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Literature summary for 2.4.2.28 extracted from

  • Cacciapuoti, G.; Fusco, S.; Caiazzo, N.; Zappia, V.; Porcelli, M.
    Heterologous expression of 5'-methylthioadenosine phosphorylase from the archaeon Sulfolobus solfataricus: characterization of the recombinant protein and involvement of disulfide bonds in thermophilicity and thermostability (1999), Protein Expr. Purif., 16, 125-135.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression at a high level in Escherichia coli Saccharolobus solfataricus
overexpression in Escherichia coli strain RB791, amino acid determination, incorrect positioning of disulfide bonds, the recombinant enzyme is less thermostable and thermophilic than the native enzyme Saccharolobus solfataricus

Inhibitors

Inhibitors Comment Organism Structure
guanidine hydrochloride only recombinant enzyme Saccharolobus solfataricus
iodoacetamide only recombinant enzyme Saccharolobus solfataricus
iodoacetate only recombinant enzyme Saccharolobus solfataricus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.024
-
5'-methylthioadenosine recombinant enzyme Saccharolobus solfataricus
0.024
-
5'-methylthioadenosine wild-type and recombinant enzyme Saccharolobus solfataricus
0.125
-
phosphate recombinant enzyme Saccharolobus solfataricus
0.125
-
phosphate wild-type and recombinant enzyme Saccharolobus solfataricus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
27000
-
6 * 27000, recombinant enzyme, SDS-PAGE Saccharolobus solfataricus
160000
-
gel filtration Saccharolobus solfataricus
160000
-
recombinant enzyme, gel filtration Saccharolobus solfataricus
160000
-
recombinant enzyme Saccharolobus solfataricus

Organism

Organism UniProt Comment Textmining
Saccharolobus solfataricus
-
-
-
Saccharolobus solfataricus P50389
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme Saccharolobus solfataricus
recombinant from Escherichia coli Saccharolobus solfataricus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.115
-
-
Saccharolobus solfataricus
2.12
-
purified recombinant enzyme Saccharolobus solfataricus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5'-methylthioadenosine + phosphate
-
Saccharolobus solfataricus adenine + 5-methylthio-D-ribose 1-phosphate
-
?

Subunits

Subunits Comment Organism
hexamer 6 * 27000, recombinant enzyme, SDS-PAGE Saccharolobus solfataricus

Synonyms

Synonyms Comment Organism
5'-methylthioadenosine phosphorylase
-
Saccharolobus solfataricus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
70
-
assay at Saccharolobus solfataricus
100
-
recombinant enzyme Saccharolobus solfataricus
120
-
wild-type enzyme Saccharolobus solfataricus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30 120 recombinant enzyme Saccharolobus solfataricus
60 120 60°C: about 40% of maximal activity, 120°C: about 40% of maximal activity, recombinant enzyme Saccharolobus solfataricus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
the recombinant enzyme, expressed in Escherichia coli, is less thermostable and thermophilic than the native enzyme due to incorrect positioning of disulfide bonds Saccharolobus solfataricus
additional information
-
the recombinant 5'-methylthioadenosine phosphorylase is less thermophilic and thermostable than the Sulfolobus solfataricus enzyme, since an incorrect positioning of disulfide bonds within the molecule generates structures less stable to thermal unfolding Saccharolobus solfataricus
70
-
recombinant enzyme, 0.8 M DTT, stable Saccharolobus solfataricus
90
-
recombinant enzyme, 0.8 M DTT, 2 h, loss of 38% activity Saccharolobus solfataricus
100
-
recombinant enzyme, 1 h, 85% remaining activity Saccharolobus solfataricus
100
-
1 h, 15% loss of activity of recombinant enzyme, wild-type enzyme remains stable Saccharolobus solfataricus
111
-
recombinant enzyme, melting temperature Saccharolobus solfataricus
111
-
melting temperature of recombinant enzyme Saccharolobus solfataricus
118
-
recombinant enzyme in presence of 100 mM phosphate, melting temperature Saccharolobus solfataricus