Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 3.1.3.32 extracted from

  • Keppetipola, N.; Shuman, S.
    Characterization of the 2,3 cyclic phosphodiesterase activities of Clostridium thermocellum polynucleotide kinase-phosphatase and bacteriophage lambda phosphatase (2007), Nucleic Acids Res., 35, 7721-7732.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.1.3.32

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Acetivibrio thermocellus

Protein Variants

Protein Variants Comment Organism
D233E decreased activity compared to the wild type enzyme for hydrolysis of 2',3'-cAMP Acetivibrio thermocellus
D236A 64% as active as wild-type Pnkp in cleaving 2',3'-cAMP Acetivibrio thermocellus
D236E decreased activity compared to the wild type enzyme for hydrolysis of 2',3'-cAMP Acetivibrio thermocellus
D236N decreased activity compared to the wild type enzyme for hydrolysis of 2',3'-cAMP Acetivibrio thermocellus
D392A 7-10% as active as wild type Pnkp Acetivibrio thermocellus
D392E increased activity compared to the wild type enzyme for hydrolysis of 2',3'-cAMP Acetivibrio thermocellus
D392N increased activity compared to the wild type enzyme for hydrolysis of 2',3'-cAMP Acetivibrio thermocellus
H189A 17% as active as wild type Pnkp Acetivibrio thermocellus
H189A decreased activity compared to the wild type enzyme for hydrolysis of 2',3'-cAMP Acetivibrio thermocellus
H189D the mutation imposes strict specificity for a 2',3' cyclic phosphate, which is cleaved to form a single 2'-NMP product and shows increased activity compared to the wild type enzyme Acetivibrio thermocellus
H189E increased activity compared to the wild type enzyme for hydrolysis of 2',3'-cAMP Acetivibrio thermocellus
H264A 10% as active as wild-type Pnkp in cleaving 2',3'-cAMP Acetivibrio thermocellus
H323A 60% of wild type activity Acetivibrio thermocellus
H323Q decreased activity compared to the wild type enzyme for hydrolysis of 2',3'-cAMP Acetivibrio thermocellus
H376A 7-10% as active as wild type Pnkp Acetivibrio thermocellus
H376D decreased activity compared to the wild type enzyme for hydrolysis of 2',3'-cAMP Acetivibrio thermocellus
H376N decreased activity compared to the wild type enzyme for hydrolysis of 2',3'-cAMP Acetivibrio thermocellus
N263A 7-10% as active as wild type Pnkp Acetivibrio thermocellus
R237A 24% as active as wild-type Pnkp in cleaving 2',3'-cAMP Acetivibrio thermocellus
R237K increased activity compared to the wild type enzyme for hydrolysis of 2',3'-cAMP Acetivibrio thermocellus
R237Q increased activity compared to the wild type enzyme for hydrolysis of 2',3'-cAMP Acetivibrio thermocellus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.9
-
 2',3'-cAMP mutant enzyme H376D, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
4.7
-
 2',3'-cAMP mutant enzyme H376N, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
6.6
-
 2',3'-cAMP mutant enzyme D236N, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
7.8
-
 2',3'-cAMP mutant enzyme D236A, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
18
-
 2',3'-cAMP mutant enzyme D236E, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
18
-
 2',3'-cAMP mutant enzyme D392E, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
18
-
 2',3'-cAMP wild type enzyme, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
20
-
 2',3'-cAMP mutant enzyme H189D, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
24
-
 2',3'-cAMP mutant enzyme H323Q, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
28
-
 2',3'-cAMP mutant enzyme H189E, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
29
-
 2',3'-cAMP mutant enzyme H189A, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
31
-
 2',3'-cAMP mutant enzyme D392N, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
32
-
 2',3'-cAMP mutant enzyme H323A, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
60
-
 2',3'-cAMP mutant enzyme R237Q, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
62
-
 2',3'-cAMP mutant enzyme D233E, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
67
-
 p-Nitrophenyl phenylphosphonate wild type enzyme, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
77
-
 2',3'-cAMP mutant enzyme R237A, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
100
-
 2',3'-cAMP mutant enzyme D392A, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
100
-
 2',3'-cAMP mutant enzyme R237K, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ 0.5mM, supports one-third the activity of Mn2+ Acetivibrio thermocellus
Mn2+ 0.5 mM, dependent Acetivibrio thermocellus
additional information Mg2+, Ca2+, Co2+, Cd2+, Co2+, and Zn2+ have no significant influence on activity Acetivibrio thermocellus
Ni2+ 0.5mM, supports one-third the activity of Mn2+ Acetivibrio thermocellus

Organism

Organism UniProt Comment Textmining
Acetivibrio thermocellus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-agarose chromatography Acetivibrio thermocellus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2',3'-cAMP + H2O
-
 Acetivibrio thermocellus ?
-
 ?
bis-p-nitrophenyl phosphate + H2O
-
 Acetivibrio thermocellus ?
-
 ?
additional information not active towards dimethyl-p-nitrophenyl phosphate, thymidine 5'-monophosphate-p-nitrophenyl ester, and p-nitrophenyl phosphorylcholine Acetivibrio thermocellus ?
-
 ?
p-nitrophenyl phenylphosphonate + H2O
-
 Acetivibrio thermocellus ?
-
 ?

Synonyms

Synonyms Comment Organism
PNKP
-
 Acetivibrio thermocellus
polynucleotide kinase-phosphatase prefers a 2',3' cyclic phosphate to a 3',5'cyclic phosphate Acetivibrio thermocellus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
additional information
-
 2',3'-cAMP mutant enzyme D236E, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
0.04 1.97 2',3'-cAMP mutant enzyme R237A, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
0.04 1.97 2',3'-cAMP wild type enzyme, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
0.833
-
 2',3'-cAMP mutant enzyme D392E, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
0.97
-
 2',3'-cAMP mutant enzyme H189E, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
1.1
-
 2',3'-cAMP mutant enzyme D392N, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
1.45
-
 2',3'-cAMP mutant enzyme H189A, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
1.67
-
 2',3'-cAMP mutant enzyme D392A, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
3.2
-
 2',3'-cAMP mutant enzyme H376D, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
3.25
-
 2',3'-cAMP mutant enzyme D233E, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
3.5
-
 2',3'-cAMP mutant enzyme R237Q, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
4.53
-
 2',3'-cAMP mutant enzyme D236A, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
5.13
-
 2',3'-cAMP mutant enzyme H376N, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
5.77
-
 2',3'-cAMP mutant enzyme D236N, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
6.03
-
 2',3'-cAMP mutant enzyme D236E, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
6.5
-
 2',3'-cAMP mutant enzyme H323Q, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
8.1
-
 2',3'-cAMP mutant enzyme R237K, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
8.55
-
 2',3'-cAMP mutant enzyme R237A, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
8.93
-
 2',3'-cAMP wild type enzyme, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
9.1
-
 2',3'-cAMP mutant enzyme H323A, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
15.97
-
 2',3'-cAMP mutant enzyme D392E, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
17.17
-
 2',3'-cAMP mutant enzyme H189E, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
18.67
-
 2',3'-cAMP mutant enzyme D392N, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
20.17
-
 2',3'-cAMP mutant enzyme R237Q, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
26.17
-
 2',3'-cAMP mutant enzyme R237K, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
27.83
-
 p-Nitrophenyl phenylphosphonate wild type enzyme, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus
34.33
-
 2',3'-cAMP mutant enzyme H189D, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2, at 45°C Acetivibrio thermocellus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
 in the presence of Mn2+ Acetivibrio thermocellus

pH Stability

pH Stability pH Stability Maximum Comment Organism
5.5 8 activity declines sharply at less than pH 5.5 or above pH 8.0 Acetivibrio thermocellus