20.500.12556/DKUM-68276 Imobilizacija lakaze v zamrežene encimske skupke (CLEAs) Immobilization of laccase in the cross-linked enzyme aggregates (CLEAs) Diplomsko delo opisuje sintezo zamreženih encimskih skupkov iz encima lakaze (CLEAs). Sinteza CLEAs delcev iz lakaze je potekala po določenem postopku, pri katerem sta najpomembnejši fazi obarjanje in zamreženje. Raztopino encima lakaze smo obarjali v različnih obarjalnih reagentih, najuspešnejša sta bila obarjalna reagenta etanol, 1-propanol in 2-propanol. Oborjeni encim smo zamreževali z mrežnim povezovalcem glutaraldehidom. Zamreženim encimskim skupkom smo določevali učinkovitost imobilizacije in preostalo aktivnost imobiliziranega encima v primerjavi s prostim encimom. Pri postopku imobilizacije smo z namenom, da bi dosegli čim višjo učinkovitost imobilizacije ter čim višjo preostalo aktivnost imobiliziranega encima, spreminjali parametre, kot so volumski deleži glutaraldehida, čas zamreženja, vpliv ogrodnih proteinov govejega seruma albumina (BSA) in jajčnega albumina (EA), volumski deleži natrijevega cianoborohidrida in temperaturo zamreženja. Vse vzorce smo tudi dvakrat spirali, z namenom da bi odstranili nezamrežen encim. The diploma work describes synthesis of cross-linked enzyme aggregates (CLEAs) of laccase enzyme. Synthesis of CLEAs laccase was carried out according to a particular process in which percipitation and cross-linking are the most important stages. The solution of the laccase enzyme was precipitated in various precipitation reagents, the most successful were ethanol, 1-propanol and 2-propanol. The precipitated enzyme was cross-linked with the cross-linker glutaraldehyde. The immobilization efficiency and the residual specific activity of immobilized enzyme as cross-linked enzyme aggregates was determined in comparison with free enzyme. Parameters, such as volume fraction of glutaraldehyde, cross-linking time, influence of stability proteins bovine serum albumin (BSA) and egg albumin (EA), volume fraction of sodium cyanoborohydride and the temperature of cross-linking, have been optimized to achieve the highest immobilization efficiency and the highest residual specific activity of the cross-linked enzyme. All the samples were also washed two times to remove the noncross-linked enzyme. Lakaza imobilizacija zamreženi encimski skupki obarjanje zamreženje glutaraldehid encimska aktivnost Laccase immobilization cross-linked enzyme aggregates precipitation cross-linking glutaraldehyde enzyme activity true true false [M. Petek] Slovenski jezik Angleški jezik Diplomsko delo/naloga 2017-09-14 13:07:22 2017-11-22 08:30:27 2022-08-01 15:05:20 2019-07-11 16:47:41 2017 0 Maribor 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 544.478.3(043.2) 21062166 URN:SI:UM:DK:JVMQ3FEQ UN_Petek_Mihaela_2017.pdf UN_Petek_Mihaela_2017.pdf 1 63DF079D0DE232015B048FE5E389102D e1d70d66d7d4b6b0ec5e5c2c755d274d2e2808a351a92e6a2563794e94c4546d 534e0cd3-7c0f-11eb-bb7a-00155d0001ca 20.500.12556/dkum/9764e1a9-ef88-4904-87ab-803588f7ffb2 https://dk.um.si/Dokument.php?lang=slv&id=118640 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0