Skip to content

Computational design, construction, and characterisation of artificial peroxidases

Bristol student theses: Doctoral ThesisDoctor of Philosophy (PhD)

Authors

  • Jonathan Jenkins

Research units

Abstract

One of the ambitious goals of synthetic biology is to produce biologically compatible catalysts that are cheap, robust, and sustainable. Tailor-made synthetic proteins that can mimic the highly specific and efficient catalysis of natural proteins are key to realising this goal for industrial biotechnology. Oxidoreductases are a diverse class of enzymes that catalyse a similarly diverse array of redox reactions, everything from electron transfer in photosynthesis to site-specific hydrocarbon oxidation in the mammalian liver. Sophisticated oxidoreductase functionality has long been the goal of the maquette protein design approach. The maquette approach aims to understand the necessary sequence structure relationship of these enzymes by iteratively designing in functionality into a non-functional de novo scaffold. This bottom up approach to enzyme design aims to produce biocompatible catalysts, desired by the biotechnology industry, whilst simultaneously providing an acid test for our understanding of oxidoreductase enzymology, by building an oxidoreductase from scratch.
This project focuses on c-type heme binding maquettes that have been designed to mimic the functionality of peroxidases en route to the more commercially interesting oxygenase or peroxygenase chemistry. These c-type heme binding maquettes have a de novo protein sequence which only contains the essential information for folding into a four-helix bundle that can covalently bind c-type heme. This project describes the last design iteration in producing a mono-histidine heme ligation in a c-type heme maquette known as C45. C45 functions as a catalytically efficient peroxidase and represents a resounding success of the maquette design principles.

Details

Original languageEnglish
Awarding Institution
Supervisors/Advisors
Award date19 Mar 2019

    Research areas

  • de novo, enzyme, design, maquette, peroxidase

Documents

View research connections

Research Outputs