ Mycobacterium tuberculosis
Zn2+ Km: 0.08 mM, the reaction is divalent metal-dependent with descending order of preference: Mg2+, Zn2+, Co2+, Ni2+, Ca2+ Mycobacterium tuberculosis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
29000
-
recombinant enzyme expressed in Escherichia coli BL21 (DE3): 4 * 29000, SDS-PAGE, 4 * 29202, electrospray ionization/mass spectrometry, 4 * 29366, calculated from the amino acid sequence Mycobacterium tuberculosis
29202
-
recombinant enzyme expressed in Escherichia coli BL21 (DE3): 4 * 29000, SDS-PAGE, 4 * 29202, electrospray ionization/mass spectrometry, 4 * 29366, calculated from the amino acid sequence Mycobacterium tuberculosis
29366
-
recombinant enzyme expressed in Escherichia coli BL21 (DE3): 4 * 29000, SDS-PAGE, 4 * 29202, electrospray ionization/mass spectrometry, 4 * 29366, calculated from the amino acid sequence Mycobacterium tuberculosis
120000
-
gel filtration Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
5,10-methylenetetrahydrofolate + 3-methyl-2-oxobutanoate Mycobacterium tuberculosis first committed step in pantothenate biosynthesis tetrahydrofolate + 2-dehydropantoate
-
?
5,10-methylenetetrahydrofolate + 3-methyl-2-oxobutanoate Mycobacterium tuberculosis enzyme may be the rate-limiting reaction in pantothenate biosynthesis tetrahydrofolate + 2-dehydropantoate
-
?

Organism

Organism UniProt Comment Textmining
Mycobacterium tuberculosis
-
-
-

Purification (Commentary)

Purification (Comment) Organism
purification of recombinant enzyme expressed in Escherichia coli BL21 (DE3) Mycobacterium tuberculosis

Reaction

Reaction Comment Organism Reaction ID
5,10-methylenetetrahydrofolate + 3-methyl-2-oxobutanoate + H2O = tetrahydrofolate + 2-dehydropantoate stereochemistry Mycobacterium tuberculosis
5,10-methylenetetrahydrofolate + 3-methyl-2-oxobutanoate + H2O = tetrahydrofolate + 2-dehydropantoate sequential kinetic mechanism, chemical mechanism Mycobacterium tuberculosis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5,10-methylenetetrahydrofolate + 2-oxo-4-methylthiobutyrate about 50% of the activity with 3-methyl-2-oxobutanoate Mycobacterium tuberculosis tetrahydrofolate + ?
-
?
5,10-methylenetetrahydrofolate + 2-oxobutyrate about 50% of the activity with 3-methyl-2-oxobutanoate Mycobacterium tuberculosis tetrahydrofolate + ?
-
?
5,10-methylenetetrahydrofolate + 2-oxopentanoate about 30% of the activity with 3-methyl-2-oxobutanoate Mycobacterium tuberculosis tetrahydrofolate + ?
-
?
5,10-methylenetetrahydrofolate + 3-methyl-2-oxobutanoate
-
Mycobacterium tuberculosis tetrahydrofolate + 2-dehydropantoate
-
?
5,10-methylenetetrahydrofolate + 3-methyl-2-oxobutanoate first committed step in pantothenate biosynthesis Mycobacterium tuberculosis tetrahydrofolate + 2-dehydropantoate
-
?
5,10-methylenetetrahydrofolate + 3-methyl-2-oxobutanoate enzyme may be the rate-limiting reaction in pantothenate biosynthesis Mycobacterium tuberculosis tetrahydrofolate + 2-dehydropantoate
-
?
5,10-methylenetetrahydrofolate + 3-methyl-2-oxopentanoate about 65% of the activity with 3-methyl-2-oxobutanoate Mycobacterium tuberculosis tetrahydrofolate + ?
-
?
5,10-methylenetetrahydrofolate + pyruvate about 20% of the activity with 3-methyl-2-oxobutanoate Mycobacterium tuberculosis tetrahydrofolate + ?
-
?
formaldehyde + 3-methyl-2-oxobutanoate enzyme catalyzes methylenetetrahydrofolate-independent hydroxymethyltransferase reaction between free formaldehyde and alpha-ketoisovalerate, formaldehyde is unlikely to be the natural substrate Mycobacterium tuberculosis 2-dehydropantoate
-
?
additional information enzyme catalyzes deuterium exchange in the methylenetetrahydrofolate-independent enolization of alpha-ketoisovalerate or other alpha-keto acids with decreasing efficiency: alpha-ketoisovalerate, alpha-ketobutyrate, alpha-ketovalerate, pyruvate, alpha-ketomethylthiobutyrate, alpha-ketoisocaproate, stereochemistry, first step in the reaction leading to ketopantoate is the enolization of alpha-ketoisovalerate to form the stabilized carbanion Mycobacterium tuberculosis ?
-
?

Subunits

Subunits Comment Organism
tetramer recombinant enzyme expressed in Escherichia coli BL21 (DE3): 4 * 29000, SDS-PAGE, 4 * 29202, electrospray ionization/mass spectrometry, 4 * 29366, calculated from the amino acid sequence Mycobacterium tuberculosis

Synonyms

Synonyms Comment Organism
ketopantoate hydroxymethyltransferase
-
Mycobacterium tuberculosis
PanB
-
Mycobacterium tuberculosis

Temperature Optimum [°C]

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

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.78
-
5,10-methylenetetrahydrofolate
-
Mycobacterium tuberculosis
0.78
-
3-methyl-2-oxobutanoate
-
Mycobacterium tuberculosis
0.783
-
3-methyl-2-oxobutanoate
-
Mycobacterium tuberculosis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7 7.5
-
Mycobacterium tuberculosis
7 7.5 rate of substrate enolization is pH-dependent with optimal activity in the range of Mycobacterium tuberculosis

pH Stability

pH Stability pH Stability Maximum Comment Organism
5.5
-
loss of proper protein folding at pH values lower than 5.5 Mycobacterium tuberculosis

Cofactor

Cofactor Comment Organism Structure
5,10-methylenetetrahydrofolate 5,10-methylenetetrahydrofolate as cofactor Mycobacterium tuberculosis
tetrahydrofolate
-
Mycobacterium tuberculosis