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Exploiting SCRaMbLE to increase fatty acid synthesis in yeast

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thesis
posted on 28.03.2022, 22:01 by Thomas Robert Collier
Fatty acids (FAs) and lipids produced in yeast could provide an alternative source for consumable oils in the future. In this work, four recombinant yeast strains were developed using The SCRaMbLEing technology (Synthetic Chromosome Recombination and Modification by Lox-P Mediated Evolution) developed in the global Yeast 2.0 project aiming to build a completely synthetic Saccharomyces cerevisiae. The genes: ACC1 (acetyl-CoA carboxylase 1), ACS1 (acetyl-CoA synthetase), FAS1 (fatty acid synthase 1) and FAS2 (fatty acid synthase 2) enclosed by loxP sites were overexpressed in a semi-synthetic yeast strain equipped with a synthetic chromosome 14 containing about 260 Lox-P sites to enable SCRaMbLE. The lipid stain Nile red stain for neutral lipids was successfully used in conjunction with flow cytometry to develop a high through put screening protocol for yeast strains with increased FA content. The FA profiles of the SCRaMbLEd strains Syn-sXIV,Syn-sACC1, and Syns-FAS2 were then analysed by gas chromatography mass spectrometry and an increase in C16:0, C16:1, C18:0 and C18:1 FA content was detected in strains expressing ACC1, and FAS2.

History

Table of Contents

Chapter 1. Introduction -- Chapter 2. Materials and methods -- Chapter 3. Results and discussion -- Chapter 4. Conclusion -- References.

Notes

Bibliography: pages 50-54 Empirical thesis.

Awarding Institution

Macquarie University

Degree Type

Thesis MRes

Degree

MRes, Macquarie University, Faculty of Science and Engineering, Department of Molecular Sciences

Department, Centre or School

Department of Molecular Sciences

Year of Award

2018

Principal Supervisor

Helena Nevalainen

Additional Supervisor 1

Hugh Douglas Goold

Additional Supervisor 2

Angela Sun

Rights

Copyright Thomas Robert Collier 2018. Copyright disclaimer: http://mq.edu.au/library/copyright

Language

English

Extent

1 online resource (54 pages) diagrams, graphs

Former Identifiers

mq:71032 http://hdl.handle.net/1959.14/1270155