Engineering montbretin A biosynthesis in Nicotiana benthamiana
department or school: Science, Faculty of
data provider: DSpace
degree campus: UBCV
creator: Sunstrum, Frederick Guy
issued: 2021-12-07T18:28:00Z
issued: 2021
related degree: Master of Science - MSc
degree grantor: University of British Columbia
description: The plant kingdom has always been a useful source of therapeutic molecules for humans. Montbretin A (MbA) is a complex acylated flavonol glycoside found in the underground storage organs, called corms, of the ornamental plant montbretia (Crocosmia x crocosmiiflora). MbA was discovered as a highly efficient and specific inhibitor of the human pancreatic α-amylase (HPA), making it an attractive candidate for the treatment of type 2 diabetes, a worldwide epidemic. The original plant source montbretia is not adapted for large scale cultivation, and MbA’s complex structure renders chemical synthesis infeasible for scalable production. Therefore, we are aiming to engineer a heterologous system for large-scale sustainable MbA production for treatment of type 2 diabetes. To this end we are using synthetic biology to develop an MbA producing Nicotiana benthamiana (tobacco) strain. This is possible due to the recent complete elucidation of the MbA biosynthetic pathway, including all essential genes and enzymes from montbretia. Small amounts of MbA can be produced in N. benthamiana through transient expression of the MbA biosynthetic enzymes. However, much higher quantities of montbretin B (MbB) and montbretin C (MbC), which differ from MbA in the identity of their acyl groups, were produced. MbB and MbC possess a coumaroyl and a feruloyl group, respectively, and are not effective HPA inhibitors. To influence acyl group availability, specifically increase the amount of caffeoyl-CoA for MbA biosynthesis, we investigated a set of enzymes involved in caffeoyl-CoA biosynthesis in montbretia, specifically 4-coumaroyl CoA ligases (4CL), hydroxycinnamoyl- CoA shikimate/quinate hydroxycinnamoyltransferases (HCT), 4-coumarate 3-hydroxylases (C3Hs), and caffeoyl-shikimate esterases (CSE). Co-expression of those caffeoyl-CoA biosynthetic genes together with MbA biosynthetic genes resulted in up to 10-fold higher MbA yields.
aggregated CHO: https://circle.library.ubc.ca/rest/handle/2429/80382
note: ENGINEERING MONTBRETIN A BIOSYNTHESIS IN NICOTIANA BENTHAMIANA by Frederick Guy Sunstrum B.Sc., The University of Ottawa, 2019 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE in THE FACULTY OF GRADUATE AND POSTDOCTORAL STUDIES (Genome Science and Technology) THE UNIVERSITY OF BRITISH COLUMBIA (Vancouver) November 2021 © Frederick Guy Sunstrum, 2021 ii The following individuals certify that they have read, and recommend to the Faculty of Graduate and Postdoctoral Studies for acceptance, a thesis entitled: Engineering Montbretin A Biosynthesis in Nicotiana benthamiana submitted by Frederick Guy Sunstrum in partial fulfillment of the requirements for the degree of Master of Science in Genome Science and Technology Examining Committee: Dr. Jörg Bohlmann, Professor, Michael Smith Laboratories, UBC Supervisor Dr. Stephen Withers, Professor, Michael Smith Laboratories, UBC Supervisory Committee Member Dr. Vikramaditya Yadav, Professor, Chemical and Biological Engineering, UBC Supervisory Committee Member Dr. Robert Guy, Professor, Forestry, UBC Additional Examiner iii Abstract The plant kingdom has always been a useful source of therapeutic molecules for humans. Montbretin A (MbA) is a complex acylated flavonol glycoside found in the underground storage organs, called corms, of the ornamental plant montbretia (Crocosmia x crocosmiiflora). MbA was discovered as a highly efficient and specific inhibitor of the human pancreatic α-amylase (HPA), making it an attractive candidate for the treatment of type 2 diabetes, a worldwide epidemic. The original plant source montbretia is not adapted for large scale cultivation, and MbA’s complex structure renders chemical synthesis infeasible for scalable production. Therefore, we are aiming to engineer a heterologous system for large-scale sustainable MbA production for treatment of type 2 diabetes. To this end we are using synthetic biology to develop an MbA producing Nicotiana benthami…
has type: Thesis/Dissertation
date issued: 2022-05
is shown at: 10.14288/1.0404494
language: eng
degree discipline: Genome Science and Technology
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: Bohlmann, Jörg
type: Text
identifier URI: http://hdl.handle.net/2429/80382