20.500.12556/DKUM-90497 Imobilizacija l-asparaginaze na magnetne nanodelce magistrsko delo Immobilization of l-asparaginase on magnetic nanoparticles V okviru zaključnega dela smo izvedli optimizacijo imobilizacije l-asparaginaze (l-ASNaze) na aminoksilanske magnetne nanodelce (AMN-MNPs). Postopek imobilizacije sestoji iz dveh korakov, funkcionalizacije AMN-MNPs in imobilizacije encima. V prvem koraku smo funkcionalizirali AMN-MNPs, v drugem koraku smo nanje imobilizirali l-ASNazo. Najprej smo v teoretičnem delu opisali lastnosti l-ASNaze, prikazali njeno strukturo, mehanizem delovanja in uporabo v različnih panogah. V nadaljnje smo opisali imobilizacijo na magnetne nanodelce (MNP), pri čemer smo se osredotočili na AMN-MNPs. Podan je tudi pregled literature o imobilizaciji l-ASNaze na MNPs. Pri eksperimentalnem delu smo proučevali vpliv različnih parametrov, kot so koncentracija mrežnega povezovalca glutaraldehida (GA), vrtilna hitrost, čas in temperatura imobilizacije, z namenom doseganje čim višje aktivnosti imobiliziranega encima in učinkovitost imobilizacije. Zanimalo nas je kakšno aktivnost encima dosežemo ob dodatku ogrodnih proteinov, ki delujejo kot stabilizatorji. Uporabili smo goveji serumski albumin (BSA) in albumin iz jajčnih beljakov (EA). V nadaljnje smo proučili termično stabilnost proste in imobilizirane l-ASNaze pri različnih časih inkubacije in pri različnih temperaturah. Prosto in imobilizirano l-ASNazo smo vzpostavili na magnetno polje in primerjali aktivnosti l-ASNaze pred in po vzpostavitvi. As part of the final work, we performed optimization of the immobilization of the enzyme L-asparaginase (ASNase) on aminosilane magnetic nanoparticles (AMN-MNPs). The immobilization process consists of two steps, AMN-MNPs functionalization and enzyme immobilization. In the first step, AMN-MNPs were functionalized, and in the second step, ASNase was immobilized on them. First, in the theoretical part, we described the properties of ASNase, showed its structure, mechanism of function and use in various branches. We further described the immobilization on magnetic nanoparticles (MNPs), focusing on AMN-MNPs. A literature review on the immobilization of ASNase on MNPs is also given. In the experimental work, we studied the influence of various parameters, such as the concentration of the crosslinker glutaraldehyde (GA), rotation speed, time and temperature of immobilization, with the aim of achieving the highest possible activity of the immobilized enzyme and immobilization efficiency. We were interested in what kind of enzyme activity is achieved with the addition of framework proteins that act as stabilizers. Bovine serum albumin (BSA) and egg white albumin (EA) were used. We further studied the thermal stability of free and immobilized ASNase at different incubation times and at different temperatures. Free and immobilized ASNase were exposed to a magnetic field and ASNase activities were compared before and after exposure. The final work contains a discussion of the results and graphically displayed results. l-asparaginaza magnetni nanodelci stabilnost kovalentna vezava funkcionalizacija imobilizacija L-asparaginase magnetic nanoparticles stability covalent binding functionalization immobilization true true false [M. Verdev] Slovenski jezik Angleški jezik Magistrsko delo/naloga 2024-09-05 16:06:24 2024-10-02 12:08:19 2024-10-19 03:10:54 0000-00-00 00:00:00 2024 0 Maribor 0 Maribor 1 spletni vir (1 datoteka PDF (X, 41 f.)) 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 577.151:544.142.2(043.2) 212127235 MAG_Verdev_Maja_2024.pdf MAG_Verdev_Maja_2024.pdf 1 3855B1E9A914CCF09F1F7DE09DECF1E7 6827230a0a7fde138466487729a00901380bdc66784adda243b602fc3933b984 e3d7acf5-8012-11ef-a73d-00155d000105 https://dk.um.si/Dokument.php?lang=slv&id=186122 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0