Information on EC 3.6.1.11 - exopolyphosphatase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea

EC NUMBER
COMMENTARY hide
3.6.1.11
-
RECOMMENDED NAME
GeneOntology No.
exopolyphosphatase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(polyphosphate)n + H2O = (polyphosphate)n-1 + phosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of phosphoric ester
-
phosphorous acid anhydride hydrolysis
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
non-pathway related
-
-
Purine metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
polyphosphate phosphohydrolase
-
CAS REGISTRY NUMBER
COMMENTARY hide
9024-85-5
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
promastigotes
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
NM-1 strain, two isoenzymes polyphosphatases I and II
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
strain 8830
SwissProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain AH22
-
-
Manually annotated by BRENDA team
strain CRX
-
-
Manually annotated by BRENDA team
strain CRY
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(phosphate)13-18 + H2O
(phosphate)12-17 + phosphate
show the reaction diagram
(phosphate)15 + H2O
(phosphate)14 + phosphate
show the reaction diagram
(phosphate)208 + H2O
(phosphate)207 + phosphate
show the reaction diagram
(phosphate)25 + H2O
(phosphate)24 + phosphate
show the reaction diagram
(phosphate)3 + H2O
(phosphate)2 + phosphate
show the reaction diagram
(phosphate)300 + H2O
(phosphate)299 + phosphate
show the reaction diagram
(phosphate)4 + H2O
(phosphate)3 + phosphate
show the reaction diagram
-
-
-
-
?
(phosphate)45 + H2O
(phosphate)44 + phosphate
show the reaction diagram
(phosphate)65 + H2O
(phosphate)64 + phosphate
show the reaction diagram
(phosphate)9 + H2O
(phosphate)8 + phosphate
show the reaction diagram
45% of the activity with (phosphate)25
-
-
?
(phosphate)n + H2O
(phosphate)n-1 + phosphate
show the reaction diagram
(polyphosphate)130 + H2O
(polyphosphate)129 + phosphate
show the reaction diagram
(polyphosphate)14 + H2O
(polyphosphate)13 + phosphate
show the reaction diagram
(polyphosphate)60 + H2O
(polyphosphate)59 + phosphate
show the reaction diagram
(polyphosphate)n + H2O
(polyphosphate)n-1 + phosphate
show the reaction diagram
3'-AMP + H2O
adenosine + phosphate
show the reaction diagram
-
-
-
-
3'-CMP + H2O
cytosine + phosphate
show the reaction diagram
-
-
-
-
5'-AMP + H2O
adenosine + phosphate
show the reaction diagram
-
-
-
-
5'-dGMP + H2O
deoxyguanosine + phosphate
show the reaction diagram
-
-
-
-
5'-GMP + H2O
guanosine + phosphate
show the reaction diagram
-
-
-
-
adenosine 5'-pentaphosphate + H2O
?
show the reaction diagram
-
-
-
-
?
adenosine 5'-tetraphosphate + H2O
ATP + phosphate
show the reaction diagram
guanosine 5'-tetraphosphate + H2O
?
show the reaction diagram
guanosine 5'-tetraphosphate + H2O
GTP + phosphate
show the reaction diagram
guanosine-5'-tetraphosphate + H2O
GTP + phosphate
show the reaction diagram
inosine tetraphosphate + H2O
ITP + phosphate
show the reaction diagram
-
-
-
?
p-nitrophenyl phosphate + H2O
p-nitrophenol + phosphate
show the reaction diagram
-
-
-
-
pentasodium triphosphate + H2O
pentasodium diphosphate + phosphate
show the reaction diagram
polyphosphate + H2O
?
show the reaction diagram
-
chain length of more than 45 phosphate residues
-
-
?
polyphosphate 15 + H2O
polyphosphate 14 + phosphate
show the reaction diagram
polyphosphate 188 + H2O
polyphosphate 187 + phosphate
show the reaction diagram
-
-
-
-
?
polyphosphate 208 + H2O
polyphosphate 207 + phosphate
show the reaction diagram
polyphosphate 3 + H2O
polyphosphate 2 + phosphate
show the reaction diagram
polyphosphate 75 + H2O
polyphosphate 74 + phosphate
show the reaction diagram
-
-
-
-
?
sodium phosphate glass type 15 + H2O
? + phosphate
show the reaction diagram
-
-
-
-
?
tetraphosphate + H2O
triphosphate + phosphate
show the reaction diagram
-
-
-
?
triphosphate + H2O
?
show the reaction diagram
-
-
-
?
triphosphate + H2O
diphosphate + phosphate
show the reaction diagram
tripolyphosphate + H2O
phosphate + ?
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(phosphate)n + H2O
(phosphate)n-1 + phosphate
show the reaction diagram
(polyphosphate)n + H2O
(polyphosphate)n-1 + phosphate
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
in the presence of Ca2+, the activity is about 20% of levels with Mg2+
Cd2+
-
stimulated in the micromolar range to a lower extent than Cu2+ and Mn2+
Cu2+
-
0.01 mM, stimulates
NaCl
-
50 and 200 mM, 46 and 42% activity enhancement, respectively
NH4+
100 mM, 35% activity enhancement; slight activation
NH4Cl
-
50 and 200 mM, 42 and 67% activity enhancement, respectively
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(phosphate)25
-
-
-
(phosphate)45
-
-
-
(phosphate)65
-
-
-
AlF4-
-
10 mM, 25% loss of activity
Ammonium molybdate
-
-
arginine
inhibition of the recombinant enzyme at high concentrations
CaCl2
3.8 mM CaCl2 leads to half-maximal inhibition of His-tagged PPX2
CHES buffer
-
20 mM
diphosphate
dipyridamole
-
known inhibitor of phosphodiesterase, no effect on the triphosphate hydrolyzing activity of h-prune
dithiothreitol
-
0.1 mM, 13% activity loss
Guanosine 5'-pentaphosphate
1 mM, 73% resiudal activity
guanosine 5'-tetraphosphate
1 mM, 68% resiudal activity
heparin
hydrogen peroxide
-
-
iodoacetamide
KCl
slight inhibition at high concentrations
long-chain polyphosphate
-
potential physiological regulator, inhibits h-prune-catalyzed hydrolysis of triphosphate
-
lysine
inhibition of the recombinant enzyme at high concentrations
MES buffer
-
20 mM
-
Mn2+
PPX2 is inhibited by millimolar concentrations, activity is reduced 2fold at 3.5 mM MnCl2
nm23-H1
-
metastasis suppressor protein, the exopolyphosphatase activity is suppressed
-
o-vanadate
phosphate
Polyphosphate
-
polyP15, 50% inhibition at 200fold polymer molar excess over the long-chain polyphosphate substrate
polyphosphate 3
-
1mM, 4% activity loss
-
potassium cyanide
-
-
Pyrophosphate
8.2 mM pyrophosphate leads to half-maximal inhibition of His-tagged PPX2
spermidine
-
0.17 mM, 50% loss of activity
Tetrapolyphosphate
-
50% inhibition at 2000fold polymer molar excess over the long-chain polyP substrate
tripolyphosphate
VO43-
-
1 mM, less than 5% reduced activity
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(NH4)2SO4
-
50 mM, stimulates
ADP
-
slight stimulation
ammonium chloride
200 mM, stimulates almost twofold in the presence and absence of 0.1 mM Co2+
arginine
-
stimulates
carbonyl cyanide-4-trifluoromethoxyphenylhydrazone
-
-
dithiothreitol
-
about 50% stimulation of activity at 1 mM
KCl
-
150-300 mM
NADH
-
2fold activation
polylysine
100 mg/l, 2fold activity enhancement
Pyruvic acid
-
activates PPX1, activation is inhibited KCN
succinic acid
-
activates PPX1, activation is inhibited KCN
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.264 - 0.597
(phosphate)13-18
0.0011 - 0.0028
(phosphate)15
0.0069
(phosphate)25
-
2 mM Mg2+ as cofactor, 0.1 M Tris-HCl, pH 7.2, 0.05 mM EGTA
-
0.0002 - 0.213
(phosphate)3
0.019 - 0.041
(phosphate)4
0.0022
(phosphate)45
-
2 mM Mg2+ as cofactor, 0.1 M Tris-HCl, pH 7.2, 0.05 mM EGTA
-
0.0007 - 0.0036
(phosphate)65
0.0173
(polyphosphate)130
pH 6.8, 37C
-
0.0059
(polyphosphate)14
pH 6.8, 37C
-
0.0121
(polyphosphate)60
pH 6.8, 37C
-
0.0035 - 1343
(Polyphosphate)n
0.1
3'-AMP
+/- 0.01
0.37
3'-CMP
+/- 0.08
0.32
5'-AMP
+/- 0.04
0.28
5'-dGMP
+/- 0.04
0.26
5'-GMP
+/- 0.05
0.028 - 0.08
adenosine 5'-tetraphosphate
0.1
adenosine-5'-tetraphosphate
-
-
0.012 - 54.5
guanosine 5'-tetraphosphate
0.08
guanosine-5'-tetraphosphate
-
-
2.49
p-nitrophenyl phosphate
+/- 0.4
0.0272
pentasodium triphosphate
in 50 mM HEPES, pH 7.8, 0.05 mM EGTA,1 mM MgCl2, at 30C
0.0039 - 0.0238
polyP10
0.000024
polyP100
-
-
0.011 - 0.093
polyP15
0.0012 - 0.0024
polyP208
0.00016
polyP25
-
-
0.000004
polyP250
-
-
0.0013
polyP33-36
-
-
-
0.00006
polyP50
-
-
0.00005
polyP500
-
-
0.033
polyP9-10
-
-
-
0.0035 - 12
Polyphosphate
75
polyphosphate 15
-
-
-
3.5
polyphosphate 208
-
-
-
1100
polyphosphate 3
-
-
-
0.0022
sodium phosphate glass type 15
-
in 50 mM Tris-HCl buffer (pH 7.2), at 28C
-
0.11
Tetraphosphate
in 50 mM PIPES, pH 6.8 containing 25 mM KCl and 2 mM MgCl2
0.0002 - 0.058
Triphosphate
0.14 - 0.41
tripolyphosphate
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
157 - 458
(phosphate)13-18
0.16
(phosphate)25
Homo sapiens
-
2 mM Mg2+ as cofactor, 0.1 M Tris-HCl, pH 7.2, 0.05 mM EGTA
-
0.57 - 399
(phosphate)3
3.4 - 7.1
(phosphate)4
0.22
(phosphate)45
Homo sapiens
-
2 mM Mg2+ as cofactor, 0.1 M Tris-HCl, pH 7.2, 0.05 mM EGTA
-
0.03
(phosphate)65
Homo sapiens
-
2 mM Mg2+ as cofactor, 0.1 M Tris-HCl, pH 7.2, 0.05 mM EGTA
-
1550
(Phosphate)n
Saccharomyces cerevisiae
-
37C, recombinant enzyme
6.7
(polyphosphate)130
Mycobacterium tuberculosis
P9WHV5
pH 6.8, 37C
-
10.2
(polyphosphate)14
Mycobacterium tuberculosis
P9WHV5
pH 6.8, 37C
-
7.3
(polyphosphate)60
Mycobacterium tuberculosis
P9WHV5
pH 6.8, 37C
-
9.84
3'-AMP
Escherichia coli
P0A840
+/- 0.37
5.93
3'-CMP
Escherichia coli
P0A840
+/- 0.44
4.9
5'-AMP
Escherichia coli
P0A840
+/- 0.3
8.04
5'-dGMP
Escherichia coli
P0A840
+/- 0.39
11
5'-GMP
Escherichia coli
P0A840
+/- 0.87
31 - 723
adenosine 5'-tetraphosphate
40
adenosine-5'-pentaphosphate
Saccharomyces cerevisiae
-
-
0.7 - 308
guanosine 5'-tetraphosphate
3.55
p-nitrophenyl phosphate
Escherichia coli
P0A840
+/- 0.16
8.1
pentasodium triphosphate
Trypanosoma brucei
Q7Z032
in 50 mM HEPES, pH 7.8, 0.05 mM EGTA,1 mM MgCl2, at 30C
63
polyP10
Saccharomyces cerevisiae
-
-
3.3
polyP100
Saccharomyces cerevisiae
-
-
15
polyP25
Saccharomyces cerevisiae
-
-
1.2
polyP250
Saccharomyces cerevisiae
-
-
81
polyP33-36
Saccharomyces cerevisiae
-
-
-
8.4
polyP50
Saccharomyces cerevisiae
-
-
1
polyP500
Saccharomyces cerevisiae
-
-
90
polyP9-10
Saccharomyces cerevisiae
-
-
-
0.05 - 1150
Polyphosphate
0.9
Tetraphosphate
Corynebacterium glutamicum
Q8NRR8, Q8NT99
in 50 mM PIPES, pH 6.8 containing 25 mM KCl and 2 mM MgCl2
0.6 - 3.46
Triphosphate
143 - 180
tripolyphosphate
additional information
additional information
Escherichia coli
-
-
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
263 - 1716
(phosphate)13-18
63 - 4081
(phosphate)3
387
(polyphosphate)130
Mycobacterium tuberculosis
P9WHV5
pH 6.8, 37C
206948
1729
(polyphosphate)14
Mycobacterium tuberculosis
P9WHV5
pH 6.8, 37C
206946
603
(polyphosphate)60
Mycobacterium tuberculosis
P9WHV5
pH 6.8, 37C
206947
42 - 1273
guanosine 5'-tetraphosphate
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.6 - 24.1
ATP
0.8
GTP
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.163 - 0.33
(phosphate)25
0.055 - 0.12
(phosphate)45
0.064 - 0.105
(phosphate)65
1.2 - 5.7
diphosphate
1
hydrogen peroxide
Rhipicephalus microplus
-
in 50 mM Tris-HCl buffer (pH 7.2), at 28C
0.0019 - 0.0072
nm23-H1
0.0213
Zn2+
Trypanosoma brucei
Q7Z032
in 50 mM HEPES, pH 7.8, 0.05 mM EGTA,1 mM MgCl2, at 30C
0.032
ZnCl2
Homo sapiens
-
2 mM Mg2+, hydrolysis of triphospate is inhibited
additional information
heparin
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.006
-
strain CRX with inactivated ppx1 and ppN1 gene, cytosol preparation in exponential growth phase in phosphate-deficient medium
0.01
-
strain CRX with inactivated ppx1 and ppN1 gene, cytosol preparation in exponential growth phase in phosphate-rich medium
0.014
-
40-kDa-exopolyphosphatase from Saccharomyces cerevisiae strain CRN grown on glucose
0.034
-
membrane-bound form of exopolyphospatase from Saccharomyces cerevisiae strain CRX grown on lactate; membrane-bound form of exopolyphospatase from Saccharomyces cerevisiae strain CRY grown on lactate
0.035
-
high-molecular weight exopolyphosphatase from Saccharomyces cerevisiae strain CRX grown on glucose
0.05
1.25 mM polyphosphate 3 as substrate, in the presence of 2.5 mM Mg2+
0.055
-
strain CRX with inactivated ppx1 gene, cytosol preparation in exponential growth phase in phosphate-deficient medium
0.058
-
soluble form of exopolyphospatase from Saccharomyces cerevisiae strain CRX grown on lactate
0.07
-
strain CRX with inactivated ppn1 gene, cytosol preparation in exponential growth phase in phosphate-rich medium
0.097
-
membrane-bound exopolyphosphatase from Saccharomyces cerevisiae strain CRY grown on glucose
0.119
-
soluble form of exopolyphospatase from Saccharomyces cerevisiae strain CRY grown on lactate
0.12
wild type, samples containing 50 mM PIPES, pH 6.8, 25 mM KCl, and 2 mM MgCl2; wild type, samples containing 50 mM PIPES, pH 6.8, 25 mM KCl, and 2 mM MgCl2
0.136
-
40-kDa-exopolyphosphatase from Saccharomyces cerevisiae strain CRY grown on glucose
0.18
-
parent strain CRY, cytosol preparation in exponential growth phase in phosphate-rich medium
0.21
0.13 mM polyphosphate 15 as substrate, in the presence of 1 mM Co2+
0.23
0.13 mM polyphosphate 15 as substrate, in the presence of 0.1 mM Co2+
0.28
0.01 mM polyphosphate 208 as substrate, in the presence of 1 mM Co2+
0.3
2.0 mM polyphosphate 208 as substrate, in the presence of 0.1 mM Co2+
0.43
overexpression of ppx1 increase the specific activity of exopolyphosphatase by 4fold compared to that of the empty vector control
0.47
-
enzyme from cystosol
0.8
crude cell extracts of wild type strain pVWEx1-ppx2 show 6fold higher exopolyphosphatase activity than those of wild type pVWEx1
1.11
-
after ammonium sulfate precipitation
1.26
-
mitochondrial fraction
3 - 8
substrate (phosphate)3, presence of 0.1 mM Co2+, pH 7.2, 30C
5
-
after DEAE-Toyopearl 650 M column chromatography
7.24
+/- 0.34, p-nitrophenyl phosphate
10
+/- 0.61, 5'-AMP
12.1
+/- 0.90, 3'-CMP
15
substrate (phosphate)300, pH 6.5, 25C
16.4
+/- 0.80, 5'-dGMP
20.1
+/- 0.76, 3'-AMP
22.4
+/- 1.78, 5'-GMP
126
substrate (phosphate)300, pH 6.5, 25C
150
-
after 319fold purification with heparin agarose, at 30C in 1 ml of reaction mixture containing 50 mM Tris-HCl buffer, pH 7.2, 0.1 mM CoSO4, 200 mM ammonium chloride, and 0.01 mM polyphosphate 208; pH 7.2, 30C, polyP208 as substrate
170
substrate (phosphate)13-18, pH 6.5, 25C
180
substrate (phosphate)13-18, pH 6.5, 25C
240
substrate (phosphate)15, presence of 0.1 mM Co2+, pH 7.2, 30C
290
substrate (phosphate)208, presence of 0.1 mM Co2+, pH 7.2, 30C
590
substrate (phosphate)3, pH 6.5, 25C
2070
-
purified recombinant enzyme
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5
-
polyphosphatase I
4.8
-
in presence of Co2+
5
-
exopolyphosphatase I
6 - 6.5
-
and a second higher optimum at pH 7.5
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.8 - 7.2
-
50% of maximal activity at pH 3.8 and at pH 7.2
4.6 - 5.4
-
pH 4.6: 81% of maximal activity, pH 5.4: 59% of maximal activity, exopolyphosphatase I
5.5 - 7.5
more than 30-40% of maximum activity
6 - 9
-
45-55% of maximal activity at pH 6.0 and at pH 9.0
6.2 - 8.4
-
pH 6.2: about 35% of maximal activity, pH 8.4: about 60% of maximal activity
7 - 9
-
pH 7.0: 5% of maximal activity, pH 9.0: 57% of maximal activity, exopolyphosphatase I
10
about 90% of maximum activity
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 48
-
30C: about about 60% of maximal activity, 48C: about 40% of maximal activity
30 - 53
-
30C: about 30% of maximal activity, 53C: about 40% of maximal activity
70
75% of maximum activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.4
calculated from amino acid sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
activity is 134% greater in mycorrhizal roots than in non-mycorrhizal roots of Allium cepa
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
additional information
PDB
SCOP
CATH
ORGANISM
UNIPROT
Aquifex aeolicus (strain VF5)
Aquifex aeolicus (strain VF5)
Cytophaga hutchinsonii (strain ATCC 33406 / NCIMB 9469)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
28000
-
gel filtration
33000
-
SDS-PAGE, probably a product of mRNA splicing or/and protein processing
35000
-
gel filtration
38800
gel filtration
42870
calculated from amino acid sequence
50000
-
estimated from the amino acid composition
55000
-
polyphosphatase II, gel filtration
58100
-
SDS-PAGE
70000
-
exopolyphosphatase II, gel filtration
78000
-
MALDI-TOF
93000
-
polyphosphatase I, gel filtration
100000
-
gel filtration
100400
gel filtration
120000 - 830000
-
PPN1
125000
-
Superose 6 column gel filtration, active but unstable enzyme complex
200000
gel filtration; gel filtration on Superose 6
245000
-
gel filtration
450000
-
determined by gel filtration
500000
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
oligomer
at least four subunits in solution
tetramer
-
4 * 55000, SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
PPN1 gene encodes a polypeptide with the predicted molecular mass of 78 kDa, the monomer molecular mass of the mature PPN1 is about 33-35 kDa
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop vapour diffusion method
-
sitting drop vapour diffusion method
-
enzxme in complex with phosphate, sulfate, or ATP, X-ray diffraction structure determination and analysis at 1.6, 1.8, and 1.9 A resolution, multiple isomorphous replacement with anomalous scattering technique
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
-
4C, 48 h, 95% loss of activity
209846
5
-
4C, 48 h, 66% loss of activity
209846
6
-
4C, 48 h, 21% loss of activity
209846
7
-
4C, 48 h, 5% loss of activity
209846
7.5 - 8
-
4C, 48 h, stable
209846
9
-
4C, 48 h, 11% loss of activity
209846
10
-
4C, 48 h, 95% loss of activity
209846
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0
-
incubation of h-prune 1-100 microM for 5 h in the presence of 0.1 M Tris-HCl, pH 7.2, and 0.05 mM EGTA inactivates the enzyme 2-4fold, no inactivation is evident in the presence of 1 mM Mg2+
25 - 40
at pH 6.8, 2 mM MgCl2 and 25 mM KCl, no significant loss of activity is observed after 60 min
40
-
15 min, 70% loss of activity
45
after preheating at 45C for 30 and 60 min, 70% and 40% of the PPX2 activity remains
50
after preheating at 50C, His-tagged PPX2 loses its activity quickly, after incubation for 60 min, no activity remains
70
-
10 min, inactivation
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
addition of 0.05 mM CoSO4 does not increase the enzyme stability independent of the presence of Triton X-100
-
loss of activity after dialysis
-
Mg2+ stabilizes h-prune against inactivation during storage
-
more than 90% loss of activity after freezing in liquid nitrogen and subsequent thawing
-
repeated freezing and thawing destroys activity
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 50 mM Tris-HCl buffer, pH 7.6, 10 mM MgCl2, 0.5 mM EDTA, 150 mM NaCl, 20% loss of activity after 2 months
-
-20C, in 50 mM Tris-HCl buffer, pH 7.5, 5 mM MgCl2, 0.5 mM EDTA, 50 mM NaCl, 0.1% w/v bovine serum albumin, 40% v/v glycerol, exopolyphosphatase I and II, less than 30% loss of activity after 2 months
-
-20C, stable for months
-
-4C, 0.1% Triton X-100, 1M KCl, 4 days, no activity loss
-
-4C, enzyme desorbed from heparin-agarose with 1 M KCl and 0.1% Triton X-100, 4 days, no loss of activity
-
-80C, 300 mM NaCl, 250 mM sucrose, 2 mM dithiothreitiol, several months, no activity loss
-
4C, 5% glycerol, 0.5 M NaCl, pH 7.5, no loss in activity after several months
4C, enzyme obtained by desorption with polyphosphate, 2 h, 30% loss of activity
-
4C, in absence of detergents, complete loss of activity after 24 h, can be stabilized by 0.1% Triton X-100 and protease inhibitors
-
92C, in the presence of 0.1% Triton X-100, 5 days, 8% loss of activity of the partially purified exopolyphosphatase preparation after DEAE-Toyopearl chromatography
-
92C, without 0.1% Triton X-100, 5 days, 66% loss of activity of the partially purified exopolyphosphatase preparation after DEAE-Toyopearl chromatography
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by metal chelate chromatography
-
DEAE Sepharose filtration
-
DEAE-Toyopearl 650 M column chromatography; partial, using ion-exchange chromatography and gel filtration
-
exopolyphosphatase I and II
-
gel filtration
-
glutathione-Sepharose resin column chromatography or amylose-resin column chromatography
HP-SP-Sepharose column chromatography
-
ion-exchange chromatography
isolation of a recombinant His-tagged protein by affinity chromatography followed by cleavage of the polyhistidine tag
-
isolation of two isoenzymes by cellular fractionation and gel filtration
-
recombinant PPX1 17.4fold in a two-step procedure
-
recombinant ppx2 enzyme is purified by Ni-nitrilotriacetic acid chromatography
recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3)
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, functional expression in Escherichia coli strain BL21(DE3)
expressed in Escherichia coli Arctic Express (DE3) cells
expressed in Escherichia coli as His-tagged wild type protein and His-tagged variants
-
expressed in Escherichia coli BL21(DE3) cells
expression in Escherichia coli
expression in Escherichia coli of a his-tagged recombinant enzyme
-
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; 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
expression in Escherichia coli; expression in Escherichia coli
expression in Saccharomyces cerevisiae
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
-
gene PPX1, overexpression
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
under nitrogen-limiting conditions, exopolyphosphatase expression is controlled from a sigma54-dependent promoter activated by the response regulator NtrC. Under phosphate limitation, the expression is controlled from a sigma70 promoter, activated by PhoB
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D143A
-
single amino acid mutation of Ppx
E121A
-
single amino acid mutation of Ppx
E150A
-
single amino acid mutation of Ppx
E371A
-
single amino acid mutation of Ppx
D106A
-
variant displays reduced activity with a turnover value of 35% compared to the wild type counterpart
D179A
-
variant is inactive
D28A
-
variant is inactive
H107N
-
variant displays reduced activity with a turnover value of 4.4% compared to the wild type counterpart, Km value increases 7fold
H108N
-
variant displays reduced activity with a turnover value of 32% compared to the wild type counterpart
N24H
-
variant is inactive
R128H
-
enhanced kcat value (146%) is obtained with the mutant protein compared to the wild type counterpart, Km value increases 21fold
R348A
-
variant is inactive
D127E
-
site-directed mutagenesis, the mutant shows reduced the Mg2+ affinity of the tight binding site compared to the wild-type enzyme, the activaion by divalent cations differs between wild-type and mutant enzymes, overview
D127N
-
site-directed mutagenesis, the mutant shows reduced the Mg2+ affinity of the tight binding site compared to the wild-type enzyme, the activaion by divalent cations differs between wild-type and mutant enzymes, overview
H148N
-
site-directed mutagenesis, the mutant shows increased Km and reduced kcat in comparison to the wild-type enzyme, the activaion by divalent cations differs between wild-type and mutant enzymes, overview
H149N
-
site-directed mutagenesis, the mutant shows increased Km and reduced kcat in comparison to the wild-type enzyme, the activaion by divalent cations differs between wild-type and mutant enzymes, overview
N35H
-
site-directed mutagenesis, the mutant shows reduced the Mg2+ affinity of the tight binding site compared to the wild-type enzyme, the activation by divalent cations differs between wild-type and mutant enzymes, overview
D127E
-
site-directed mutagenesis, the mutant shows reduced the Mg2+ affinity of the tight binding site compared to the wild-type enzyme, the activaion by divalent cations differs between wild-type and mutant enzymes, overview
-
D127N
-
site-directed mutagenesis, the mutant shows reduced the Mg2+ affinity of the tight binding site compared to the wild-type enzyme, the activaion by divalent cations differs between wild-type and mutant enzymes, overview
-
H148N
-
site-directed mutagenesis, the mutant shows increased Km and reduced kcat in comparison to the wild-type enzyme, the activaion by divalent cations differs between wild-type and mutant enzymes, overview
-
N35H
-
site-directed mutagenesis, the mutant shows reduced the Mg2+ affinity of the tight binding site compared to the wild-type enzyme, the activation by divalent cations differs between wild-type and mutant enzymes, overview
-
additional information