20.500.12556/DKUM-94320 Imobilizacija encima invertaza na zeolitni nosilec diplomsko delo visokošolskega strokovnega študijskega programa I. stopnje Immobilization of invertase enzyme on a zeolite carrier Encimi so zaradi svoje specifičnosti, učinkovitosti in sposobnosti delovanja pri blagih pogojih široko uporabljeni v različnih industrijskih procesih. Njihova uporaba v prosti obliki je pogosto omejena zaradi nizke stabilnosti ter oteženega ločevanja iz reakcijskega medija. V okviru diplomskega dela smo preučevali imobilizacijo encima invertaze na zeolitni nosilec ZP-4A, ki se zaradi visoke specifične površine, toplotne obstojnosti in možnosti kemijske funkcionalizacije izkazuje kot obetaven material za biokatalitske aplikacije. Encim smo s kovalentno vezavo pritrdili na nosilec z uporabo (3-aminopropil)trietoksisilana kot silanizacijskega reagenta in glutaraldehida kot zamreževalnega sredstva. Učinkovitost imobiliziranega encima smo preverili na reakciji hidrolize saharoze pri različnih vrednostih pH in temperature, s ciljem oceniti vpliv teh parametrov na katalitsko aktivnost. Koncentracije produktov smo določili z uporabo tekočinske kromatografije visoke ločljivosti (HPLC), spremembe strukture in površine nosilca pa z metodo infrardeče spektroskopije s Fourierjevo transformacijo (FTIR), Brunauer–Emmett–Tellerjevo fizisorpcijo dušika (BET), vrstično elektronsko mikroskopijo (SEM) ter elementno analizo (EA). Iz rezultatov je razvidno, da je bila imobilizacija uspešna in da je imobilizirana invertaza ohranila učinkovitost v širšem območju eksperimentalnih pogojev. Največja aktivnost je bila dosežena pri pH = 4,5 in temperaturi 30 °C. Rezultati encimskih reakcij ter analiz površine nosilca (SEM, BET, EA) potrjujejo, da je bil zeolit ZP-4A ustrezen za imobilizacijo, saj je encim na njem ohranil stabilnost in omogočil večkratno uporabo. Enzymes are widely used in various industrial processes due to their specificity, efficiency, and ability to function under mild conditions. However, their application in free form is often limited by low stability and the difficulty of separation from the reaction medium. In this thesis, we investigated the immobilization of the enzyme invertase onto the zeolite support ZP-4A, which, owing to its high specific surface area, thermal stability, and potential for chemical functionalization, represents a promising material for biocatalytic applications. The enzyme was covalently attached to the support using (3-aminopropyl)triethoxysilane as the silanization reagent and glutaraldehyde as the crosslinking agent. The performance of the immobilized enzyme was evaluated in the hydrolysis of sucrose under different pH and temperature conditions, with the aim of assessing the influence of these parameters on catalytic activity. Product concentrations were determined by high-performance liquid chromatography (HPLC), while changes in the structure and surface of the support were characterized by Fourier-transform infrared spectroscopy (FTIR), Brunauer–Emmett–Teller (BET) nitrogen physisorption, scanning electron microscopy (SEM), and elemental analysis (EA). The results showed that immobilization was successful and that immobilized invertase retained its catalytic efficiency over a wider range of experimental conditions. The highest activity was observed at pH = 4,5 and 30 °C. The results of enzymatic reactions and support characterization (SEM, BET, EA) confirmed that zeolite ZP-4A is a suitable carrier for immobilization, as the enzyme maintained stability on its surface and enabled repeated use. invertaza imobilizacija encima zeolit ZP-4A saharoza invertase enzyme immobilization zeolite ZP-4A sucrose true true false [V. Rihtarič] Slovenski jezik Angleški jezik Diplomsko delo/naloga 2025-08-13 13:31:32 2025-09-05 14:51:31 2025-09-10 03:36:35 0000-00-00 00:00:00 2025 0 Maribor 0 Maribor 1 spletni vir (1 datoteka PDF (VIII, 38 str.)) 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 577.15(043.2) 248240643 VS_Rihtaric_Viktorija_2025.pdf VS_Rihtaric_Viktorija_2025.pdf 1 B4E8946F43FF979964B7BEDEBA5EAEFB 5d7f5194f589bf1b92ae02b852f714a33b4eee7c49bcc7b1c67f30af94c0b737 87a48687-89b1-11f0-8088-00155d000105 https://dk.um.si/Dokument.php?lang=slv&id=197661 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0