20.500.12556/DKUM-60906 PRIPRAVA MAGNETNIH ZAMREŽENIH ENCIMSKIH SKUPKOV IZ ENCIMA CELULAZA PREPARATION OF MAGNETIC CROO-LINKED ENZYME AGGREGATES OF THE CELLULASE Namen magistrskega dela je določiti vpliv posameznih sinteznih parametrov na fizikalne in kemijske lastnosti zamreženih encimskih skupkov in magnetnih zamreženih encimskih skupkov iz encima celulaze in narediti primerjavo med CLEAs in mCLEAs. Iz encima celulaze smo vzporedno pripravili zamrežene encimske skupke in magnetne zamrežene encimske skupke in izvedli optimizacijo parametrov, ki vplivajo na aktivnost, učinkovitost in stabilnost imobilizacije. Preučevali smo vpliv obarjalnih reagentov, mrežnih povezovalcev, vpliv koncentracije glutaraladehida v kombinaciji z različnimi obarjalnimi reagenti na aktivnost in učinkovitost CLEAs in mCLEAs, temperaturo zamreženja, vpliv koncentracije encima, vpliv dodatkov in izpiranja z puferno raztopino ter vpliv koncentracije natrijevega cianoborohidrida na aktivnost in stabilnost imobilizacije. Uspešno smo sintetizirali celulazno CLEAs in mCLEAs, dobljene relativne aktivnosti za CLEAs so 86% in za mCLEAs 91%. Končna aktivnost po spiranju z vodo je večja za mCLEAs kot za CLEAs, ker imajo amino-funkcionalizirani magnetni nano delci dodatno povezovalno vlogo v postopku zamreženja. Izvedene so meritve stabilnosti na različnih temperaturah in možnost ponovne uporabe encima. The aim of this graduate thesis is to determine the influence of the synthesis parameters, such as different precipitants, additives, cross-linker and other parameters on the physical and chemical characteristics of cross-linked enzyme aggregates and magnetic cross-linked enzyme aggregates of the enzyme cellulase and comparisson of the characteristics of CLEAs and mCLEAs. The cross-linked enzyme aggregates CLEAs and magnetic cross-linked enzyme aggregates mCLEAs were prepared of the enzyme cellulase and optimization of the various parameters were performed paralelly. The effect of different precipitants, cross-linking agents, the impact of concentration glutaraldehyde in combination with a different precipitants to the activity and immobilisation efficiency of CLEAs and mCLEAs, the temperature of cross-linking, the effect of the concentration of the enzyme, the effect of additives, buffer washing, the influence of the concentration of sodium cyanoborohydride to the activity and stability were investigated. The cellulase CLEAs and mCLEAs were prepared successfully, relative activities for CLEAs 86% and 91% for mCLEAs were obtained. The final activity is higher for mCLEAs because of presence the amino-functionalized magnetic nanoparticles. Measurements of stability at different temperatures and reusability of the CLEAs and mCLEAs were performed. celulaza zamreženi encimski skupki magnetni zamreženi encimski skupki magnetni nanodelci maghemita mrežni povezovalec cellulase cross-linked enzyme aggregates magnetic cross-linked enzyme aggregates maghemite magnetic nanoparticles cross-linking agent true true false [D. Ifko] Slovenski jezik Angleški jezik Magistrsko delo 2016-07-08 09:47:41 2016-10-13 12:21:39 2022-06-25 03:06:38 2019-07-11 12:02:27 2016 0 Maribor 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 577.151(043) 19876374 URN:SI:UM:DK:YZBWKR2O MAG_Ifko_Dusica_2016.pdf MAG_Ifko_Dusica_2016.pdf 1 AE0988FA0042FA3C26828AC2FC3D3A33 6a1675b268f54b3764d7229d3074440f12cdf73f2fe9bc013c1a87bd3a5bd316 c485de86-7c0d-11eb-bb7a-00155d0001ca https://dk.um.si/Dokument.php?lang=slv&id=107433 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0