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master's thesis
Molekularno-genetička karakterizacija konstruiranih sojeva kvasaca Scheffersomyces stipitis i Kluyveromyces marxianus te optimiranje metode transformacije Scheffersomyces stipitis
Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology, 2022. urn:nbn:hr:159:443601
Horvat, Maja
University of Zagreb
Faculty of Food Technology and Biotechnology
Department of Biochemical Engineering
Laboratory for Biology and Microbial Genetics

Institutional repository: Repository of the Faculty of Food Technology and Biotechnology

Cite this document

Horvat, M. (2022). Molekularno-genetička karakterizacija konstruiranih sojeva kvasaca Scheffersomyces stipitis i Kluyveromyces marxianus te optimiranje metode transformacije Scheffersomyces stipitis (Master's thesis). Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology. Retrieved from https://urn.nsk.hr/urn:nbn:hr:159:443601

Horvat, Maja. "Molekularno-genetička karakterizacija konstruiranih sojeva kvasaca Scheffersomyces stipitis i Kluyveromyces marxianus te optimiranje metode transformacije Scheffersomyces stipitis." Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2022. https://urn.nsk.hr/urn:nbn:hr:159:443601

Horvat, Maja. "Molekularno-genetička karakterizacija konstruiranih sojeva kvasaca Scheffersomyces stipitis i Kluyveromyces marxianus te optimiranje metode transformacije Scheffersomyces stipitis." Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2022. https://urn.nsk.hr/urn:nbn:hr:159:443601

Horvat, M. (2022). 'Molekularno-genetička karakterizacija konstruiranih sojeva kvasaca Scheffersomyces stipitis i Kluyveromyces marxianus te optimiranje metode transformacije Scheffersomyces stipitis', Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, accessed 03 December 2024, https://urn.nsk.hr/urn:nbn:hr:159:443601

Horvat M. Molekularno-genetička karakterizacija konstruiranih sojeva kvasaca Scheffersomyces stipitis i Kluyveromyces marxianus te optimiranje metode transformacije Scheffersomyces stipitis [Master's thesis]. Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology; 2022 [cited 2024 December 03] Available at: https://urn.nsk.hr/urn:nbn:hr:159:443601

M. Horvat, "Molekularno-genetička karakterizacija konstruiranih sojeva kvasaca Scheffersomyces stipitis i Kluyveromyces marxianus te optimiranje metode transformacije Scheffersomyces stipitis", Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, Zagreb, 2022. Available at: https://urn.nsk.hr/urn:nbn:hr:159:443601

Metadata
TitleMolekularno-genetička karakterizacija konstruiranih sojeva kvasaca Scheffersomyces stipitis i Kluyveromyces marxianus te optimiranje metode transformacije Scheffersomyces stipitis
Title (english)Molecular-genetic characterization of engeneered Scheffersomyces stipitis and Kluyveromyces marxianus a yeast strains and optimization of the Scheffersomyces stipites transformation method
AuthorMaja Horvat
MentorMarina Svetec Miklenić (mentor)
Committee memberIgor Slivac (predsjednik povjerenstva)
Committee memberIvan Krešimir Svetec (član povjerenstva)
Committee memberRenata Teparić (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Food Technology and Biotechnology
(Department of Biochemical Engineering)
(Laboratory for Biology and Microbial Genetics)
Zagreb
Defense date and country2022-09-23, Croatia
Scientific / art field,
discipline and subdiscipline		BIOTECHNICAL SCIENCES
Biotechnology
Abstract
Nekonvencionalni kvasci Kluyveromyces marxianus i Scheffersomyces stipitis imaju značajan potencijal kao proizvodni organizmi u različitim bioprocesima. S. stipitis jedan je od kvasaca s najvišom sposobnošću fermentacije ksiloze u etanol, dok K. marxianus osim što je termofilan, brzo raste i može koristiti širok spektar različitih supstrata. Međutim u oba kvasca uvođenje ciljnih genetičkih modifikacija otežano je zbog česte ilegitimne (nehomologne) rekombinacije. Prvi cilj ovog rada bila je molekularno-genetička karakterizacija prethodno konstruiranih transformanata K. marxianus i S. stipites, koji proizvode L-laktat i 2,3-butanediol, metodom hibridizacije po Sothern-u. U kvascu S. stipitis istražena je mogućnost transformacije metodom elektroporacije i metodom protoplastiranja te utjecaj koncentracije ribonukleinskog kompleksa pri korištenju sustava CRISPR/Cas9. Dobiveni rezultati ukazuju na zaključak da prethodno razvijena metoda protoplastiranja nije pogodna za transformaciju kvasca S. stipitis te da koncentracija ribonukleinskog kompleksa značajno utječe na uspješnost transformacije elektroporacijom.
Abstract (english)
Non-conventional yeasts Kluyveromyces marxianus and Scheffersomyces stipitis have significant potential as production organisms in various bioprocesses. S. stipitis is one of the yeasts with the highest ability to ferment xylose into ethanol, while K. marxianus, in addition to being thermophilic, grows quickly and can use a wide range of different substrates. However, in both yeasts, the introduction of target genetic modifications is difficult due to frequent illegitimate (non-homologous) recombination. The first goal of this work was the molecular-genetic characterization of previously constructed transformants of K. marxianus and S. stipites, which produce L-lactate and 2,3-butanediol, using the Southern blotting hybridization method. In the yeast S. stipitis, the possibility of transformation by the electroporation method and the protoplasting method and the influence of the ribonucleic complex concentration when using the CRISPR/Cas9 system were investigated. The obtained results indicate the conclusion that the previously developed protoplasting method is not suitable for the transformation of the yeast S. stipitis and that the concentration of the ribonucleic complex significantly affects the success of the transformation by electroporation.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:159:443601
Study programmeTitle: Molecular Biotechnology
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra inženjer/inženjerka molekularne biotehnologije (magistar/magistra inženjer/inženjerka molekularne biotehnologije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Terms of useLicence Attribution-NoDerivatives 4.0 International (CC BY-ND 4.0) icon
Created on2023-03-01 11:00:19