Collaborative self-assembled enzymes and self-immobilised biofilms for enhanced biocatalytic processes

Continuous production of bio-based products remains a significant challenge in biocatalysis. In this study, this issue is addressed by developing a novel one-step biocatalytic platform. This method leverages co-immobilization technology, which combines the efficient display of enzymes on the cell surface with the properties of cell biofilms immobilized on a carrier. This facilitates continuous catalytic production of biochemical products by surface-displayed enzymes. The surface-displayed pullulanase achieved efficient conversion in 16 batches, maintaining a conversion rate above 99% when pullulan was used as the substrate, corresponding to a space-time hydrolysis rate of 20 g/L/d. When natural starch was used as the substrate, resistant starch type 3 was successfully obtained. This study proposes a novel solution to address challenges encountered during multi-batch catalysis of pullulanase, presenting a highly promising strategy for continuous biological catalysis in a controlled environment.