Expanding the selectivity and activity of galactose oxidase towards bioactive saccharide

Lead Research Organisation: University of Manchester
Department Name: Chemistry

Abstract

This project aims to generate a library of sugar oxidases with novel and highly specific substrate selectivity based on parent galactose oxidase enzymes. We will use a combination of homology searches of genomic databases (Prozomix Limited),directed evolution (The University of Manchester), high-throughput enzyme expression in bacterial hosts (Prozomix) and fast high-resolution analytical screening techniques (Bio-Shape Ltd and The University of Manchester) for enzyme discovery.

Background
Biocatalytic oxidation reactions have the potential to substitute many chemically catalysed oxidations in the pharmaceutical and fine chemical industry due to their superior regio- and stereoselectivity and low environmental impact.[1] Furthermore, oxidative enzymatic cross-linking of proteins is an important method to functionalising foods.[2] Particularly attractive tools for such bioengineering methods are carbohydrate modifying enzymes such as oxidases, which can be highly selective as demonstrated by the widely used glucose and galactose oxidases. Despite their plethora of applications, a surprisingly narrow range of oxidases in terms of substrate specificity is currently available, which highlights the need for carbohydrates-modifying enzymes with broader substrate scopes.[3] This project aims to address this gap in available biocatalysts by developing a range of oxidases with distinct and diverse substrates specificity towards common glycans including galactosides, mannosides, lactose, fructose, sialic acids and amino sugars. We will select for biocatalysts that can be easily produced in gram quantities and larger in bacterial expression systems and thus could find wide applications in biotechnology including modification and remodelling of oligo- and polysaccharides (lactose, starch, cellulose), glycolipids and glycoproteins.

Aims
This project aims to now generate a designer library of active, robust and more diverse oxidases using a combination of key methods provided by project p

Publications

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Studentship Projects

Project Reference Relationship Related To Start End Student Name
BB/R505109/1 01/10/2017 30/09/2021
1960946 Studentship BB/R505109/1 20/09/2017 30/09/2021 Ashley Mattey