20.500.12556/DKUM-94520 Uporaba encima invertaza za razgradnjo saharoze v pretočnem reaktorju diplomsko delo Application of the enzyme invertase for the degradation of sucrose in a flow reactor V okviru diplomskega dela smo raziskovali vpliv pretoka, pH in temperature na presnovo saharoze z encimom invertaza. Cilj raziskovanja je bila optimizacija eksperimentalnih pogojev za čim večjo učinkovitost encimske razgradnje saharoze v pretočnem reaktorju. Saharozo smo razgrajevali z encimom invertaza, ki smo ga imobilizirali na silika gelu. Uspešnost imobilizacije smo določili z Bradfordovo metodo, pri kateri smo sprektrofotometrično določili količino encima, ki se je vezal na silika gel. Reakcije smo izvajali pri različnih temperaturah, pH-jih in pretokih s konstantno koncentracijo substrata (saharoze) v pretočnem mikroreaktorju pri čemer smo temperaturo zagotavljali z vodno kopeljo. Koncentracijo smo določili na podlagi rezultatov iz HPLC analize in jih vnesli v računalniški program. Podatek o presnovi smo ocenili iz koncentracije nastale fruktoze, ki je eden izmed produktov razpada saharoze. Tako smo potrdili hipotezo, da lahko z encimom invertaza uspešno razgrajujemo saharozo v pretočnem reaktorju. Dobili smo polinom, ki je predstavljal izhodno koncentracijo fruktoze kot funkcijo treh spremenljivk: pH, temperature in pretoka. Polinom predstavlja model, ki ga uporabljamo za napoved rezultatov v definiranih območjih veljavnosti. Prav tako nam je uspelo dokazati tudi dodatno hipotezo in sicer, da lahko z metodo odzivnih površin (centralno kompozitna metoda) zadovoljivo, natančno ter z izbranimi parametri opišemo model, ki opisuje nastanek fruktoze kot rezultat razgradnje saharoze z uporabo encima invertaze. Model lahko uporabimo tudi za optimizacijo procesa in izračunamo optimalne pogoje, pri katerih bi dosegli najvišjo presnovo oz. najvišjo koncentracijo fruktoze. Iz rezultatov je razvidno, da ima največji vpliv na koncentracijo fruktoze pH in da lahko dosežemo optimalno presnovo pri pH 4,76, temperaturi 40,39 °C in pretoku 1,20 mL/min. As part of this thesis, we investigated the influence of flow rate, pH and temperature on the degradation of sucrose using the enzyme invertase. Our aim was to optimize experimental conditions for maximizing the efficiency of enzymatic sucrose degradation in a flow reactor. Sucrose was hydrolyzed using invertase immobilized on silica gel. Efficiency of the immobilization was determined by the Bradford method, where we spectrophotometrically measured the amount of enzyme bound to the silica gel. Reactions were carried out at different temperatures, pH values and flow rates, with a constant substrate (sucrose) concentration in the flow microreactor, where the temperature was regulated with a water bath. The concentration was determined with HPLC analysis and processed using computational software. The extent od sucrose conversion was evaluated from the concentration of fructose, which is one of the products of sucrose degradation. This confirmed the hypothesis that sucrose can be successfully hydrolyzed with invertase in a flow reactor. A polynomial equation was obtained, representing the degradation of sucrose as a function of three variables: pH, temperature, and flow rate. The polynomial serves as a model that can be used to predict results with the defined range of validity. We were also able to confirm the additional hypothesis, which states that with the response surface method (central composite design) we can satisfactorily and accuratelly describe the model for fructose formation as a result of sucrose degradation by invertase. The model can also be used for process optimization, where we can calculate the optimal conditions for achieving the highest conversion or the highest fructose concentration. The results showed that pH has the strongest effect on fructose concentration, and optimal conversion can be achieved at pH 4,76, temperature 40,39 °C, and flow rate of 1,20 mL/min. invertaza saharoza presnova fruktoza metoda odzivnih površin invertase sucrose degradation fructose response surface method true true false [A. Škrinjar] Slovenski jezik Angleški jezik Diplomsko delo/naloga 2025-08-20 11:20:30 2025-09-18 11:47:20 2025-09-30 03:33:28 0000-00-00 00:00:00 2025 0 Maribor 0 Maribor 1 spletni vir (1 datoteka PDF (VIII, 28 f.)) 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 66.098:[577.124:577.15](043.2) 250875907 UN_Skrinjar_Amadeja_2025.pdf UN_Skrinjar_Amadeja_2025.pdf 1 BDEF7A168911A0C64BF703C2B3047A38 6835391e6afea89e68d210798b2597e916157a9ebf8a2883f40a182f8ce12cd9 c5758efd-93ec-11f0-821d-00155d000105 https://dk.um.si/Dokument.php?lang=slv&id=198707 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0