The synthesis of 4-methylumbelliferyl α-ketoside of N-acetylneuraminic acid and its use in a fluorometric assay for neuraminidase

Robert W. Myers, Reiko T. Lee, Yuan Chuan Lee, George H. Thomas, Linda W. Reynolds, Yoshihiro Uchida

Research output: Contribution to journalArticlepeer-review

Abstract

4-Methylumbelliferyl α-ketoside of N-acetylneuraminic acid was synthesized by reacting the sodium salt of 4-methylumbelliferone with the 2-chloro-2-deoxy derivative of peracetylated methyl N-acetylneuraminate, followed by preparative silica gel chromatography, deblocking, and purification by gel filtration on Sephadex G-25. The final product was isolated as either the sodium or ammonium salt, and its suitability as a substrate for neuraminidase was evaluated. The optimal pH values for various neuraminidases were 5.6 in acetate buffer (Arthrobacter ureafaciens), 5.0-5.1 in acetate buffer (Clostridium perfringens), and 4.4 in phosphate-citrate buffer (human fibroblasts). Km values for these enzymes at the optimal pH were 6 × 10-4 m (Arthrobacter), 1 × 10-4 m (Clostridium), and 3 × 10-4 m (human fibroblasts).

Original languageEnglish (US)
Pages (from-to)166-174
Number of pages9
JournalAnalytical biochemistry
Volume101
Issue number1
DOIs
StatePublished - Jan 1 1980

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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