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Merck

N8264

Sigma-Aldrich

Nucleoside Phosphorylase from microorganisms

lyophilized powder, ≥10 units/mg protein

Sinónimos:

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

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About This Item

Número de CAS:
Comisión internacional de enzimas:
EC Number:
MDL number:
UNSPSC Code:
12352204

form

lyophilized powder

specific activity

≥10 units/mg protein

mol wt

~120 kDa

composition

Protein, 40-80%

storage temp.

−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

InChI key

MRWXACSTFXYYMV-FDDDBJFASA-N

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General description

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.

Application

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.

Biochem/physiol Actions

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

Physical properties

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)

Unit Definition

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.

Physical form

Lyophilized powder containing potassium gluconate, mannitol and EDTA

Storage Class

11 - Combustible Solids

wgk_germany

WGK 3

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

Eyeshields, Gloves, type N95 (US)


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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)
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)
A simple assay for determining activities of phosphopentomutase from a hyperthermophilic bacterium Thermotoga maritima
Moustafa HMA, et al.
Analytical biochemistry, 501, 75-81 (2016)
Simone Kunzelmann
Methods in molecular biology (Clifton, N.J.), 2263, 289-318 (2021-04-21)
Assays for the detection of inorganic phosphate (Pi) are widely used to measure the activity of nucleotide hydrolyzing enzymes, such as ATPases and GTPases. The fluorescent biosensors for Pi, described here, are based on fluorescently labeled versions of E. coli

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