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Literature summary for 3.1.13.3 extracted from

  • Young Park, A.; Elvin, C.M.; Hamdan, S.M.; Wood, R.J.; Liyou, N.E.; Hamwood, T.E.; Jennings, P.A.; Dixon, N.E.
    Hydrolysis of the 5-p-nitrophenyl ester of TMP by oligoribonucleases (ORN) from Escherichia coli, Mycobacterium smegmatis, and human (2008), Protein Expr. Purif., 57, 180-187.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed as N-terminal His6-tagged ORN enzyme under control of the bacteriophage T7 promoter in Escherichia coli BL21(kDE3)/pLysS using derivatives of the expression vector pETMCSI Escherichia coli
expressed as N-terminal His6-tagged ORN enzyme under control of the bacteriophage T7 promoter in Escherichia coli BL21(kDE3)/pLysS using derivatives of the expression vector pETMCSI Homo sapiens
expressed as N-terminal His6-tagged ORN enzyme under control of the bacteriophage T7 promoter in Escherichia coli BL21(kDE3)/pLysS using derivatives of the expression vector pETMCSI Mycolicibacterium smegmatis

Inhibitors

Inhibitors Comment Organism Structure
Ni2+ effective competitive inhibitor Escherichia coli
Ni2+ effective competitive inhibitor Homo sapiens
Ni2+ effective competitive inhibitor Mycolicibacterium smegmatis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.27
-
5'-p-nitrophenyl-TMP in the presence of 3 mM Mn2+ Homo sapiens
0.28
-
5'-p-nitrophenyl-TMP in the presence of 3 mM Mn2+ Mycolicibacterium smegmatis
0.46
-
5'-p-nitrophenyl-TMP in the presence of 1 mM Mn2+ Mycolicibacterium smegmatis
0.54
-
5'-p-nitrophenyl-TMP in the presence of 3 mM Mn2+ Escherichia coli
0.85
-
5'-p-nitrophenyl-TMP in the presence of 1 mM Mn2+ Homo sapiens
1.9
-
5'-p-nitrophenyl-TMP in the presence of 1 mM Mn2+ Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
Escherichia coli
Mg2+
-
Homo sapiens
Mg2+
-
Mycolicibacterium smegmatis
Mn2+ preferred over Mg2+ Escherichia coli
Mn2+ preferred over Mg2+ Homo sapiens
Mn2+ preferred over Mg2+ Mycolicibacterium smegmatis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
21638
-
2 * 21638, electrospray ionisation mass spectrometry Escherichia coli
24576
-
2 * 24576, electrospray ionisation mass spectrometry Homo sapiens
24637
-
2 * 24637, electrospray ionisation mass spectrometry Mycolicibacterium smegmatis

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-
Homo sapiens
-
-
-
Mycolicibacterium smegmatis
-
-
-

Purification (Commentary)

Purification (Comment) Organism
on Ni-NTA column and by gel filtration, more than 95% pure Escherichia coli
on Ni-NTA column and by gel filtration, more than 95% pure Homo sapiens
on Ni-NTA column and by gel filtration, more than 95% pure Mycolicibacterium smegmatis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.77
-
purified enzyme Escherichia coli
7.1
-
purified enzyme Homo sapiens
14.9
-
purified enzyme Mycolicibacterium smegmatis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5'-p-nitrophenyl-TMP + H2O
-
Escherichia coli p-nitrophenol + TMP
-
?
5'-p-nitrophenyl-TMP + H2O
-
Homo sapiens p-nitrophenol + TMP
-
?
5'-p-nitrophenyl-TMP + H2O
-
Mycolicibacterium smegmatis p-nitrophenol + TMP
-
?

Subunits

Subunits Comment Organism
homodimer 2 * 21638, electrospray ionisation mass spectrometry Escherichia coli
homodimer 2 * 24576, electrospray ionisation mass spectrometry Homo sapiens
homodimer 2 * 24637, electrospray ionisation mass spectrometry Mycolicibacterium smegmatis

Synonyms

Synonyms Comment Organism
ORN
-
Escherichia coli
ORN
-
Homo sapiens
ORN
-
Mycolicibacterium smegmatis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
-
Escherichia coli
25
-
-
Homo sapiens
25
-
-
Mycolicibacterium smegmatis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.03 0.55 5'-p-nitrophenyl-TMP in the presence of 3 mM Mn2+ Homo sapiens
0.04 1.97 5'-p-nitrophenyl-TMP in the presence of 1 mM Mn2+ Homo sapiens
0.04 1.97 5'-p-nitrophenyl-TMP in the presence of 3 mM Mn2+ Mycolicibacterium smegmatis
0.057 0.65 5'-p-nitrophenyl-TMP in the presence of 1 mM Mn2+ Mycolicibacterium smegmatis
2.15
-
5'-p-nitrophenyl-TMP in the presence of 1 mM Mn2+ Escherichia coli
2.3
-
5'-p-nitrophenyl-TMP in the presence of 3 mM Mn2+ Escherichia coli
7.417
-
5'-p-nitrophenyl-TMP in the presence of 3 mM Mn2+ Homo sapiens
8.767
-
5'-p-nitrophenyl-TMP in the presence of 1 mM Mn2+ Homo sapiens
9.317
-
5'-p-nitrophenyl-TMP in the presence of 3 mM Mn2+ Mycolicibacterium smegmatis
10.88
-
5'-p-nitrophenyl-TMP in the presence of 1 mM Mn2+ Mycolicibacterium smegmatis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
-
Escherichia coli
8
-
-
Homo sapiens
8
-
-
Mycolicibacterium smegmatis

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.022
-
Ni2+
-
Homo sapiens
0.039
-
Ni2+
-
Escherichia coli
0.24
-
Ni2+
-
Mycolicibacterium smegmatis