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N8264

Sigma-Aldrich

Nucleoside Phosphorylase from microorganisms

lyophilized powder, ≥10 units/mg protein

Sinônimo(s):

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

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About This Item

Número CAS:
Número da licença da enzima:
Número CE:
Número MDL:
Código UNSPSC:
12352204
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Formulário

lyophilized powder

atividade específica

≥10 units/mg protein

peso molecular

~120 kDa

composição

Protein, 40-80%

temperatura de armazenamento

−20°C

cadeia de caracteres SMILES

[n]2(c3ncncc3nc2)[C@@H]1O[C@@H]([C@H]([C@H]1O)O)CO

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

chave InChI

MRWXACSTFXYYMV-FDDDBJFASA-N

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Descrição geral

Nucleoside Phosphorylase are trimeric and have a molecular mass of about 100 kDa.[1] Each monomer consists of a mixed β-sheet core flanked by eight α-helices and a purine binding in hydrophobic pocket.[1]
Nucleoside phosphorylase is an enzyme important for the biosynthesis of nucleosides. [2]

Aplicação

Nucleoside Phosphorylase from microorganisms has been used:
  • in phosphopentomutase assay for adenosine synthesis[3]
  • for the conversion of 7-methylthioguanosine to 7-methylthioguanine[4][5]
  • in the synthesis of cytokinins in lyophilized cotton plant samples[6]

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

Ações bioquímicas/fisiológicas

Catalyzes the following reaction:purine nucleoside + phosphate = purine + alpha-D-ribose 1-phosphate
Nucleoside Phosphorylase catalyzes the synthesis of nucleoside derivatives.[7]
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. [8] 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. [9]

propriedades físicas

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)

Definição da unidade

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.

forma física

Lyophilized powder containing potassium gluconate, mannitol and EDTA

Código de classe de armazenamento

11 - Combustible Solids

Classe de risco de água (WGK)

WGK 3

Ponto de fulgor (°F)

Not applicable

Ponto de fulgor (°C)

Not applicable

Equipamento de proteção individual

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)
A continuous kinetic assay for adenylation enzyme activity and inhibition
Wilson DJ and Aldrich CC
Analytical Biochemistry, 404(1), 56-63 (2010)
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)
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
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)

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