Compartmentalized co-immobilization of cellulase and cellobiose phosphorylase within zeolitic imidazolate framework efficiently synthesizes 1-p-Glc: Glycosylation of 18FDG

Enzymatic glycosylation is an efficient and biocompatible approach to enhance natural product bioavailability. Cellobiose phosphorylase, a novel glycosyltransferase, utilizes 1-phospho-glucose (1-p-Glc) as a glycosyl donor for regioselective glycosylation of various natural substrates. However, the high cost of 1-p-Glc limits the economic feasibility of the process. Thus, a dual-enzyme cascade system involving cellulase Ac Cel9A and cellobiose phosphorylase Cb CBP using a co-immobilization strategy was developed to overcome this challenge. The system utilizes low-cost carboxymethyl cellulose (CMC) for continuous 1-p-Glc production, which was then used in the fluorodeoxy glucose (FDG) glycosylation to generate fluorodeoxy cellobiose (FDC), which potentially traces fungal infections. The compartmentalized co-immobilization of the two enzymes within the internal and external regions of a porous zeolitic imidazolate framework-8 (ZIF-8) carrier enhanced the overall stability of the dual-enzyme system. The immobilized enzymes retained approximately 63.3 % activity after seven reuse cycles and 74 % catalytic efficiency after 12 days of storage at room temperature. Therefore, the developed co-immobilized multi-enzyme system holds significant potential for industrial biocatalysis applications.