Assembly and characterisation of Diels-Alderase polymer conjugates for industrial biocatalysis

Abstract

The Diels-Alder reaction is a keystone reaction in synthetic chemistry. It is used extensively in the preparation of pharmaceuticals, crop protection agents and commodity chemicals. Diels-Alder biocatalysts offer expedient, green routes to a vast array of useful molecules, however, the development of industrially relevant Diels-Alderases has been hampered by their unsuitability for use at extremes of temperature, pressure and pH and their incompatibility with organic solvents. In this study a two-step polymer surfactant conjugation method has been developed, optimised and applied to the natural Diels-Alderase AbyU. This method involves the modification of protein surfaces through chemical super-charging, followed by polymer surfactant conjugation to form an amphipathic surfactant corona. The resulting AbyU-polymer conjugates exhibit increased tolerance to ‘harsh’ reaction conditions and enhanced catalytic efficiency versus the wild type enzyme. These data support the value of the outlined conjugation approach as a general method for improving biocatalyst tolerance to, and performance under, ‘harsh’ reaction conditions.

Date of Award	1 Oct 2019
Original language	English
Awarding Institution	The University of Bristol
Supervisor	Steven G Burston (Supervisor) & Paul R Race (Supervisor)