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Metabolic engineering of lipid yeast using new genome editing techniques

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dc.contributor Aalto-yliopisto fi
dc.contributor Aalto University en
dc.contributor.advisor Vartiainen, Eija
dc.contributor.author Mehik, Emma
dc.date.accessioned 2017-03-16T06:29:07Z
dc.date.available 2017-03-16T06:29:07Z
dc.date.issued 2017-02-13
dc.identifier.uri https://aaltodoc.aalto.fi/handle/123456789/24835
dc.description.abstract In this work the aim was to produce medium chain dicarboxylic acids with metabolically engineered Y. lipolytica. The demand for sustainable raw materials is continuously growing and there is a need to find alternatives to petroleum based raw materials. In the future, the usage of microbes for production of different valuable chemicals is a growing field. One suitable organism is the oleaginous yeast Yarrowia lipolytica, which can be engineered for efficient and economical production of bio-based chemicals such as dicarboxylic acids. Dicarboxylic acids are useful for example as monomers for plastics such as nylon. To achieve this the metabolism of Y. lipolytica has to be engineered, which can be done with the genomic engineering tool CRISPR/Cas9, which is a globally growing method to make site specific gene deletions fast and efficiently. In this work a markerless CRISPR/Cas9 system was created for Y. lipolytica. Especially the expression of guide RNA (gRNA) under different promoters and in vitro gRNA was studied. As a result, deletions were successfully made with gRNA under a synthetic promoter SCR1 and Cas9 in the same plasmid, but the efficiency was not as high as expected. In this study, dicarboxylic acids were produced from glucose, with a genetically modified Y. lipolytica (deletion of β-oxidation and overproduction of ω-oxidation enzymes). Thioesterases were added to this modified strain to increase production of specifically medium chain length dicarboxylic acids, but these thioesterases did not make a difference to the product dicarboxylic acid chain length. As a result, we were able to produce long chain (16-C18) dicarboxylic acids directly from sugars. There is definitely potential in producing dicarboxylic acids from sugars, but further modifications to production strains are needed to make the process more efficient. fi
dc.format.extent 72+13
dc.language.iso eng en
dc.title Metabolic engineering of lipid yeast using new genome editing techniques fi
dc.type G2 Pro gradu, diplomityö fi
dc.contributor.school Kemian tekniikan korkeakoulu fi
dc.subject.keyword metabolic engineering fi
dc.subject.keyword oleaginous yeasts fi
dc.subject.keyword Yarrowia lipolytica fi
dc.subject.keyword dicarboxylic acids fi
dc.subject.keyword CRISPR/Cas9 fi
dc.subject.keyword free fatty acids fi
dc.identifier.urn URN:NBN:fi:aalto-201703163081
dc.programme.major Biotechnology fi
dc.programme.mcode KE3002 fi
dc.type.ontasot Master's thesis en
dc.type.ontasot Diplomityö fi
dc.contributor.supervisor Frey, Alexander
dc.programme Master's Programme in Chemical, Biochemical and Materials Engineering fi
dc.ethesisid Aalto 8192
dc.location P! fi

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