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Literature summary extracted from

  • Lindner, S.N.; Knebel, S:; Wesseling,H.; Schoberth, S.M.; Wendisch, V.F.
    Exopolyphosphatases PPX1 and PPX2 from Corynebacterium glutamicum[down-pointing small open triangle] (2009), Appl. Environ. Microbiol., 75, 3161-3170.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.6.1.11 additional information CaCl2, BaCl2, CuSO4, and SnCl2 do not show any activating effect on PPX2 Corynebacterium glutamicum

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.6.1.11 expression in Escherichia coli, overexpression of ppx2 gene in Corynebacterium glutamicum results in higher exopolyphosphatase activities in crude extracts and deletion of the gene with lower activities than those of the wild-type strain Corynebacterium glutamicum
3.6.1.11 overexpression of ppx1 gene in Corynebacterium glutamicum results in higher exopolyphosphatase activities in crude extracts and deletion of the gene with lower activities than those of the wild-type strain Corynebacterium glutamicum

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.6.1.11 ADP
-
 Corynebacterium glutamicum
3.6.1.11 ATP
-
 Corynebacterium glutamicum
3.6.1.11 Ca2+ PPX2 is inhibited by millimolar concentrations Corynebacterium glutamicum
3.6.1.11 CaCl2 3.8 mM CaCl2 leads to half-maximal inhibition of His-tagged PPX2 Corynebacterium glutamicum
3.6.1.11 dATP
-
 Corynebacterium glutamicum
3.6.1.11 dGTP
-
 Corynebacterium glutamicum
3.6.1.11 GTP
-
 Corynebacterium glutamicum
3.6.1.11 ITP
-
 Corynebacterium glutamicum
3.6.1.11 Mg2+ PPX2 is inhibited by millimolar concentrations, activity is reduced 2fold at 10 mM MnCl2 Corynebacterium glutamicum
3.6.1.11 Mn2+ PPX2 is inhibited by millimolar concentrations, activity is reduced 2fold at 3.5 mM MnCl2 Corynebacterium glutamicum
3.6.1.11 Pyrophosphate 8.2 mM pyrophosphate leads to half-maximal inhibition of His-tagged PPX2 Corynebacterium glutamicum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.6.1.11 0.04
-
 Triphosphate in 50 mM PIPES, pH 6.8 containing 25 mM KCl and 2 mM MgCl2 Corynebacterium glutamicum
3.6.1.11 0.11
-
 Tetraphosphate in 50 mM PIPES, pH 6.8 containing 25 mM KCl and 2 mM MgCl2 Corynebacterium glutamicum

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.6.1.11 Fe2+ enzyme is stimulated 0.26fold at a concentration of 2 mM Corynebacterium glutamicum
3.6.1.11 K+ PPX2 activity is increased, 25 mM KCl resulting in a 3fold increase in the specific activity Corynebacterium glutamicum
3.6.1.11 Mg2+ enzyme requires Mg2+ cations but is inhibited by higher concentrations, Mg2+ shows the highest stimulation at 2 mM Corynebacterium glutamicum
3.6.1.11 Mn2+ enzyme is stimulated 0.86fold at a concentration of 2 mM Corynebacterium glutamicum
3.6.1.11 Zn2+ enzyme is stimulated 0.11fold at a concentration of 2 mM Corynebacterium glutamicum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.6.1.11 (phosphate)n + H2O Corynebacterium glutamicum degradation of polyphosphate (phosphate)n-1 + phosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.6.1.11 Corynebacterium glutamicum Q8NRR8
-
-
3.6.1.11 Corynebacterium glutamicum Q8NT99
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.6.1.11 recombinant ppx2 enzyme is purified by Ni-nitrilotriacetic acid chromatography Corynebacterium glutamicum

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.6.1.11 additional information
-
 when various short- to medium-chain PolyPs are tested, His-tagged PPX2 is most active with polyphosphates of 3-20 phosphate residues Corynebacterium glutamicum
3.6.1.11 0.04
-
 analysis of the constructed deletion mutant ppx2 reveal reduced exopolyphosphatase activity, samples containing 50 mM PIPES, pH 6.8, 25 mM KCl, and 2 mM MgCl2 Corynebacterium glutamicum
3.6.1.11 0.08
-
 analysis of the constructed deletion mutant ppx1 reveal reduced exopolyphosphatase activity, samples containing 50 mM PIPES, pH 6.8, 25 mM KCl, and 2 mM MgCl2 Corynebacterium glutamicum
3.6.1.11 0.12
-
 wild type, samples containing 50 mM PIPES, pH 6.8, 25 mM KCl, and 2 mM MgCl2 Corynebacterium glutamicum
3.6.1.11 0.43
-
 overexpression of ppx1 increase the specific activity of exopolyphosphatase by 4fold compared to that of the empty vector control Corynebacterium glutamicum
3.6.1.11 0.8
-
 crude cell extracts of wild type strain pVWEx1-ppx2 show 6fold higher exopolyphosphatase activity than those of wild type pVWEx1 Corynebacterium glutamicum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.6.1.11 (phosphate)n + H2O degradation of polyphosphate Corynebacterium glutamicum (phosphate)n-1 + phosphate
-
 ?
3.6.1.11 additional information PPX2 is active with short-chain polyphosphates, even accepting diphosphate Corynebacterium glutamicum ?
-
 ?
3.6.1.11 tetraphosphate + H2O
-
 Corynebacterium glutamicum triphosphate + phosphate
-
 ?
3.6.1.11 triphosphate + H2O
-
 Corynebacterium glutamicum diphosphate + phosphate
-
 ?

Subunits

EC Number Subunits Comment Organism
3.6.1.11 monomer determined by gel filtration Corynebacterium glutamicum

Synonyms

EC Number Synonyms Comment Organism
3.6.1.11 exopolyphosphatase
-
 Corynebacterium glutamicum
3.6.1.11 PPX1
-
 Corynebacterium glutamicum
3.6.1.11 PPX2
-
 Corynebacterium glutamicum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.6.1.11 30
-
-
 Corynebacterium glutamicum

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.6.1.11 25 40 at pH 6.8, 2 mM MgCl2 and 25 mM KCl, no significant loss of activity is observed after 60 min Corynebacterium glutamicum
3.6.1.11 45
-
 after preheating at 45°C for 30 and 60 min, 70% and 40% of the PPX2 activity remains Corynebacterium glutamicum
3.6.1.11 50
-
 after preheating at 50°C, His-tagged PPX2 loses its activity quickly, after incubation for 60 min, no activity remains Corynebacterium glutamicum
3.6.1.11 60
-
 complete inactivation of His-tagged PPX2 within 5 min Corynebacterium glutamicum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.6.1.11 0.6
-
 Triphosphate in 50 mM PIPES, pH 6.8 containing 25 mM KCl and 2 mM MgCl2 Corynebacterium glutamicum
3.6.1.11 0.9
-
 Tetraphosphate in 50 mM PIPES, pH 6.8 containing 25 mM KCl and 2 mM MgCl2 Corynebacterium glutamicum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.6.1.11 6.8
-
 highest activity of recombinant His-tagged PPX2 is reached in 50 mM PIPES buffer Corynebacterium glutamicum

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.6.1.11 0.4
-
 dGTP
-
 Corynebacterium glutamicum
3.6.1.11 0.6
-
 ADP
-
 Corynebacterium glutamicum
3.6.1.11 0.6
-
 dATP
-
 Corynebacterium glutamicum
3.6.1.11 0.8
-
 GTP
-
 Corynebacterium glutamicum
3.6.1.11 0.8
-
 ITP
-
 Corynebacterium glutamicum
3.6.1.11 2.6
-
 ATP
-
 Corynebacterium glutamicum