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Literature summary for 2.7.4.1 extracted from

  • Lindner, S.N.; Vidaurre, D.; Willbold, S.; Schoberth, S.M.; Wendisch, V.F.
    NCgl2620 encodes a class II polyphosphate kinase in Corynebacterium glutamicum (2007), Appl. Environ. Microbiol., 73, 5026-5033.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.7.4.1

Activating Compound

Activating Compound Comment Organism Structure
(NH4)2SO4 optimal at 80 mM Corynebacterium glutamicum

Cloned(Commentary)

Cloned (Comment) Organism
genes NCgl0880 and NCgl2620 or pkk2A and pkk2B, expression of wild-type and mutant PKK2B isozymes in Escherichia coli strain Bl21(DE3), subcloning in Escherichia coli strain DH5alpha Corynebacterium glutamicum
overexpression in Escherichia coli Corynebacterium glutamicum

Protein Variants

Protein Variants Comment Organism
additional information ppk2A and ppk2b knockout organisms, ppk2b has PPK activity Corynebacterium glutamicum
additional information deletion of ppk2B decreases PPK activity and cellular polyphosphate content, while overexpression of ppk2B increases both PPK activity and cellular polyphosphate content. Neither deletion nor overexpression of ppk2A changes specific activity of PPK or cellular polyphosphate content significantly. The ppk2B deletion mutant, which accumulated very little polyphopshate and grows like the wild-type under phosphate-sufficient conditions, shows a growth defect under phosphate-limiting conditions, phenotype, overview Corynebacterium glutamicum

Inhibitors

Inhibitors Comment Organism Structure
AMP
-
 Corynebacterium glutamicum
GMP
-
 Corynebacterium glutamicum
IMP
-
 Corynebacterium glutamicum
additional information no inhibition by UMP and CMP Corynebacterium glutamicum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.14
-
 GTP
-
 Corynebacterium glutamicum
0.14
-
 GTP pH 7.2, 30°C, recombinant wild-type enzyme Corynebacterium glutamicum
0.17
-
 ATP
-
 Corynebacterium glutamicum
0.17
-
 ATP pH 7.2, 30°C, recombinant wild-type enzyme Corynebacterium glutamicum
1
-
 ITP
-
 Corynebacterium glutamicum
1
-
 ITP pH 7.2, 30°C, recombinant wild-type enzyme Corynebacterium glutamicum
15
-
 (Phosphate)n
-
 Corynebacterium glutamicum
15
-
 Polyphosphate pH 7.2, 30°C, recombinant wild-type enzyme Corynebacterium glutamicum

Localization

Localization Comment Organism GeneOntology No. Textmining
cytosol
-
 Corynebacterium glutamicum 5829
-

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ optimal at 10 mM Corynebacterium glutamicum
Mg2+ optimal at 10 mM, activates Corynebacterium glutamicum
Mn2+ optimal at 10 mM Corynebacterium glutamicum
Mn2+ optimal at 10 mM, activates highly Corynebacterium glutamicum

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
151400
-
 gel filtration Corynebacterium glutamicum

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + (phosphate)n Corynebacterium glutamicum PPK2B forms polyphopshate with an average chain length of about 125 ADP + (phosphate)n+1
-
 r
ATP + (phosphate)n Corynebacterium glutamicum MH20-22B PPK2B forms polyphopshate with an average chain length of about 125 ADP + (phosphate)n+1
-
 r
additional information Corynebacterium glutamicum accumulation of high levels of cytosolic phosphorus in the form of polyphosphate involving classII polyphosphate kinase isozymes, granular poly P, i.e. volutin, can make up to 37% of the internal cell volume, overview ?
-
 ?
additional information Corynebacterium glutamicum MH20-22B accumulation of high levels of cytosolic phosphorus in the form of polyphosphate involving classII polyphosphate kinase isozymes, granular poly P, i.e. volutin, can make up to 37% of the internal cell volume, overview ?
-
 ?

Organism

Organism UniProt Comment Textmining
Corynebacterium glutamicum
-
 MH20-22B
-
Corynebacterium glutamicum
-
 wild-type strain ATCC 13032 is auxotrophic for biotin, genes NCgl0880 and NCgl2620 encoding class II isozymes PPK2A and PPK2B, no class I polyphosphate kinase, PPK1, encoded by ppk1, in this bacterium
-
Corynebacterium glutamicum MH20-22B
-
 MH20-22B
-

Purification (Commentary)

Purification (Comment) Organism
of the overexpessed protein Corynebacterium glutamicum

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
 Corynebacterium glutamicum
14
-
 purified recombinant wild-type enzyme, polyphosphate synthesis with GTP Corynebacterium glutamicum
18
-
 purified recombinant wild-type enzyme, polyphosphate synthesis with ITP Corynebacterium glutamicum
31
-
 purified recombinant wild-type enzyme, polyphosphate synthesis with ATP Corynebacterium glutamicum
32
-
 purified recombinant wild-type enzyme, polyphosphate dephosphorylation Corynebacterium glutamicum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + (phosphate)n
-
 Corynebacterium glutamicum ADP + (phosphate)n+1
-
 r
ATP + (phosphate)n PPK2B forms polyphopshate with an average chain length of about 125 Corynebacterium glutamicum ADP + (phosphate)n+1
-
 r
ATP + (phosphate)n PPK2B forms polyphopshate with an average chain length of about 125 and prefers the polyphosphate synthesis reaction direction Corynebacterium glutamicum ADP + (phosphate)n+1 polyphosphate product chain length determination by NMR spectroscopy r
ATP + (phosphate)n
-
 Corynebacterium glutamicum MH20-22B ADP + (phosphate)n+1
-
 r
ATP + (phosphate)n PPK2B forms polyphopshate with an average chain length of about 125 Corynebacterium glutamicum MH20-22B ADP + (phosphate)n+1
-
 r
ATP + (phosphate)n PPK2B forms polyphopshate with an average chain length of about 125 and prefers the polyphosphate synthesis reaction direction Corynebacterium glutamicum MH20-22B ADP + (phosphate)n+1 polyphosphate product chain length determination by NMR spectroscopy r
ATP + ATP polyP not required as primer Corynebacterium glutamicum ADP + (phosphate)n+1
-
 r
GTP + (phosphate)n
-
 Corynebacterium glutamicum GDP + (phosphate)n+1
-
 r
GTP + (phosphate)n lower activity compared to ATP Corynebacterium glutamicum GDP + (phosphate)n+1
-
 r
ITP + (phosphate)n
-
 Corynebacterium glutamicum IDP + (phosphate)n+1
-
 r
ITP + (phosphate)n lower activity compared to ATP Corynebacterium glutamicum IDP + (phosphate)n+1
-
 r
additional information accumulation of high levels of cytosolic phosphorus in the form of polyphosphate involving classII polyphosphate kinase isozymes, granular poly P, i.e. volutin, can make up to 37% of the internal cell volume, overview Corynebacterium glutamicum ?
-
 ?
additional information PKK2B also shows NDP kinase activity Corynebacterium glutamicum ?
-
 ?
additional information accumulation of high levels of cytosolic phosphorus in the form of polyphosphate involving classII polyphosphate kinase isozymes, granular poly P, i.e. volutin, can make up to 37% of the internal cell volume, overview Corynebacterium glutamicum MH20-22B ?
-
 ?
additional information PKK2B also shows NDP kinase activity Corynebacterium glutamicum MH20-22B ?
-
 ?

Subunits

Subunits Comment Organism
homotetramer 4 x 39000, SDS PAGE Corynebacterium glutamicum
tetramer PPK2B is active as a homotetramer Corynebacterium glutamicum

Synonyms

Synonyms Comment Organism
ATP:polyphosphate phosphotransferase
-
 Corynebacterium glutamicum
class II polyphosphate kinase
-
 Corynebacterium glutamicum
kinase, polyphosphate (phosphorylating)
-
 Corynebacterium glutamicum
polyphosphate kinase
-
 Corynebacterium glutamicum
PPK
-
 Corynebacterium glutamicum
PPK2
-
 Corynebacterium glutamicum
PPK2B
-
 Corynebacterium glutamicum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Corynebacterium glutamicum

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
20
-
 preincubation results in a loss of 50% of the activity Corynebacterium glutamicum
20 25 30 min, purified recombinant wild-type PKK2B, 50% remaining activity Corynebacterium glutamicum
25
-
 preincubation results in a loss of 50% of the activity Corynebacterium glutamicum
45
-
 preincubation results in a loss of 50% of the activity with substrate Corynebacterium glutamicum
45
-
 preincubation results in an almost complete loss the activity without substrate Corynebacterium glutamicum
45
-
 30 min, purified recombinant wild-type PKK2B, almost no remaining activity in absence of polyphosphate, but about 50% remaining activity in presence of 40 mM polyphosphate Corynebacterium glutamicum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
34
-
 GTP pH 7.2, 30°C, recombinant wild-type enzyme Corynebacterium glutamicum
43
-
 ITP pH 7.2, 30°C, recombinant wild-type enzyme Corynebacterium glutamicum
74
-
 ATP pH 7.2, 30°C, recombinant wild-type enzyme Corynebacterium glutamicum
77
-
 Polyphosphate pH 7.2, 30°C, recombinant wild-type enzyme Corynebacterium glutamicum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.2
-
 narrow optimum Corynebacterium glutamicum
7.2
-
 in 50mM TEA Corynebacterium glutamicum

Cofactor

Cofactor Comment Organism Structure
ATP
-
 Corynebacterium glutamicum

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.9
-
 GMP
-
 Corynebacterium glutamicum
3.9
-
 GMP pH 7.2, 30°C, recombinant wild-type enzyme Corynebacterium glutamicum
6.6
-
 IMP
-
 Corynebacterium glutamicum
6.6
-
 IMP pH 7.2, 30°C, recombinant wild-type enzyme Corynebacterium glutamicum
7.9
-
 AMP no inhibition by CMP and UMP Corynebacterium glutamicum
7.9
-
 AMP pH 7.2, 30°C, recombinant wild-type enzyme Corynebacterium glutamicum