magistrsko delo
Melani Dolgov (Author), Maja Leitgeb (Mentor), Mateja Primožič (Co-mentor), Pia Keršič (Co-mentor)

Abstract

Sekundarni metaboliti so naravne spojine, ki jih sintetizirajo mikroorganizmi, vključno z bakterijami, glivami, kvasovkami in rastlinami. Čeprav ti produkti niso nujno potrebni za osnovne biološke funkcije, kot so sinteza lipidov, aminokislin, ogljikovih hidratov in nukleinskih kislin, imajo pomembno vlogo v različnih industrijskih aplikacijah. Kvasovka Saccharomyces cerevisiae je ključni mikroorganizem v biotehnologiji zaradi svoje sposobnosti pretvorbe sladkorjev v etanol in ogljikov dioksid ter njene prilagodljivosti na različne okoljske pogoje. Cilj magistrskega dela je bil optimizirati proizvodnjo sekundarnih metabolitov s pomočjo S. cerevisiae v šaržnem bioreaktorju. Raziskava je obsegala preučevanje vpliva različnih vrst mešal, začetnih koncentracij inokuluma ter hitrosti mešanja na rast celične mase in proizvodnjo sekundarnih metabolitov. Med optimizacijo smo spremljali spremembe pH med fermentacijo ter izvajali UV-VIS spektrofotometrične analize za pridobitev rezultatov o količini celokupnih proteinov, antioksidativnem potencialu in aktivnosti encima alkohol dehidrogenaze (ADH). Antioksidativni potencial proizvedenih sekundarnih metabolitov smo določili z DPPH metodo , za določanje koncentracije celokupnih proteinov je bila uporabljena Bradfordova metoda, aktivnost znotrajceličnega encima ADH pa je bila določena s pomočjo encimskega testa. Optimalni parametri za doseganje najboljših rezultatov vključujejo hitrost mešanja 200 rpm, uporabo dvojnega mešala in začetno koncentracijo inokuluma 6,62 ∙ 107 CFU/mL. Ti parametri so bili potrjeni v zaključnem eksperimentu, kjer smo ponovno gojili S. cerevisiae pod enakimi pogoji, kar je potrdilo ponovljivost gojenja in učinkovitost izbranih parametrov. Prav tako so rezultati analitskih metod pokazali, da so permeabilizirani vzorci dosegli najvišjo antioksidativno aktivnost, najvišjo koncentracijo proteinov in največjo aktivnost ADH. Na podlagi teh ugotovitev smo zaključili, da višja hitrost mešanja, uporaba dvojnega mešala in večja začetna koncentracija inokuluma zagotavljajo optimalne rezultate v proizvodnji sekundarnih metabolitov.

Keywords

bioreaktor;procesni parametri;optimizacija;S. cerevisiae;alkohol dehidrogenaza;sekundarni metaboliti;magistrske naloge;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UM FKKT - Faculty of Chemistry and Chemical Engineering
Publisher: [M. Dolgov]
UDC: [601.2:577.121]:602.42-7(043.2)
COBISS: 224391171 Link will open in a new window
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Other data

Secondary language: English
Secondary title: Optimization of process parametres for production of secondary metabolites in a batch bioreactor
Secondary abstract: Secondary metabolites are natural compounds synthesized by microorganisms, including bacteria, fungi, yeast, and plants. Although these products are not necessarily essential for fundamental biological functions such as the synthesis of lipids, amino acids, carbohydrates, and nucleic acids, they play a significant role in various industrial applications. The yeast Saccharomyces cerevisiae is a key microorganism in biotechnology due to its ability to convert sugars into ethanol and carbon dioxide, as well as its adaptability to various environmental conditions. The aim of this thesis was to optimize the production of secondary metabolites when cultivating S. cerevisiae in a batch bioreactor. The study involved examining the impact of different types of agitators, initial inoculum concentrations, and agitation speeds on cell biomass growth and secondary metabolite production. During optimization, pH changes during fermentation were monitored, and UV-VIS spectrophotometric analyses were performed to obtain results for the amount of total proteins, for the antioxidative potential and for the enzyme activity of the enzyme alcohol dehydrogenase (ADH). To determine the antioxidative potential of the produced secondary metabolites, the DPPH method was used, total protein concentration was measured with the Bradford method, and an enzymatic assay was conducted to measure ADH activity. The optimal parameters for achieving the best results include an agitation speed of 200 rpm, the use of a dual agitator, and an initial inoculum concentration of 6.62 ∙ 107 CFU/mL. These parameters were confirmed in a final experiment, where S. cerevisiae was grown under the same conditions, validating the reproducibility and effectiveness of the chosen parameters. Additionally, the results of the analytical methods demonstrated that permeabilized samples achieved the highest antioxidant activity, highest protein concentration, and highest ADH activity. Based on these findings, it was concluded that higher agitation speed, the use of a dual agitator, and higher initial inoculum concentration ensure optimal results in secondary metabolite production.
Secondary keywords: bioreaktor;procesni parametri;optimizacija;S. cerevisiae;alkohol dehidrogenaza;sekundarni metaboliti;
Type (COBISS): Master's thesis/paper
Thesis comment: Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Pages: 1 spletni vir (1 datoteka PDF (X, 61 str.))
ID: 25033884