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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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Visite la Librería de documentos

R L Rader et al.
Journal of bacteriology, 128(1), 290-301 (1976-10-01)
Although uridine and adenosine are converted by membrane-associated nucleoside phosphorylases to ribose-1-phosphate (ribose-1-P) and the corresponding bases (uracil and adenine), only ribose -1-P is accumulated within Salmonella typhimurium LT2 membrane vesicles. In accordance with these observations, no uptake is observed
Nutrient and hormone levels in cotton ovules during embryony
Fuller RJ, et al.
Plant Cell, Tissue and Organ Culture, 99(2), 183-192 (2009)
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)
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 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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