Potassium Phosphate, Tribasic → HCID 54758621, compound

HRID 355625Notes

Reaction equation

HRID 355625 reaction equation
C1O[C@H]2[C@@H](O)[C@H]([O:9][C@@H:10]3[C@@H:15]([OH:16])[C@H:14]([O:17][C@H:18]4[C@@H:22]5[C@@H:23]([OH:59])[C@H:24]([O:26][C@@H:27]6[C@@H:32]([OH:33])[C@H:31]([O:34][C@H]7[C@@H]8[C@@H](O)[C@H](O[C@@H]9[C@@H](O)C(O)O[C@H](CO)[C@@H]9O)O[C@H]7CO8)[O:30][C@H:29]([CH2:56][OH:57])[C@@H:28]6[OH:58])[O:25][C@H:19]4[CH2:20][O:21]5)[O:13][C@H:12]([CH2:60][OH:61])[C@@H:11]3[OH:62])O[C@@H]1[C@H]2O.P([O-])([O-])([O-])=O.[K+].[K+].[K+]>P([O-])([O-])([O-])=O>[CH2:20]1[O:21][C@H:22]2[C@H:23]([OH:59])[C@H:24]([O:26][C@@H:27]3[C@@H:32]([OH:33])[C@H:31]([OH:34])[O:30][C@H:29]([CH2:56][OH:57])[C@@H:28]3[OH:58])[O:25][C@@H:19]1[C@H:18]2[O:17][C@@H:14]1[O:13][C@H:12]([CH2:60][OH:61])[C@H:11]([OH:62])[C@H:10]([OH:9])[C@H:15]1[OH:16]

Participants

4

Reactant 2

HCID 62657Potassium Phosphate, TribasicHCID 90470557HCID 90470557, compound

Product 1

HCID 54758621HCID 54758621, compound

Reagents, catalysts & solvents 1

HCID 1061PhosphateSolvent

Procedure

An amount of 30 mg of neoagarohexaose was dissolved in 10 ml of phosphate buffer of pH 7.8. Into this solution, a solution of 0.1 ml of potassium phosphate buffer (pH 7.8) which dissolved 0.1 U of agarase 0107 isolated and purified from Vibrio sp. JT0107-L4 was added. The resultant solution was incubated at 30° C. for 3 hours, then a solution of 0.1 ml of phosphate buffer (pH 7.8) which dissolved 1.5 U of α-NAOS hydrolase isolated and purified from Vibrio sp. JT0107-L4 was added therein. The resultant solution was incubated at 30° C. for 5 hours. The reaction solution was applied to an activated carbon column (5 mmφ×50 mm), which was then washed with 1 ml of water and 1 ml of 10% ethanol followed by elution with 3 ml of 24% ethanol. The solvents of the 24% ethanol fraction were removed by distillation using a rotary evaporator. Thereafter, freeze-drying was applied and the weight was determined; the weight was 11.4 mg. This reaction product was analyzed by thin layer chromatography; thereby, a substance of Rf=0.3 (developing solvent; butanol:ethanol:water=3:1:1) was confirmed. In addition, the mass number of the reaction product was determined using a fast atom bombardment mass spectrometer and found to be 486. From these facts, the substance was identified as agarotriose. The yield of the resulting agarotriose was calculated as 76%. The structure of agarotriose is shown below. ##STR2##

Workup

  1. customisolated
  2. custompurified from Vibrio sp
  3. additionJT0107-L4 was added
  4. dissolutiona solution of 0.1 ml of phosphate buffer (pH 7.8) which dissolved 1.5 U of α-NAOS
  5. customhydrolase isolated
  6. custompurified from Vibrio sp
  7. additionJT0107-L4 was added
  8. waitThe resultant solution was incubated at 30° C. for 5 hours
  9. washwas then washed with 1 ml of water and 1 ml of 10% ethanol
  10. washfollowed by elution with 3 ml of 24% ethanol
  11. customThe solvents of the 24% ethanol fraction were removed by distillation
  12. customa rotary evaporator
  13. customThereafter, freeze-drying

Sources & evidence

ORD record
ord-21b50c5e4f3c4336ae165911daf8b960
Source dataset
uspto-grants-1998_11
Patent number
US05834257

Reaction notes

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