20.500.12556/RUL-162297 Primerjalna analiza proteoma prostih in kapsuliranih kvasovk Saccharomyces cerevisiae med alkoholno fermentacijo Comparative proteomic analysis of free and encapsulated yeasts Saccharomyces cerevisiae during alcoholic fermentation Kvasovke S. cerevisiae so med alkoholno fermentacijo izpostavljene različnim stresnim dejavnikom, kot so visoka temperatura ter visoka koncentracija substratov in nastalih produktov. Pred vsem tem lahko kvasovke učinkovito zaščitimo z mikrokapsulacijo, ki kot tehnika pridobiva vse večjo vlogo v biotehnologiji in živilstvu. V magistrski nalogi smo z napravo z elektrostatično vibracijsko šobo kapsulirali kvasovke S. cerevisiae v alginat, nekaterim vzorcem pa smo dodali tudi različne ciklodekstrine. Spremljali smo potek alkoholne fermentacije in vpliv nizke (8 %) in visoke (28 %) koncentracije glukoze v gojišču ter dodatek β-CD, β-CD citrata in β-CD maleata na izkoristek alkoholne fermentacije. Sproščanje kvasovk iz mikrokroglic smo spremljali z merjenjem OD650, količino nastalega etanola in porabljene glukoze pa smo analizirali z metodo HPLC. Z uporabo metode LC-MS/MS smo vse omenjene vplive analizirali še na proteomski ravni. Ugotovili smo, da kapsulacija z alginatom uspešno zaščiti kvasovke pred visoko koncentracijo glukoze in omogoča doseganje višjih izkoristkov fermentacije (do 97 %). To so nakazale tudi povečane ravni proteinov, vključenih v biosintezo aminokislin in glikolizo/glukoneogenezo. Dodatek 0,2 % ciklodekstrinov vpliva na povečano prepustnost hidrogelov, kjer so kapsulirane kvasovke dosegale primerljivo rast, manjši izkoristek ter znižano ravnen skoraj 70 % proteinov v primerjavi s prostimi kvasovkami. Nadalje je smotrno preučiti vpliv večjih in manjših koncentracij dodanih ciklodekstrinov, več bioloških ponovitev med daljšim spremljanim obdobjem fermentacije in vpliv populacije kvasovk vzdolž premera kroglice. S. cerevisiae yeasts are exposed to various stress factors during alcoholic fermentation, such as high temperature and high concentration of substrates and products. Yeast can be effectively protected from all this with microencapsulation, which is gaining an increasing role in biotechnology and food industry. In the master's thesis, yeasts S. cerevisiae were encapsulated in alginate using a device with an electrostatic vibration nozzle, and various cyclodextrins were added to some samples. We monitored the course of alcoholic fermentation and the effect of low (8 %) and high (28 %) glucose concentration in the culture medium and the addition of β-CD, β-CD citrate, and β-CD maleate on the yield of alcoholic fermentation. The release of yeast from the microspheres was monitored by measuring the OD650 and the amount of ethanol produced, and glucose consumed was analysed by the HPLC method. Using the LC-MS/MS method, we analysed the effects of all the mentioned factors on the proteomic level. We found that encapsulation with alginate successfully protects yeasts from high glucose concentrations and enables higher fermentation yields (up to 97%). This was also indicated by increased levels of proteins involved in amino acid biosynthesis and glycolysis/gluconeogenesis. The addition of 0,2 % cyclodextrins affects the increased permeability of the hydrogels, where the encapsulated yeasts achieved comparable growth, a lower yield, and a reduced level of almost 70 % of proteins compared to free yeasts. Furthermore, it is expedient to study the influence of higher and lower concentrations of added cyclodextrins, more biological repetitions during a longer monitored period of fermentation and the influence of the yeast population along the bead diameter. kvasovke Saccharomyces cerevisiae mikrokapsulacija alginat ciklodekstrini alkoholna fermentacija visoka koncentracija glukoze etanol izkoristek raven proteinov yeast Saccharomyces cerevisiae microencapsulation alginate cyclodextrins alcoholic fermentation high glucose concentration ethanol yield protein level true false false Slovenski jezik Angleški jezik Magistrsko delo/naloga 2024-09-21 07:15:18 2024-09-21 07:15:32 2024-09-24 10:35:12 0000-00-00 00:00:00 2024 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 246051 208679683 43682.pdf 43682.pdf 1 FC7B0546A029E7544BFCF506125FE273 1d42b00fe8207ee05b0acf9cd9ee000093378b101157bf0182cc65bb6f367282 7afd0a97-77d8-11ef-b232-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=191503 Biotehniška fakulteta 0 0 0