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Literature summary extracted from

  • Bondoc, J.; Wolf, N.; Ndichuck, M.; Abad-Zapatero, C.; Movahedzadeh, F.
    Mutagenesis of threonine to serine in the active site of Mycobacterium tuberculosis fructose-1,6-bisphosphatase (Class II) retains partial enzyme activity (2017), Biotechnol. Rep., 15, 48-54 .
No PubMed abstract available

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.3.11 gene glpX, sequence comparisons of class II FBPases, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Mycobacterium tuberculosis

Protein Variants

EC Number Protein Variants Comment Organism
3.1.3.11 T84A site-directed mutagenesis, the codon ACC for Thr84 is replaced by GCA for alanine, the active site mutant shows fully abolished enzyme activity while retaining substrate binding affinity Mycobacterium tuberculosis
3.1.3.11 T84A mutantion fully abolishes enzyme activity while retaining substrate binding affinity Mycobacterium tuberculosis
3.1.3.11 T84S site-directed mutagenesis, the codon ACC for Thr84 is replaced by AGC for serine, the active site mutant retains some activity having a 10times reduction in Vmax and exhibit similar sensitivity to lithium when compared to the wild-type enzyme Mycobacterium tuberculosis
3.1.3.11 T84S mutantion retains some activity having a 10 times reduction in Vmax and exhibits similar sensitivity to lithium when compared to the wild-type enzyme. Homology modeling using the Escherichia coli enzyme structure suggests that the replacement of the critical nucleophile OH- in the Thr84 residue of the wild-type enzyme by Ser84 results in subtle alterations of the position and orientation that reduces the catalytic efficiency. This mutant can be used to trap reaction intermediates, through crystallographic methods, facilitating the design of potent inhibitors via structure-based drug design Mycobacterium tuberculosis

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.1.3.11 Li+
-
 Mycobacterium tuberculosis
3.1.3.11 lithium
-
 Mycobacterium tuberculosis
3.1.3.11 additional information enzyme mutant T84S can be used to trap reaction intermediates, through crystallographic methods, facilitating the design of potent inhibitors via structure-based drug design Mycobacterium tuberculosis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1.3.11 additional information
-
 additional information kinetics Mycobacterium tuberculosis
3.1.3.11 0.0037
-
 D-fructose 1,6-bisphosphate pH 8.0, 22°C, recombinant wild-type enzyme Mycobacterium tuberculosis
3.1.3.11 0.0057
-
 D-fructose 1,6-bisphosphate pH 8.0, 22°C, recombinant mutant T84S Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.1.3.11 D-fructose 1,6-bisphosphate + H2O Mycobacterium tuberculosis
-
 D-fructose 6-phosphate + phosphate
-
 ?
3.1.3.11 D-fructose 1,6-bisphosphate + H2O Mycobacterium tuberculosis H37Rv
-
 D-fructose 6-phosphate + phosphate
-
 ?
3.1.3.11 D-fructose 1,6-bisphosphate + H2O Mycobacterium tuberculosis ATCC 25618
-
 D-fructose 6-phosphate + phosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.1.3.11 Mycobacterium tuberculosis P9WN21
-
-
3.1.3.11 Mycobacterium tuberculosis ATCC 25618 P9WN21
-
-
3.1.3.11 Mycobacterium tuberculosis H37Rv P9WN21
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.3.11
-
 Mycobacterium tuberculosis
3.1.3.11 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, and desalting gel filtration Mycobacterium tuberculosis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.3.11 D-fructose 1,6-bisphosphate + H2O
-
 Mycobacterium tuberculosis D-fructose 6-phosphate + phosphate
-
 ?
3.1.3.11 D-fructose 1,6-bisphosphate + H2O
-
 Mycobacterium tuberculosis H37Rv D-fructose 6-phosphate + phosphate
-
 ?
3.1.3.11 D-fructose 1,6-bisphosphate + H2O
-
 Mycobacterium tuberculosis ATCC 25618 D-fructose 6-phosphate + phosphate
-
 ?
3.1.3.11 additional information substrate binding analysis of wild-type and mutant enzymes by surface plasmon resonance method, substrate affinities, overview Mycobacterium tuberculosis ?
-
 ?
3.1.3.11 additional information substrate binding analysis of wild-type and mutant enzymes by surface plasmon resonance method, substrate affinities, overview Mycobacterium tuberculosis H37Rv ?
-
 ?
3.1.3.11 additional information substrate binding analysis of wild-type and mutant enzymes by surface plasmon resonance method, substrate affinities, overview Mycobacterium tuberculosis ATCC 25618 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.1.3.11 ? x * 36584, recombinant mutant T84A, mass spectrometry and sequence calculation, x * 36599, recombinant mutant T84S, mass spectrometry and sequence calculation Mycobacterium tuberculosis

Synonyms

EC Number Synonyms Comment Organism
3.1.3.11 class II fructose-1,6-bisphosphatase
-
 Mycobacterium tuberculosis
3.1.3.11 fructose-1,6-bisphosphatase
-
 Mycobacterium tuberculosis
3.1.3.11 fructose-1,6-bisphosphatase class II
-
 Mycobacterium tuberculosis
3.1.3.11 GlpX
-
 Mycobacterium tuberculosis
3.1.3.11 MtFBPaseII
-
 Mycobacterium tuberculosis
3.1.3.11 MtFPBase
-
 Mycobacterium tuberculosis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.1.3.11 22
-
 assay at room temperature Mycobacterium tuberculosis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.1.3.11 0.22
-
 D-fructose 1,6-bisphosphate pH 8.0, 22°C, recombinant mutant T84S Mycobacterium tuberculosis
3.1.3.11 2.1
-
 D-fructose 1,6-bisphosphate pH 8.0, 22°C, recombinant wild-type enzyme Mycobacterium tuberculosis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.1.3.11 8
-
 assay at Mycobacterium tuberculosis

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
3.1.3.11 0.25
-
 pH 8.0, 22°C, recombinant wild-type enzyme Mycobacterium tuberculosis Li+
3.1.3.11 0.28
-
 pH 8.0, 22°C, recombinant mutant T84S Mycobacterium tuberculosis Li+

General Information

EC Number General Information Comment Organism
3.1.3.11 malfunction homology modeling using the Escherichia coli enzyme structure suggests that the replacement of the critical nucleophile OH- in the Thr84 residue of the wild-type MtFBPase by Ser84 results in subtle alterations of the position and orientation that reduce the catalytic efficiency Mycobacterium tuberculosis
3.1.3.11 metabolism essential enzyme for pathogenesis Mycobacterium tuberculosis
3.1.3.11 additional information Thr84 in MtFBPaseII protein (Thr90 in Escherichia coli) is a conserved residue in the active site as shown in other FBPases Mycobacterium tuberculosis
3.1.3.11 physiological function class II fructose-1,6-bisphosphatase enzyme in Mycobacterium tuberculosis is an essential enzyme for pathogenesis Mycobacterium tuberculosis