Hydroxylamine → 5'-Adp

HRID 1893010Notes

Reaction equation

HRID 1893010 reaction equation
C(O)C(N)(CO)CO.Cl.[P:10]([O:22][CH2:23][C@H:24]1[O:28][C@@H:27]([N:29]2[C:38]3[N:37]=[CH:36][N:35]=[C:33]([NH2:34])[C:32]=3[N:31]=[CH:30]2)[C@H:26]([OH:39])[C@@H:25]1[OH:40])([O:13][P:14]([O:17]P(O)(O)=O)([OH:16])=[O:15])(=[O:12])[OH:11].NO.S([O-])([O-])(=O)=O.[NH4+].[NH4+].S([O-])([O-])(=O)=O.[Mg+2].[NH2:56][C@H:57]([C:62]([OH:64])=[O:63])[CH2:58][C:59]([OH:61])=[O:60]>O>[P:10](=[O:11])([OH:22])([OH:13])[OH:12].[NH2:56][C@H:57]([C:62]([OH:64])=[O:63])[CH2:58][C:59]([OH:61])=[O:60].[P:10]([O:22][CH2:23][C@H:24]1[O:28][C@@H:27]([N:29]2[C:38]3[N:37]=[CH:36][N:35]=[C:33]([NH2:34])[C:32]=3[N:31]=[CH:30]2)[C@H:26]([OH:39])[C@@H:25]1[OH:40])([O:13][P:14]([OH:16])([OH:17])=[O:15])(=[O:11])[OH:12]

Participants

9

Reactant 6

HCID 787HydroxylamineHCID 59575'-AtpHCID 5960L-Aspartic AcidHCID 24083Sulfuric acid magnesium salt (1:1)HCID 93573Tromethamine hydrochlorideHCID 6097028Ammonium Sulfate

Product 2

HCID 60225'-AdpHCID 44146787170082-99-2

Reagents, catalysts & solvents 1

HCID 962WaterSolvent

Procedure

The crude enzyme solution was pretreated at 30 to 80° C. for 1, 3, 5 or 10 minutes and reacted with a reaction mixture at 30° C. The reaction mixture was prepared by adding the crude enzyme solution to an aqueous solution containing 100 mM Tris-HCl (pH 7.5), 10 mM ATP (pH 7.5), 600 mM hydroxylamine, 600 mM ammonium sulfate, 10 mM magnesium sulfate, 50 mM L-aspartic acid (pH 7.5), and adjusted to the total volume of 500 μl with sterilized water. The reaction was allowed to proceed for a predetermined period of time, and the reaction was terminated by addition of 750 μl of a reaction terminating solution. The reaction terminating solution had a composition of 4% trichloroacetic acid, 10% FeCl2 and 1.4 N hydrochloric acid. The reaction mixture added with the reaction terminating solution was centrifuged at 15,000 rpm for 5 minutes, and the absorbance of the supernatant was measured at a wavelength of 540 nm. This measurement was performed for quantifying color development of hydroxymate produced by the reaction in which L-aspartic acid phosphoric acid salt, ADP and hydroxymate were produced from L-aspartic acid, ATP and hydroxylamine. The reaction mixture not containing L-aspartic acid was used as a blank.

Workup

  1. customwas pretreated at 30 to 80° C. for 1, 3, 5 or 10 minutes
  2. customreacted with a reaction mixture at 30° C
  3. customThe reaction mixture was prepared
  4. additionby adding the crude enzyme solution to an aqueous solution
  5. waitto proceed for a predetermined period of time
  6. customthe reaction was terminated by addition of 750 μl of a reaction
  7. customterminating solution
  8. customThe reaction terminating solution
  9. additionThe reaction mixture added with the reaction
  10. customterminating solution

Sources & evidence

ORD record
ord-d6eccbf131ec45bf83be37e392da3df7
Source dataset
uspto-grants-2013_08
Patent number
US08510053B2

Reaction notes

Loading…