Genetička transformacija kvasca Spathaspora passalidarum

undergraduate thesis

• Title (english): Genetic Transformation of the Yeast Spathaspora passalidarum
• Author: Lea Smolec
• Mentor: Ivan-Krešimir Svetec (mentor)
• Mentor: Angela Matanović (pomoć pri izradi, neposredni voditelj)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for Biology and Microbial Genetics); Zagreb
• Defense date and country: 2024-09-10, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Nekonvencionalni kvasci su sve vrste kvasaca izuzev Saccharomyces cerevisiae i Schizosaccharomyces pombe, dvije vrste koje se najčešće koriste u industriji. Mnogi nekonvencionalni kvasci imaju sposobnost metaboliziranja širokog spektra supstrata, proizvodnje specifičnih relevantnih sekundarnih metabolita, te često pokazuju veću otpornost na stresne uvijete kao što su visoke temperature, osmotski pritisak, prisutnost toksičnih tvari ili ekstremne pH vrijednosti. Jedna od biotehnološki najznačajnijih karakteristika nekonvencionalnog kvasca Spathaspora passalidarum je da zahvaljujući specifičnim genima i biosintetskim putevima može fermentirati ksilozu, šećer prisutan u lignoceluloznoj biomasi. Kako bi se iskoristio potencijal nekonvencionalnih kvasaca te unaprijedile njihove karakteristike, neophodno je moći ih genetički modificirati. Stoga, je cilj ovog rada bio transformirati kvasac Spathaspora passalidarum sa različitim transformirajućim molekulama DNA, te ispitati aktivnost ishodišta replikacije iz različitih vrsta kvasaca korištenjem plazmidnih vektora. Postupak transformacije proveden je metodom elektroporacije koja je prethodno razvijena za kvasac Brettanomyces bruxellensis, a dobiveni rezultati pokazali su da se najviša efikasnost transformacije postiže sa plazmidom koji sadrži ishodište replikacije iz kvasca Scheffersomyces stipitis.

Abstract (english)

Nonconventional yeasts include all yeast species except Saccharomyces cerevisiae and Schizosaccharomyces pombe, the two most commonly used species in industry. Many nonconventional yeasts have the ability to metabolize a wide range of substrates, produce specific relevant secondary metabolites, and often show greater resistance to stressful conditions such as high temperatures, osmotic pressure, the presence of toxic substances or extreme pH values. One of the most biotechnologically significant characteristics of the nonconventional yeast Spathaspora passalidarum is that, thanks to specific genes and biosynthetic pathways, it can ferment xylose, the sugar present in lignocellulosic biomass. To harness the potential of nonconventional yeasts and improve their characteristics, it is essential to be able to genetically modify them. Therefore, the aim of this work is to transform the yeast Spathaspora passalidarum with different transforming DNA molecules and to examine the activity of replication origins from various yeast species using plasmid vectors. The transformation process was carried out using the electroporation method previously developed for the yeast Brettanomyces bruxellensis, and the obtained results showed that the highest transformation efficiency is achieved with the plasmid containing the replication origin from the yeast Scheffersomyces stipitis.

• Keywords: nekonvencionalni kvasci; kvasac Spathaspora passalidarum; genetička transformacija; elektroporacija; plazmidni vektori
• Keywords (english): unconventional yeasts; Spathaspora passalidarum yeast; genetic transformation; electroporation; plasmid vectors
• Language: croatian
• URN:NBN: urn:nbn:hr:159:664227
• Study programme: Title: Biotechnology; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-09-26 08:42:10