Specific glycosidase activity isolated from a random phage display antibody library

G. N. Goud, O. Artsaenko, M. Bols, M. Sierks

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

Carbohydrates serve as key receptor sites in various cellular events such as viral attachment, tumor formation, and tissue inflammation. A potential route to control these events is to manipulate targeted carbohydrate structures in vivo using specifically designed glycohydrolases. Here we show that a stereospecific catalytic activity designed toward a particular sugar and linkage can be readily isolated from a phage display antibody library derived from a nonimmunized host. The activity was isolated using a transition-state analogue mimicking an α-glucosidasic linkage as antigen and showed a 20-fold specificity for that sugar and linkage. The DNA sequence, however, contains a large deletion in the antibody gene, which also changes the downstream reading frame, resulting in a translated sequence containing only 57 amino acids that has a predominantly hydrophobic amino terminal and a strongly hydrophilic carboxy terminal. The isolated catalytic activity has a strong Ph dependence, attributable to one or more of the numerous potentially charged groups in the carboxyl terminal. While the protein readily forms more stable multimers, the 7.3-Kd monomer represents by far the smallest glycosidase enzyme reported to date and can provide substantial new information toward understanding and modifying glycosidase activity.

Original languageEnglish (US)
Pages (from-to)197-202
Number of pages6
JournalBiotechnology Progress
Volume17
Issue number1
DOIs
StatePublished - Feb 22 2001
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology

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