A series of protein-polymer conjugates is contructed between FnCel5a cellulase and synthetic polymer created by reversible addition-fragmentation chain transfer (RAFT) polymerization, and tested using enzymatic activity assays and biophysical stability measures.. A grafting to strategy is employed to create the protein-polymer conjugates, with all polymers synthesized to have essentially the same molecular weight of ca. 30 units. These polymers vary in their functionality including strong/weak hydrogen bond donors/acceptors, anionic and cationic charges as pendant groups. Enzymatic activity was assesed by the hydrolysis of carboxymethyl. All cellulase-polymer conjugates are investigated for enzymatic activity compared to the native enzyme. All protein-polymer conjugates were found to have enhanced activity compared to the native enzyme, attributed to beneficial interactions between the polymer and the substrate. Protein stabiliy was assessed using biophysical and biochemical methods. Thermal stability was assessed using differential scanning fluorimetry, and the bioconjugates were found to have comparable thermal stability to the native enzyme. Resistance to denaturation by chemical challenges was assessed using a functional assay against organic solvent. Additionally, data suggests that functional groups that interact with the protein through non covalent hydrogen bonds or ionic interactions can lead to stabilization of the protein.

Enzyme-Polymer conjugates. Improving the activity of cellulase for biofuel applications

253rd National Meeting (2017)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.