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master's thesis
Konstrukcija sojeva Saccharomyces cerevisiae za produkciju aminokiselina nalik mikosporinu
Zagreb: University of Zagreb, Faculty of Science, 2019. urn:nbn:hr:217:565236
Jurić, Vanja
University of Zagreb
Faculty of Science
Department of Biology

Institutional repository: Repository of the Faculty of Science

Cite this document

Jurić, V. (2019). Konstrukcija sojeva Saccharomyces cerevisiae za produkciju aminokiselina nalik mikosporinu (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:565236

Jurić, Vanja. "Konstrukcija sojeva Saccharomyces cerevisiae za produkciju aminokiselina nalik mikosporinu." Master's thesis, University of Zagreb, Faculty of Science, 2019. https://urn.nsk.hr/urn:nbn:hr:217:565236

Jurić, Vanja. "Konstrukcija sojeva Saccharomyces cerevisiae za produkciju aminokiselina nalik mikosporinu." Master's thesis, University of Zagreb, Faculty of Science, 2019. https://urn.nsk.hr/urn:nbn:hr:217:565236

Jurić, V. (2019). 'Konstrukcija sojeva Saccharomyces cerevisiae za produkciju aminokiselina nalik mikosporinu', Master's thesis, University of Zagreb, Faculty of Science, accessed 21 May 2024, https://urn.nsk.hr/urn:nbn:hr:217:565236

Jurić V. Konstrukcija sojeva Saccharomyces cerevisiae za produkciju aminokiselina nalik mikosporinu [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2019 [cited 2024 May 21] Available at: https://urn.nsk.hr/urn:nbn:hr:217:565236

V. Jurić, "Konstrukcija sojeva Saccharomyces cerevisiae za produkciju aminokiselina nalik mikosporinu", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2019. Available at: https://urn.nsk.hr/urn:nbn:hr:217:565236

Metadata
TitleKonstrukcija sojeva Saccharomyces cerevisiae za produkciju aminokiselina nalik mikosporinu
Title (english)Construction of Saccharomyces cerevisiae strains for mycosporine-like amino acids production
AuthorVanja Jurić
MentorIrina Borodina (mentor)
MentorIvana Ivančić Baće (sumentor)
Committee memberIvana Ivančić Baće (predsjednik povjerenstva)
Committee memberSunčica Bosak (član povjerenstva)
Committee memberTomislav Ivanković (član povjerenstva)
Committee memberDuje Lisičić (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Biology)
Zagreb
Defense date and country2019-11-15, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Abstract
Aminokiseline nalik mikosporinu su spojevi poznati po upijanju UV zračenja te su prisutni u brojnim morskim mikroorganizmima koji luče ove sekundarne metabolite da bi se zaštitili u uvjetima stresa izazvanog UV zračenjem. Ovi spojevi tipično apsorbiraju na valnim duljinama od 310 nm do 360 nm. Njihova fotoprotektivna svojstva čine ih obećavajućim kandidatima za farmaceutske i kozmetičke aplikacije. Cilj ovog istraživanja bila je konstrukcija sojeva kvasca Saccharomyces cerevisiae koji će u kromosomima 10 i 11 imati stabilno integrirani heterologni biosintetski put iz cijanobakterije Anabaena variabilis za proizvodnju aminokiselina nalik mikosporinu. Konstruirana su tri integrativna plazmida koji sadržavaju gene za biosintetski put iz A. variabilis uz odgovarajuće konstitutivne promotore i terminatore. Put je ugrađen u haploidne i diploidne sojeve S. cerevisiae te u derivate tih sojeva koji eksprimiraju heterologni put za konzumaciju ksiloze kako bi uz glukozu mogli rasti i na mediju s ksilozom. Ispravnost plazmidnih konstrukata je potvrđena metodom analize kolonija PCR-om te sekvenciranjem. Metodom analize kolonija PCR-om je potvrđena ugradnja vektora u kromosome S. cerevisiae, a metodom tekućinske kromatografije visoke djelotvornosti (HPLC) te masenom spektrometrijom (LC-MS) detektirani su proizvedeni spojevi iz puta sinteze aminokiselina nalik mikosporinu u konstruiranim sojevima.
Abstract (english)
Mycosporine-like amino acids are compounds known for their UV-absorbing properties and are found in numerous marine microorganisms which produce these secondary metabolites as a way of dealing with UV-induced stress. Their absorption maxima typically range from 310 to 360 nm. The aim of this thesis was to construct Saccharomyces cerevisiae strains with heterologous biosynthetic mycosporine-like amino acids pathway from cyanobacterium Anabaena variabilis stably integrated into chromosomes 10 and 11. Three plasmids containing the pathway from A. variabilis with corresponding constitutive promoters and terminators were constructed. The pathway was integrated into haploid and diploid S. cerevisiae strains and into their derivatives which express a heterologous pathway for xylose consumption so that the strains would be able to grow on medium supplemented with xylose, in addition to glucose. Plasmid constructs were verified by colony PCR and sequencing. Colony PCR was used to confirm vector integration into S. cerevisiae chromosomes and high performance liquid chromatography (HPLC) combined with mass spectrometry (LC-MS) was used to detect the produced compounds from the mycosporine-like amino acids pathway in constructed yeast strains.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:565236
Study programmeTitle: Molecular Biology
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Embargo expiration date: 2021-11-15
Terms of useLicence In copyright icon
Created on2020-06-09 11:12:04