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Title:Uporaba encima invertaza za razgradnjo saharoze v pretočnem reaktorju : diplomsko delo
Authors:ID Škrinjar, Amadeja (Author)
ID Nemet, Andreja (Mentor) More about this mentor... New window
ID Pečar, Darja (Comentor)
Files:.pdf UN_Skrinjar_Amadeja_2025.pdf (2,03 MB)
MD5: BDEF7A168911A0C64BF703C2B3047A38
 
Language:Slovenian
Work type:Bachelor thesis/paper
Typology:2.11 - Undergraduate Thesis
Organization:FKKT - Faculty of Chemistry and Chemical Engineering
Abstract: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.
Keywords:invertaza, saharoza, presnova, fruktoza, metoda odzivnih površin
Place of publishing:Maribor
Place of performance:Maribor
Publisher:[A. Škrinjar]
Year of publishing:2025
Number of pages:1 spletni vir (1 datoteka PDF (VIII, 28 f.))
PID:20.500.12556/DKUM-94520 New window
UDC:66.098:[577.124:577.15](043.2)
COBISS.SI-ID:250875907 New window
Publication date in DKUM:18.09.2025
Views:147
Downloads:25
Metadata:XML DC-XML DC-RDF
Categories:KTFMB - FKKT
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Licences

License:CC BY-NC-ND 4.0, Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
Link:http://creativecommons.org/licenses/by-nc-nd/4.0/
Description:The most restrictive Creative Commons license. This only allows people to download and share the work for no commercial gain and for no other purposes.
Licensing start date:29.08.2025

Secondary language

Language:English
Title:Application of the enzyme invertase for the degradation of sucrose in a flow reactor
Abstract: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.
Keywords:invertase, sucrose, degradation, fructose, response surface method


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