(723b) “Dock and Lock” Covalent Assembly of Designer Cellulosomes
AIChE Annual Meeting
2019
2019 AIChE Annual Meeting
Fuels and Petrochemicals Division
Developments in Alternative Fuels and Enabling Technologies II
Thursday, November 14, 2019 - 3:55pm to 4:20pm
In this study, we developed a âdock and lockâ method to enable irreversible assembly of cellulosomes by introducing a covalent bond to the cohesin-dockerin interaction. Specifically, homology modeling, docking, and structural analysis were performed to design cohesins and dockerins with single cysteine mutations so that cohesin-dockerin guided assembly causes the formation of a disulfide bond between the two proteins. Our data showed that introducing the cysteine mutations did not affect the cohesin-dockerin interaction, but created a covalent complex that has a greatly reduced dissociation rate and much improved stability without dissociation even under boiling conditions. As a result, these engineered cohesin-dockerin pairs improved the assembly efficiency of designer cellulosomes, which is further translated to enhanced catalytic properties. The method reported here provides a simple way to form covalent designer cellulosomes and can be seamlessly incorporated into any large protein assemblies with minimal design implications.