Investigating catalytic diversity in copper radical oxidases for potential bioproduct applications
department or school: Science, Faculty of
department or school: Chemistry, Department of
data provider: DSpace
degree campus: UBCV
creator: Cleveland, Maria
issued: 2022-08-31T07:00:00Z
issued: 2021
related degree: Doctor of Philosophy - PhD
degree grantor: University of British Columbia
description: The utilization and valorization of biomass has been proposed as a sustainable alternative to petroleum-based energy, chemicals and materials. Microbial enzymes can be used as selective catalysts for the depolymerization and functionalization of plant material. In this regard, the copper radical oxidases (CROs) from Auxiliary Activity Family 5 subfamily 2 (AA5_2) are attractive targets since they oxidize primary alcohols in a chemo-selective manner to the corresponding aldehydes without utilizing complex and expensive organic cofactors. AA5_2 was first defined by the archetypal galactose oxidase from Fusarium graminearum, however, other fungal AA5_2 members have recently been shown to comprise a wide range of specificities for aromatic, aliphatic and furan-based alcohols. This suggests a broader substrate scope of CROs for biocatalytic applications. However, only 10% of the annotated AA5_2 members have been characterized to date. The work presented in this thesis describes the biochemical characterization of fourteen fungal copper radical oxidases, recombinantly produced in Pichia pastoris, encompassing galactose oxidases, raffinose oxidases, broad spectrum alcohol oxidases, non-carbohydrate alcohol oxidases and aryl alcohol oxidases. Detailed product analysis was conducted of carbohydrates, furans and glycerol oxidized by these novel enzymes to further explore their potential applications. Of note, we observed direct oxidation of 5-hydroxymethylfurfural (HMF), an important platform chemical derived from biomass, to 5-formyl-2-furoic acid (FFCA) by six enzymes. One enzyme facilitated the direct one-pot conversion of HMF to 2,5-furandicarboxylic acid (FDCA), a building block for bio-plastic production. Through docking of galactose into homology models of these newly characterized enzymes, and comparing it to the archetypal galactose oxidase, key amino acid residues important for substrate specificity have been proposed. Finally, the potential of enzyme immobilization…
aggregated CHO: https://circle.library.ubc.ca/rest/handle/2429/79415
note: INVESTIGATING CATALYTIC DIVERSITY IN COPPER RADICAL OXIDASES FOR POTENTIAL BIO-PRODUCT APPLICATIONS by Maria Cleveland B.Sc., Queens University, 2014 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY in THE FACULTY OF GRADUATE AND POSTDOCTORAL STUDIES (Chemistry) THE UNIVERSITY OF BRITISH COLUMBIA (Vancouver) August 2021 © Maria Cleveland, 2021 ii The following individuals certify that they have read, and recommend to the Faculty of Graduate and Postdoctoral Studies for acceptance, the dissertation entitled: INVESTIGATING CATALYTIC DIVERSITY IN COPPER RADICAL OXIDASES FOR POTENTIAL BIO-PRODUCT APPLICATIONS submitted by Maria Cleveland in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Chemistry Examining Committee: Dr. Harry Brumer, Professor, Chemistry, UBC Supervisor Dr. Stephen Withers, Professor, Chemistry, UBC Supervisory Committee Member Dr. David Perrin, Professor, Chemistry, UBC University Examiner Dr. Kirsten Wolthers, Associate Professor, Chemistry, UBC University Examiner Additional Supervisory Committee Members: Dr. Pierre Kennepohl, Associate Professor, Chemistry, UBC Supervisory Committee Member Dr. Mark MacLachlan, Professor, Chemistry, UBC Supervisory Committee Member iii Abstract The utilization and valorization of biomass has been proposed as a sustainable alternative to petroleum-based energy, chemicals and materials. Microbial enzymes can be used as selective catalysts for the depolymerization and functionalization of plant material. In this regard, the copper radical oxidases (CROs) from Auxiliary Activity Family 5 subfamily 2 (AA5_2) are attractive targets since they oxidize primary alcohols in a chemo-selective manner to the corresponding aldehydes without utilizing complex and expensive organic cofactors. AA5_2 was first defined by the archetypal galactose oxidase from Fusarium graminearum, however, other fungal AA5_2 members have recently been shown to compr…
has type: Thesis/Dissertation
date issued: 2021-11
is shown at: 10.14288/1.0401495
language: eng
degree discipline: Chemistry
provider: Vancouver : University of British Columbia Library
publisher: University of British Columbia
rights: Attribution-NonCommercial-NoDerivatives 4.0 International
rights URI: http://creativecommons.org/licenses/by-nc-nd/4.0/
scholar level: Graduate
contributor: Brumer, Harry
type: Text
identifier URI: http://hdl.handle.net/2429/79415