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Izražanje genov za biosintezo likopena v kvasovki Yarrowia lipolytica
Prendl, Patricija (Avtor), Petrovič, Uroš (Mentor) Več o mentorju... Povezava se odpre v novem oknu, Magdevska, Vasilka (Komentor)

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Izvleček
Terpeni so velika skupina organskih spojin, ki jih proizvajajo različni mikroorganizmi, rastline in tudi nekatere žuželke. Biosinteza poteka preko molekule izopentenil difosfat (IPP), ki je glavni gradnik za vse vrste terpenov. Terpeni se uporabljajo kot prehranska dopolnila v prehrani ljudi in živali ter v farmacevtski in kozmetični industriji. Med terpene uvrščamo tudi karotenoide, kamor spada likopen. Likopen je rdeče barvilo, ki ima izmed vseh karotenoidov najmočnejše antioksidativno delovanje. Z biotehnološkega vidika je za produkcijo likopena zanimiva oleogena kvasovka Yarrowia lipolytica, ki je sposobna v lipidnih kapljicah kopiči lipide do 20 % svoje suhe teže. Y. lipolytica po naravni metabolni poti ne producira likopena, saj se potencialna biosintezna pot likopena ustavi na stopnji intermediata geranilgeranil difosfata. Za preusmeritev metabolne poti do sinteze likopena so potrebni geni crtE, crtB in crtI. Ti geni imajo zapis za encime, ki katalizirajo metabolno pot od geranilgeranil difosfata do likopena. Gen crtE ima zapis za encim geranilgeranil difosfat sintazo, crtB fitoen sintazo in crtI fitoen desaturazo. V okviru magistrske naloge smo z metodo »Golden Gate« sestavili šest plazmidov, izmed katerih je vsak vseboval heterologne gene crtB, crtE in crtI, ki smo jih s transformacijo vstavili v Y. lipolytica. Uspeli smo pridobiti transformante, ki so sintetizirale likopen. V dušik omejujočem gojišču je bila produkcija likopena večja kot v gojišču YEPD in v gojišču z glicerolom kot edinim virom ogljika.

Jezik:Slovenski jezik
Ključne besede:kvasovke, Yarrowia lipolytica, metabolni inženiring, metoda »Golden Gate«, produkcija likopena
Vrsta gradiva:Magistrsko delo/naloga (mb22)
Organizacija:BF - Biotehniška fakulteta
Leto izida:2019
COBISS.SI-ID:4987983 Povezava se odpre v novem oknu
Število ogledov:310
Število prenosov:222
Metapodatki:XML RDF-CHPDL DC-XML DC-RDF
 
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Sekundarni jezik

Jezik:Angleški jezik
Naslov:Expression of lycopene biosynthesis genes in yeast Yarrowia lipolytica
Izvleček:
Terpenes are a large group of organic compounds produced by various microorganisms, plants and also some insects. Their biosynthesis is performed through the isopentenyl diphosphate (IPP) molecule, which is the main precursor for all terpenes. Terpenes are used as food supplements for humans and animals and in the pharmaceutical and cosmetic industry. Among terpenes are also carotenoids, which include lycopene. Lycopene is a red dye that has the highest antioxidant activity among all carotenoids. The yeast Yarrowia lipolytica is an interesting host for lycopene production as is capable of accumulating lipids in lipid droplets up to 20 % of its dry cell weight. Y. lipolytica does not produce lycopene naturally, as the potential biosynthetic path stops at the level of intermediate geranylgeranyl diphosphate. To divert metabolic pathway to synthesize lycopene genes crtE, crtB and crtI are required. These genes are coding enymes that catalyze production lycopene from geranylgeranyl diphosphate. In this study we assembled six plasmid constructs with the Golden Gate technique. Plasmids containing heterologous genes crtB, crtE and crtI were transformed into Y. lipolytica. We successfully screened transformants that synthesized lycopene and compared the production of lycopene in various production media. In the nitrogen-limited medium the production of lycopene was higher than in YEPD medium and in the medium with gylcerol as the carbon source.

Ključne besede:yeast, Yarrowia lipolytica, metabolic engineering, Golden Gate Assembly, lycopene production

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