Hydrochloric Acid → Hypoxanthine

HRID 504987Notes

Reaction equation

HRID 504987 reaction equation
[C@@H]1([N:10]2[C:19]3[N:18]=[CH:17][N:16]=[C:14]([OH:15])[C:13]=3[N:12]=[CH:11]2)O[C@H](CO)[C@@H](O)[C@H]1O.Cl>>[NH:16]1[C:14](=[O:15])[C:13]2[NH:12][CH:11]=[N:10][C:19]=2[N:18]=[CH:17]1

Participants

3

Reactant 2

HCID 313Hydrochloric AcidHCID 178521066(2R,3S,4R,5R)-2-(hydroxymethyl)-5-(6-hydroxypurin-9-yl)oxolane-3,4-diol

Product 1

HCID 135398638Hypoxanthine

Procedure

The activity of the UdP enzyme was determined in a phosphorolysis test by incubating for 5 minutes at 30° C. the soluble fraction (cell extract) obtained by sonication of a known amount of a suspension of the cell paste and by centrifugation of the homogenate in 100 mM-pH 7 phosphate buffer containing 60 mM of the uridine substrate. The enzyme reaction was blocked by acidification with 0.1N HCl; the suspension was filtered and analysed by RP-HPLC on a C18 column (Hypersyl 100; 4.6×250 mm), eluting under isocratic conditions with a mobile phase constituted by 0.02 M K2HPO4 in methanol-H2O (4:96 v/v) and adjusted to pH 4.5 with NH4OH. The amount of uracil formed in the reaction was determined by reference to a standard curve and the enzyme activity of the cell preparation was calculated in μmol uracil/min/g wet cell paste (units/g). The activity of the PNP enzyme was determined in a phosphorolysis test by incubating for 10 minutes at 30° C. the soluble fraction (cell extract) obtained by sonication of a known amount of a suspension of the cell paste and by centrifugation of the homogenate in 100 mM-pH 7 phosphate buffer containing 50 mM of the inosine substrate. The enzyme reaction was blocked by acidification with 0.1N HCl; the suspension was filtered and analysed by RP-HPLC on a C18 column (Hypersyl 100; 4.6×250 mm), eluting under isocratic conditions with a mobile phase constituted by 0.02 M K2HPO4 in methanol-H2O (4:96 v/v) and adjusted to pH 4.5 with NH4OH. The amount of hypoxanthine formed in the reaction was determined by reference to a standard curve and the enzyme activity of the cell preparation was calculated in μmol hypoxanthine/min/g wet cell paste (units/g).

Workup

  1. customThe enzyme reaction
  2. filtrationthe suspension was filtered
  3. washeluting under isocratic conditions with a mobile phase

Sources & evidence

ORD record
ord-9cdc0a6824cb49738fff69b41db4d96d
Source dataset
uspto-grants-2010_08
Patent number
US07781189B2

Reaction notes

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