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N8264

Sigma-Aldrich

Nucleoside Phosphorylase from microorganisms

lyophilized powder, ≥10 units/mg protein

Sinonimo/i:

Nucleoside Phosphorylase bacterial, PNP, Purine nucleoside phosphorylase, Purine nucleoside:orthophosphate ribosyltransferase

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About This Item

Numero CAS:
Classificazione EC (Enzyme Commission):
Numero CE:
Numero MDL:
Codice UNSPSC:
12352204

Forma fisica

lyophilized powder

Attività specifica

≥10 units/mg protein

PM

~120 kDa

Composizione

Protein, 40-80%

Temperatura di conservazione

−20°C

InChI

1S/C10H12N4O4/c15-2-6-7(16)8(17)10(18-6)14-4-13-5-1-11-3-12-9(5)14/h1,3-4,6-8,10,15-17H,2H2/t6-,7-,8-,10-/m1/s1
MRWXACSTFXYYMV-FDDDBJFASA-N

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Descrizione generale

Nucleoside Phosphorylase are trimeric and have a molecular mass of about 100 kDa. Each monomer consists of a mixed β-sheet core flanked by eight α-helices and a purine binding in hydrophobic pocket.
Nucleoside phosphorylase is an enzyme important for the biosynthesis of nucleosides.

Applicazioni

Nucleoside Phosphorylase from microorganisms has been used:
  • in phosphopentomutase assay for adenosine synthesis
  • for the conversion of 7-methylthioguanosine to 7-methylthioguanine
  • in the synthesis of cytokinins in lyophilized cotton plant samples

Nucleoside phosphorylase is used in coupled enzyme systems to measure protein dephosphorylation.

Azioni biochim/fisiol

Nucleoside Phosphorylase catalyzes the synthesis of nucleoside derivatives.
This enzyme is useful for enzymatic determination of inorganic phosphorus, 5′-nucleotidase and adenosine deaminase when coupled with xanthine oxidase (XTO-212) and uricase (UAO-201, UAO-211). Purine nucleoside phosophorylase has shown the ability to perform both phosphorylosis and synthesis of purine deoxy- and ribonucleosides. It has also been found that membrane-associated nucleoside phosphorylases may have a transmembranal orientation with their base and ribose-1-P binding sites on opposite sides of the membrane.
Catalyzes the following reaction:purine nucleoside + phosphate = purine + alpha-D-ribose 1-phosphate

Proprietà fisiche

Isoelectric point : 4.1 +/- 0.1
Michaelis constants : 6.4 x 10-5M (Inosine), 3.2x10-4M (Pi)
Inhibitors : p-Chloromercuribenzoate, SDS, Hg++, Ag+Optimum pH : 7.5 - 8.0
Optimum temperature : 65oC
pH Stability : pH 6.0 - 9.0 (30oC, 16hr)
Thermal stability : below 60oC (pH 7.7, 30min)

Definizione di unità

One unit will cause the phosphorolysis of 1.0 μmole of inosine to hypoxanthine and ribose 1-phosphate per min at pH 7.4 at 25 °C.

Stato fisico

Lyophilized powder containing potassium gluconate, mannitol and EDTA

Codice della classe di stoccaggio

11 - Combustible Solids

Classe di pericolosità dell'acqua (WGK)

WGK 3

Punto d’infiammabilità (°F)

Not applicable

Punto d’infiammabilità (°C)

Not applicable

Dispositivi di protezione individuale

Eyeshields, Gloves, type N95 (US)


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Enzymatic synthesis of nucleosides by nucleoside phosphorylase co-expressed in Escherichia coli
Ding QB, et al.
Journal of Zhejiang University. Science. B, 11(11), 880-888 (2010)
A continuous kinetic assay for adenylation enzyme activity and inhibition
Wilson DJ and Aldrich CC
Analytical Biochemistry, 404(1), 56-63 (2010)
Measurement of nonribosomal peptide Synthetase Adenylation domain activity using a continuous hydroxylamine release assay
Duckworth BP, et al.
Methods in Molecular Biology, 53-61 (2016)
Structural basis for substrate specificity of Escherichia coli purine nucleoside phosphorylase
Bennett EM, et al.
The Journal of biological chemistry, 278(47), 47110-47118 (2003)
A simple assay for determining activities of phosphopentomutase from a hyperthermophilic bacterium Thermotoga maritima
Moustafa HMA, et al.
Analytical biochemistry, 501, 75-81 (2016)

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