반응 #663667

반응 방정식

N-acetylneuraminate

N-acetylmannosamine

sodium pyruvate

N-acetylmannosamine

sodium pyruvate

→

N-acetylneuraminic acid

수율 39.8%

용매

water

water

water

반응 조건

후처리

1. 1
   기타The enzyme bag was removed
2. 2
   기타reused for new batch of reaction
3. 3
   세척The resin was then washed with water (300 ml)
4. 4
   세척eluted with 0.5M HCOOH solution
5. 5
   기타The eluate was collected
6. 6
   기타vacuum evaporated to dryness
7. 7
   workup.DISSOLUTIONThe residue was dissolved in water (2 ml)
8. 8
   workup.ADDITIONthen diluted with glacial acetic acid (10 ml) at 4° C. overnight
9. 9
   여과filtered the crystals
10. 10
    세척washed with EtOH
11. 11
    기타dried

실험 절차

To a pH 7.0˜7.5 solution of N-acetylmannosamine (2.7 g, 12.2 mmole) and sodium pyruvate (2.7 g, 24.5 mmole) in water (15 ml) was added a dialysis bag (cut off MW of 20,000) containing N-acetylneuraminate lyase [EC 4.1.3.3] (25 units) in a reaction mixture of N-acetylmannosamine (0.54 g) and sodium pyruvate (0.54 g) in water (3 ml) at pH 7.0-7.5. The reaction mixture was shaken at 60 r.p.m. at 30° C. for 5 days. The enzyme bag was removed and reused for new batch of reaction. The reaction mixture was diluted with water (15 ml), then passed through a column of Amberlite IRA-400 (HCOO− form) (150 ml). The resin was then washed with water (300 ml), eluted with 0.5M HCOOH solution. The eluate was collected and vacuum evaporated to dryness. The residue was dissolved in water (2 ml) then diluted with glacial acetic acid (10 ml) at 4° C. overnight, filtered the crystals, washed with EtOH, dried to afford N-acetylneuraminic acid as white crystalline powder (1.5 g, 39.8%).

출처