Polymer-mediated enzyme stabilization and drug delivery

This dissertation describes the application of protein engineering for enzyme stabilization utilizing bioconjugation approaches with polymer; also utilizing triblock-polymeric micelle for delivering hydrophobic anti-cancer drugs.A bioconjugation strategy was developed with functionalized polymer, Polyethyleneglycol (PEG) to stabilize haloalkane dehalogenase DhaA and its variants, using the intrinsic lysine residues and mutated Cysteine residues. Unfortunately, none of the lysine-mediated conjugates retained their activities in non-biological environments such as elevated temperature, organic-co-solvent and high ionic strength. However, interestingly the cysteine-mediated conjugates retained activities with respect to their unconjugated counterparts.Apart from enzyme stabilization, a library of biodegradable and bioresorbable triblock-co-polymer was developed and tested their efficacy to load a wide-range of hydrophobic anti-cancer drugs. The assembly was further converted to a ROS-sensitive smart one with the potential to do targeted drug delivery. To summarize, this dissertation presents an innovative concept for bioconjugation via two widely targeted amino acid residues: Lysine and Cysteine for the stabilization of DhaA in non-biological conditions, and explores the engineering of a synthetic polymeric library for both controlled and targeted drug delivery.