Lectin Agglutinated Multienzyme Catalyst with Enhanced Substrate Affinity and Activity

Abstract

A method based on biological recognition was proposed to prepare cross-linked multiple glycoenzyme aggregates. With Concanavalin A (ConA) as a molecular glue, horseradish peroxidase (HRP) and glucose oxidase (GOx) were agglutinated and then cross-linked by glutaraldehyde, forming a GOx-ConA-HRP catalyst. The affinity of ConA to glucose enhanced the uptake of the substrate, reducing the Km of cross-linked GOx-ConA-HRP aggregates for glucose from 51 mM to 8.8 mM. The colocalization and clustering of cascade enzymes at nanoscale facilitated the intermediate consumption. These effects significantly improved the catalytic performance of the GOx-HRP cascade with a 1.5-fold increased specificity constant. The use of ConA as a molecular glue provides a facile way to construct a multienzyme catalyst with enhanced stability and activity.