Discovering and harnessing camptothecin hydroxylase enzymes in Camptotheca acuminata for chemoenzymatic synthesis of anticancer camptothecin derivatives
department or school: Science, Irving K. Barber Faculty of (Okanagan)
department or school: Chemistry, Department of (Okanagan)
data provider: DSpace
degree campus: UBCO
creator: Nguyen, Minh Tuan Anh
issued: 2022-09-07T07:00:00Z
issued: 2021
related degree: Master of Science - MSc
degree grantor: University of British Columbia
description: Since the 1990s, topotecan and irinotecan, two camptothecin derivatives, have been widely used as anticancer drugs owing to their phenomenal topoisomerase-I inhibitory activity. In the pharmaceutical industry, topotecan and irinotecan are chemically synthesized from the prodrug camptothecin, a quinoline alkaloid from Camptotheca acuminata, via 10-hydroxycamptothecin (10-HCPT), using toxic reagents and harsh conditions. This project aims to develop a chemoenzymatic approach for 10-HCPT production by exploring its biosynthetic pathway in C. acuminata. Multidisciplinary techniques, including bioinformatics, molecular cloning, and organic synthesis, were implemented to discover and characterize the plant metabolic enzymes. Two cytochrome P450 monooxygenases have been discovered that regio-selectively hydroxylated camptothecin at two different positions, C-10 and C-11 on the CPT scaffold. The high regio-selectivity and conversion rate (12 mg/L – biotransformation rate) afford the production of hydroxycamptothecins (10/11-HCPT) in the baker’s yeast Saccharomyces cerevisiae. By combining cytochrome P450 enzymes and chemo-catalysts sequentially in the chemoenzymatic process, 13 camptothecin derivatives including topotecan and irinotecan have been successfully synthesized. This research is a step forward in camptothecin biochemistry and leading finally to the economical and sustainable production of more active and water-soluble camptothecin derivatives.
aggregated CHO: https://circle.library.ubc.ca/rest/handle/2429/80154
note: DISCOVERING AND HARNESSING CAMPTOTHECIN HYDROXYLASE ENZYMES IN Camptotheca acuminata FOR CHEMOENZYMATIC SYNTHESIS OF ANTICANCER CAMPTOTHECIN DERIVATIVES by Minh Tuan Anh Nguyen B.Sc., Vietnam National University Ho Chi Minh City – University of Science, 2019 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE in THE COLLEGE OF GRADUATE STUDIES (Chemistry) THE UNIVERSITY OF BRITISH COLUMBIA (Okanagan) October 2021 © Minh Tuan Anh Nguyen, 2021 ii The following individuals certify that they have read, and recommend to the College of Graduate Studies for acceptance, a thesis/dissertation entitled: DISCOVERING AND HARNESSING CAMPTOTHECIN HYDROXYLASE ENZYMES IN Camptotheca acuminata FOR CHEMOENZYMATIC SYNTHESIS OF ANTICANCER CAMPTOTHECIN DERIVATIVES submitted by Minh Tuan Anh Nguyen in partial fulfillment of the requirements of the degree of Master of Science. Dr. Thu-Thuy Dang, Irving K. Barber Faculty of Science, University of British Columbia Supervisor Dr. Susan Murch, Irving K. Barber Faculty of Science, University of British Columbia Supervisory Committee Member Dr. Frederic Menard, Irving K. Barber Faculty of Science, University of British Columbia Supervisory Committee Member Dr. Michael Deyholos, Irving K. Barber Faculty of Science, University of British Columbia University Examiner iii Abstract Since the 1990s, topotecan and irinotecan, two camptothecin derivatives, have been widely used as anticancer drugs owing to their phenomenal topoisomerase-I inhibitory activity. In the pharmaceutical industry, topotecan and irinotecan are chemically synthesized from the prodrug camptothecin, a quinoline alkaloid from Camptotheca acuminata, via 10-hydroxycamptothecin (10-HCPT), using toxic reagents and harsh conditions. This project aims to develop a chemoenzymatic approach for 10-HCPT production by exploring its biosynthetic pathway in C. acuminata. Multidisciplinary techniques, including bioinformatics, molecular cloning, and…
has type: Thesis/Dissertation
date issued: 2021-11
is shown at: 10.14288/1.0402944
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: Dang, Thu Thuy
type: Text
identifier URI: http://hdl.handle.net/2429/80154