X-Neu5Ac: A novel substrate for chromogenic assay of neuraminidase activity in bacterial expression systems

Ikuo Fujii, Yoshiharu Iwabuchi, Tadashi Teshima, Tetsuo Shiba, Masakazu Kikuchi

Research output: Contribution to journalArticlepeer-review

32 Citations (Scopus)

Abstract

A chromogenic substrate 1, 5-bromo-4-chloroindol-3-yl 5-acetamido-3,5-dideoxy-α-d-glycero-d-galacto-2-nonulopyranosidoni c acid (X-Neu5Ac), has been synthesized to facilitate the screening of bacterial colonies or plaques for the detection of either natural or mutant neuraminidase activity. Substrate 1 was hydrolyzed by neuraminidase isolated from Clostridium perfringens to release a halogenated indol-3-ol 2 that undergoes rapid aerobic oxidation to form the dark blue pigment, 5,5′-dibromo-4,-4′-dichloroindigo 3. Preliminary kinetic studies indicate that this compound is a good substrate (Km 0.89 × 10-3 M) for neuraminidase and is quite stable under identical conditions in the absence of enzyme. These results suggest that X-Neu5Ac 1 can be useful to screen for bacterially-encoded enzyme production directly on agar plates.

Original languageEnglish
Pages (from-to)147-149
Number of pages3
JournalBioorganic and Medicinal Chemistry
Volume1
Issue number2
DOIs
Publication statusPublished - 1993 Aug
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmaceutical Science
  • Drug Discovery
  • Clinical Biochemistry
  • Organic Chemistry

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