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Metabolic engineering for optimizing isobutanol production in Synechocystis PCC 6803
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Biology Education Centre.
2018 (English)Independent thesis Advanced level (degree of Master (Two Years)), 30 credits / 45 HE creditsStudent thesis
Abstract [en]

The diminishing of fossil fuels and growing concerns towards climate change have intensified biofuel production from renewable resources. Recently, progresses are made in microbial production of biofuels. Among various biofuels, isobutanol is gaining an increasing attention due to its high energy content and suitable chemical and physical properties, enabling it to be a suitable substitution of fossil fuel. In this study, instead of using heterotrophic microorganisms, we performed metabolic engineering of Synechocystis PCC 6803 (Synechocystis) for isobutanol production under autotrophic condition. After introduced 2-keto acid pathway, Synechocystis is able to produce isobutanol when provided with water, carbon dioxide and solar energy. When cultivated in an optimal condition (50 μmol photons m-1s-2 and adjusted pH to 7-8 with HCl), the engineered strain pEEK2-ST was able to produce 425 mg L-1 in-flask isobutanol titer and 911 mg L-1 cumulative isobutanol titer, respectively, in 46 days. There should be bottlenecks existing in 2-keto acid pathway based on the similar isobutanol production of strain pEEK2-ST with and without pyruvate addition. However, the attempt to identify potential bottlenecks of upstream genes by overexpressing ST and one of the three upstream genes failed, instead what we conclude is that the isobutanol production is tightly correlated to Kivd (ST) expression level. Thus, more strategies will be employed for identifying bottlenecks successfully and further improvement of isobutanol production in the future. In conclusion, this study demonstrates the importance of cultivation condition on isobutanol production in Synechocystis.

Place, publisher, year, edition, pages
2018. , p. 46
Keywords [en]
Synechocystis PCC 6803, isobutanol, metabolic engineering
National Category
Natural Sciences Microbiology
Identifiers
URN: urn:nbn:se:uu:diva-352324OAI: oai:DiVA.org:uu-352324DiVA, id: diva2:1212800
Educational program
Master Programme in Biology
Supervisors
Available from: 2018-06-04 Created: 2018-06-03 Last updated: 2018-06-11Bibliographically approved

Open Access in DiVA

The full text will be freely available from 2020-06-11 15:29
Available from 2020-06-11 15:29

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CiteExportLink to record
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