Pyruvate → N-Acetyl-Neuraminic Acid

Participants

Reaction Steps

The second reaction was run with the recovered enzyme under the same conditions with exceptions in the amounts of substrates (40 mmol of ManNAc and 400 mmol of pyruvate) and the volume of solution (260 mL). The 10-day operation gave 91% conversion of ManNAc to Neu5Ac. The recovered enzyme retained 50% of its initial activity, and the products in solution were stored at -10° C. until further purified. The excess pyruvate was removed three step procedure. Fifty-two mL portions of the solution were passed through a 2.5×40 cm column of Dowex 50W-X8 (hydrogen form) with water elution. The portion of the eluent which contained the product was lyophilized to a paste. The paste was slurried in ethyl acetate and the Neu5Ac product recovered by filtration. The collected white powder was dried under a stream of dry air to yield 2.7 g of Neu5Ac as a mono-hydrate. The 1H-NMR spectrum of the product so obtained was identical to the reported spectrum. Brown, E. B., Brey, W. S.,Jr., Weltner, W.,Jr. Biochem. Biophys. Acta 399 (1975) 124-30.

Provenance & Evidence