Polimerizacija flavonoida katalizirana enzimom peroksidaza porijeklom iz hrena

undergraduate thesis

• Title (english): Polymerization of flavonoids catalysed by horseradish peroxidase
• Author: Ana Boltek
• Mentor: Anita Šalić (mentor)
• Committee member: Anita Šalić (predsjednik povjerenstva)
• Committee member: Andreja Žužić (član povjerenstva)
• Committee member: Ivan Karlo Cingesar (član povjerenstva)
• Granter: University of Zagreb; Faculty of Chemical Engineering and Technology; Zagreb
• Defense date and country: 2024-09-23, Croatia
• Scientific / art field, discipline and subdiscipline: TECHNICAL SCIENCES; Chemical Engineering

Abstract

Flavoni, u koje se ubraja i apigenin, prepoznati su kao obećavajući kandidati za prevenciju metastaza raka zahvaljujući njihovoj sposobnosti inhibicije ključnih signalnih puteva. Ipak, prirodni oblik apigenina pokazuje slabu topljivost i stabilnost, što ograničava njegovu terapijsku učinkovitost. Kako bi se prevladala ta ograničenja, apigenin se može kemijski modificirati u biflavonoide i derivate poliapigenina pomoću enzima poput lakaze i peroksidaze porijeklom iz hrena (HRP). Cilj ovog rada bio je provesti polimerizaciju apigenina pomoću enzima peroksidaza porijeklom iz hrena. Polimerizacija je provedena u kotlastom reaktoru i mikroreaktoru, a tijekom procesa praćene su karakteristične veličine procesa kao što su konverzija, reakcijsko vrijeme/vrijeme zadržavanja i procesna stabilnost enzima. Oligomerna struktura flavonoida potvrđena je infracrvenom spektroskopijom s Fourierovom transformacijom (FT-IR) i kapljevinskom kromatografijom visoke djelotvornosti (HPLC-DAD). U drugom dijelu istraživanja ispitana je reakcijska kinetika polimerizacije apigenina metodom početnih brzina te su procijenjeni kinetički parametri. Kinetika polimerizacije apigenina katalizirana enzimom peroksidaza porijeklom iz hrena opisana je Michaelis–Menteničinom kinetikom. Praćenjem procesne stabilnosti enzima, primijećen je pad aktivnosti te je procijenjena konstanta deaktivacije enzima peroksidaza porijeklom iz hrena. Za reakciju provedenu u kotlastom reaktoru i mikroreaktoru postavljen je odgovarajući reaktorski model koji se sastojao od kinetičkog modela, deaktivacije enzima i bilanci tvari. Ocjena valjanosti modela provedena je na dva seta nezavisnih eksperimenata.

Abstract (english)

Flavones, which include apigenin, are recognized as promising candidates for the prevention of cancer metastasis due to their ability to inhibit key signaling pathways. However,
the natural form of apigenin shows poor solubility and stability, which limits its therapeutic efficacy. To overcome these limitations, apigenin can be chemically modified into biflavonoids and polyapigenin derivatives using enzymes such as laccase and horseradish-derived peroxidase enzyme (HRP). The aim of this work was to polymerize apigenin using horseradish-derived peroxidase enzymes. The polymerization was carried out in a batch reactor and a microreactor, and during the process characteristic parameters such as conversion, reaction time/retention time and process stability of the enzyme were monitored. Oligomeric structure of flavonoids was confirmed by Fourier transform infrared spectroscopy (FT-IR) and high performance liquid chromatography (HPLC-DAD). In the second part of the study, the reaction kinetics of apigenin polymerization was investigated using the initial rate method and the kinetic parameters were estimated. The kinetics of apigenin polymerization catalyzed by the enzyme horseradish peroxidase was described by Michaelis–Menten kinetics. When monitoring the process stability of the enzyme, a decrease in activity was observed and the deactivation constant of the horseradish peroxidase enzyme was estimated. A suitable reactor model was proposed for the reaction carried out in a batch reactor and a microreactor, which consisted of a kinetic model, enzyme deactivation and a mass balance. The validation of the developed model was performed using two independent experiments.

• Keywords: flavoni; apigenin; enzim peroksidaza; polimerizacija
• Keywords (english): flavones; apigenin; peroxidase enzyme; polymerization
• Language: croatian
• URN:NBN: urn:nbn:hr:149:302147
• Project: Number: UNIN-BIOTEH-24-1-1; Title: Biotransformacije te biološka aktivnost metiliranih flavona i njihovih dimera- druga faza istraživanja; Title: Biotransformations and biological activity of methylated flavones and their dimers-2nt phase; Leader: Dunja Šamec; Jurisdiction: Croatia; Funder: Sveučilište Sjever
• Study programme: Title: Environmental Engineering - Undergraduate study; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka ekoinženjerstva (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka ekoinženjerstva)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-11-17 19:11:54