Izolacija kvasovk Saccharomyces cerevisiae iz neobičajnih naravnih habitatov

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Ključne besede

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Jezik:	Slovenski jezik
Leto izida:	2012
Izvor:	Ljubljana
Tipologija:	2.11 - Diplomsko delo
Organizacija:	UL PEF - Pedagoška fakulteta
Založnik:	[B. Finžgar]
UDK:	57:582.28(9)(043.2)
COBISS:	9146697
Št. ogledov:	1977
Št. prenosov:	387
Ocena:	0 (0 glasov)
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Sekundarni jezik:	Angleški jezik
Sekundarni naslov:	Isolation of yeast Saccharomyces cerevisiae from unusual natural habitats
Sekundarni povzetek:	Baker yeast Saccharomyces cerevisiae has been an eukarontic experimental organism since 1960s, becoming even more significant with the determination of its complete nucleotide genome sequence in 1996. Even though its biochemical function in the fermentation process had long remained unclear, its metabolism and products (eg. bread, beer, wine) have been used for millennia. S. cerevisiae yeast represents an important organism for production of recombinant proteins (gene manipulation). Moreover, it exhibits great potential for biofuels and the process of bioethanol production from waste materials such as lignocellulose. The yeast is generally found in anthropogenic environments, yet little is known about its ecology. The purpose of this given thesis was to perform a selective isolation of the S. cerevisiae species from both anthropogenic and less common natural habitats. The aim was to aquire strains with metabolic properties that would be potentially uncommon for the species, yet effective in bioethanol synthesis. In the scope of the research, 273 collected samples were divided into 27 different groups. Through the use of a selective growth medium with the addition of 10 % ethanol, 122 yeast cultures were isolated and initially defined according to their morphology (colour, shape) and micromorphology (cell size and shape). In order to identify the strains according to their homothallic or heterothallic properties, the presence of sexual structures (asci, ascospores) was examined on acetate agar. The concluding identification took place at the molecular genetic level through the comparison of nucleotide sequencing of the regions of internal transcribed spacers 1 and 2, including the 5.8S rDNA sequence (ITS rDNA). From the 122 samples, 44 strains of the S. cerevisiae yeast were isolated. The S. cerevisiae species was typically isolated from vinicultural samples and industrial environment waters, whereas individual strains were isolated also from dairy products, freshly squeezed non-pasteurized juices, the ground beneath fruit trees, fresh or rotten fruits and vegetables, insects, vinegar, decomposed wood, and from silage. The S. cerevisiae strains were determined according to their assimilation, fermentation and temperature profiles, and the differences among individual strains were examined.
Sekundarne ključne besede:	biology;biologija;
Vrsta datoteke:	application/pdf
Vrsta dela (COBISS):	Diplomsko delo
Komentar na gradivo:	Univ. Ljubljana, Pedagoška fak., Biotehniška fak., Biologija in gospodinjstvo
Strani:	XII, 79 f., [7] f. pril.
Vrsta dela (ePrints):	thesis
Naslov (ePrints):	Isolation of yeast Saccharomyces cerevisiae from unusual natural habitats
Ključne besede (ePrints):	glive
Ključne besede (ePrints, sekundarni jezik):	fungi
Povzetek (ePrints):	Pivska kvasovka Saccharomyces cerevisiae je že od 60. let 20 stoletja evkariontski eksperimentalni organizem, ki je postal leta 1996 z določitvijo celotnega nukleotidnega zaporedja genoma še pomembnejši. Čeprav njena biokemijska vloga v fermentacijskem procesu dolgo časa ni bila pojasnjena, smo njen metabolizem in produkte uporabljali že tisočletja (npr. kruh, pivo, vino). Kvasovka S. cerevisiae je pomembno orodje za pridobivanje rekombinantnih proteinov (genska manipulacija). Predstavlja pa tudi velik potencial na področju biogoriv, proizvodnje bioetanola iz odpadnih materialov, kot je lignoceluloza. Kvasovka sicer pogosto naseljuje antropogena okolja, manj pa je znanega o njeni ekologiji. Namen diplomskega dela je bila selektivna izolacija vrste S. cerevisiae iz antropogenih in manj običajnih naravnih okolij. Cilj naloge je bila pridobitev sevov s potencialno neobičajnimi metaboličnimi lastnostmi za to vrsto, ki bi bile uporabne pri sintezi bioetanola. V sklopu diplomske naloge smo nabrali 273 različnih vzorcev, ki smo jih razdelili v 27 različnih skupin. Z uporabo selektivnega gojišča z dodatkom 10 % etanola smo izolirali 122 kvasnih kultur, ki smo jih sprva proučevali na ravni morfologije (barva, oblika) in mikromorfologije kolonij (velikost in oblika celic). Preverjali smo, ali so sevi homo- ali heterotalični ter v ta namen proučevali prisotnost spolnih struktur (aski, askospore) na acetatnem agarju. Dokončna identifikacija je potekala na molekularno-genetskem nivoju s primerjavo nukleotidnih zaporedij regij notranjih distančnikov 1 in 2, vključno s 5,8S rDNA zaporedjem (ITS rDNA). Iz 122 vzorcev smo izolirali 44 sevov kvasovke vrste S. cerevisiae. Vrsto S. cerevisiae smo najpogosteje izolirali iz vinogradniških vzorcev ter vod industrijskih okolij, posamezne seve pa smo izolirali tudi iz mlečnih izdelkov, sveže stisnjenih nepasteriziranih sadnih sokov, tal pod sadnim drevjem, sadjem in zelenjavo, iz žuželk, domačega kisa, razgrajenega lesa, svežega in gnilega sadja in zelenjave ter iz silaže. Sevom S. cerevisiae smo določili asimilacijski, fermentacijski in temperaturni profil ter opazovali odstopanja med posameznimi sevi.
Povzetek (ePrints, sekundarni jezik):	Baker yeast Saccharomyces cerevisiae has been an eukarontic experimental organism since 1960s, becoming even more significant with the determination of its complete nucleotide genome sequence in 1996. Even though its biochemical function in the fermentation process had long remained unclear, its metabolism and products (eg. bread, beer, wine) have been used for millennia. S. cerevisiae yeast represents an important organism for production of recombinant proteins (gene manipulation). Moreover, it exhibits great potential for biofuels and the process of bioethanol production from waste materials such as lignocellulose. The yeast is generally found in anthropogenic environments, yet little is known about its ecology. The purpose of this given thesis was to perform a selective isolation of the S. cerevisiae species from both anthropogenic and less common natural habitats. The aim was to aquire strains with metabolic properties that would be potentially uncommon for the species, yet effective in bioethanol synthesis. In the scope of the research, 273 collected samples were divided into 27 different groups. Through the use of a selective growth medium with the addition of 10 % ethanol, 122 yeast cultures were isolated and initially defined according to their morphology (colour, shape) and micromorphology (cell size and shape). In order to identify the strains according to their homothallic or heterothallic properties, the presence of sexual structures (asci, ascospores) was examined on acetate agar. The concluding identification took place at the molecular genetic level through the comparison of nucleotide sequencing of the regions of internal transcribed spacers 1 and 2, including the 5.8S rDNA sequence (ITS rDNA). From the 122 samples, 44 strains of the S. cerevisiae yeast were isolated. The S. cerevisiae species was typically isolated from vinicultural samples and industrial environment waters, whereas individual strains were isolated also from dairy products, freshly squeezed non-pasteurized juices, the ground beneath fruit trees, fresh or rotten fruits and vegetables, insects, vinegar, decomposed wood, and from silage. The S. cerevisiae strains were determined according to their assimilation, fermentation and temperature profiles, and the differences among individual strains were examined.
Ključne besede (ePrints, sekundarni jezik):	fungi
ID:	8308930