反応 #663667

反応方程式

N-acetylneuraminate

N-acetylmannosamine

sodium pyruvate

N-acetylmannosamine

sodium pyruvate

→

N-acetylneuraminic acid

収率 39.8%

溶媒

water

water

water

反応条件

後処理

1. 1
   その他The enzyme bag was removed
2. 2
   その他reused for new batch of reaction
3. 3
   洗浄The resin was then washed with water (300 ml)
4. 4
   洗浄eluted with 0.5M HCOOH solution
5. 5
   その他The eluate was collected
6. 6
   その他vacuum 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
   ろ過filtered the crystals
10. 10
    洗浄washed with EtOH
11. 11
    その他dried

実験手順

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%).

出典