N-Acetyl-D-Mannosamine → N-Acetyl-Neuraminic Acid

Participants

Reaction Steps

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%).

Provenance & Evidence