20.500.12556/DKUM-68212 Priprava magnetnih zamreženih encimskih skupkov (cleas) iz transglutaminaze Preparation of magnetic cross-linked enzyme aggregates from transglutaminase Namen diplomskega dela je bil doseči čim višjo učinkovitost imobilizacije in čim višjo preostalo aktivnost imobiliziranega encima transglutaminaze (TG) z optimiranjem parametrov pri pripravi zamreženih encimskih skupkov (CLEAs) in pripravi magnetnih zamreženih encimskih skupkov (mCLEAs). Encim TG je služil za pripravo CLEAs in mCLEAs, ki smo ju izvedli vzporedno, ter tako pripravljenim vzorcem spreminjali parametre, ki so vplivali na stabilnost, učinkovitost in aktivnost imobiliziranih skupkov. Proučevali smo vpliv obarjalnih reagentov, vpliv koncentracije glutaraladehida (GA), mrežnih povezovalcev, temperaturo zamreževanja, vpliv podaljšanega časa zamreževanja, vpliv koncentracije encima in vpliv koncentracije natrijevega cianoborohidrida na aktivnost in stabilnost imobiliziranega encima. Rezultati prikazujejo, da je bila aktivnost CLEAs najvišja, ko smo obarjali encim TG s koncentracijo 100 mg/ml v obarjalnem reagentu 2-propanolu ob dodatku 10 % (v/v) mrežnega povezovalca GA in 200 µl natrijevega cianoborohidrida. Ugotovili smo tudi, da je bila dosežena najboljša aktivnost, z enakim obarjalnim reagentom in enako količino mrežnega povezovalca GA, kot smo ju uporabili pri CLEAs, mCLEAs z koncentracijo encima TG 200 mg/ml in 150 µl natrijevega cianoborohidrida. The purpose od this work was to achive the highest efficiency of immobilization and the highest remaining activity of the enzyme transglutaminase (TG). With optimization of the parameters we prepared cross-linked enzyme aggregates (CLEAs) and magnetic cross-linked enzyme aggregates (mCLEAs). Enzyme TG was used to prepare CLEAs and mCLEAs, which were carried out parallelly, and thus changing the paarameters that influenced the stabillity, efficiency and activity of the immobilized aggregates. We studied the impact of the precipitation reagents, the influence of the concentration of glutaraldehyde (GA), the cross-linking agent, the crosslinking temperature, the influence of the extended cross-linking time, the effect of the enzyme concentration and the influence of the concentration of sodium cyanoborohydride on the activity and stability of the immobilized enzyme. The results show that the CLEAs activity was highest when the TG enzyme was precipitated at a concentration of 100 mg/ml in the precipitating reagent 2-propanol, with the addition of 10 % (v/v) cross-linking agent GA and 200 µl of sodium cyanoborohydride. We also found that the highest activity was achived with the same precipitation agent and the same amount of cross-linking agent GA as used in CLEAs, mCLEAs with a concentration of TG 200 mg/ml and 150 µl of sodium cyanoborohydride. glutaraldehid obarjanje transglutaminaza zamreženi encimski skupki magnetni zamreženi encimski skupki glutaraldehyde precipitation transglutaminase cross-linked enzyme aggregates magnetic cross-linked enzyme aggregates true true false [N. Büdefeld] Slovenski jezik Angleški jezik Diplomsko delo/naloga 2017-09-13 03:34:26 2017-11-22 08:37:58 2022-08-01 15:01:07 2019-07-11 16:45:39 2017 0 Maribor 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 615.355:66.066.7(043.2) 21062678 URN:SI:UM:DK:JLN8DGC8 VS_Budefeld_Natalija_2017.pdf VS_Budefeld_Natalija_2017.pdf 1 77376A55CE02C6E104E38C720270F8DE ef66124c6093bfe4ffd9e2675e3818b3b693b0e7389f027b46121a560d9c4e45 62f6842a-7c0f-11eb-bb7a-00155d0001ca 20.500.12556/dkum/af0c2766-7f60-4e8b-a322-587cdab2e6ac https://dk.um.si/Dokument.php?lang=slv&id=119164 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0