Zamreženje lizocima v encimske skupke

diplomsko delo visokošolskega strokovnega študijskega programa I. stopnje

Maja Verdev (Author), Mateja Primožič (Mentor), Maja Leitgeb (Co-mentor), Gordana Hojnik Podrepšek (Co-mentor)

Abstract

V okviru diplomskega dela smo izvedli optimizacijo imobilizacije lizocima v obliki zamreženih encimskih skupkov (CLEAs). Postopek imobilizacije sestoji iz dveh korakov; obarjanja in zamreženja. V prvem delu eksperimentalnega dela smo proučevali vpliv različnih obarjalnih reagentov na aktivnost lizocima. Tako smo določili optimalen obarjalni reagent. V drugem delu smo uspešno oborjen encim v ustreznih obarjalnih reagentih imobilizirali v obliki CLEAs z mrežnim povezovalcem glutaraldehidom (GA). Lizocim smo imobilizirali v CLEAs z dodatkom govejega serumskega albumina (BSA) in albumina iz jajčnih beljakov (EA) v dveh različnih pufrih. Uporabili smo natrijev fosfatni pufer (SP) in kalijev fosfatni pufer (PP). Za nadaljnje eksperimente smo uporabili PP, saj je bila ob njegovi uporabi dosežena višja aktivnost encima lizocim in višja učinkovitost imobilizacije. V diplomskem delu smo proučili tudi vpliv različnih dodatkov GA in EA na aktivnost in učinkovitost imobilizacije lizocima. Cilj diplomskega dela je bil poiskati optimalne pogoje za sintezo CLEAs iz encima lizocim z najvišjo aktivnostjo imobiliziranega lizocima in učinkovitostjo imobilizacije. Stabilnost imobiliziranega encima v obliki CLEAs iz lizocima smo proučili tako, da smo encim večkrat uporabili za izvedbo encimsko katalizirane reakcije. Sintetizirane CLEAs iz lizocima smo okarakterizirali z uporabo vrstične elektronske mikroskopije, optičnega mikroskopa in Fourierove transformacijske infrardeče spektroskopije.

Keywords

Lizocim;encimska aktivnost;imobilizacija lizocima;stabilnost;diplomske naloge;

Data

Language:	Slovenian
Year of publishing:	2021
Typology:	2.11 - Undergraduate Thesis
Organization:	UM FKKT - Faculty of Chemistry and Chemical Engineering
Publisher:	[M. Verdev]
UDC:	604.4:577.15(043.2)
COBISS:	80437763
Views:	318
Downloads:	45
Average score:	0 (0 votes)
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Other data

Secondary language:	English
Secondary title:	Cross-linking of lysozyme into enzyme aggreagates
Secondary abstract:	Within the diploma work, we performed optimization of lysozyme immobilization in the form of cross-linked enzyme clusters (CLEAs). The immobilization process consists of two steps; precipitation and crosslinking. In the first part of the experimental work, we first studied the influence of different precipitating reagents on lysozyme activity. Thus, we determined the optimal precipitating reagent. In the second part, the successfully precipitated enzyme, in the appropriate precipitating reagents, was immobilized in the form of CLEAs with a network linker glutaraldehyde (GA). Lysozyme was immobilized in CLEAs by the addition of bovine serum albumin (BSA) and egg white albumin (EA) in two different buffers. Sodium phosphate buffer (SP) and potassium phosphate buffer (PP) were used. PP was used for further experiments, as higher lysozyme activity and higher immobilization efficiency were achieved with its use. In the diploma work we also studied the influence of different additives GA and EA on the activity and efficiency of lysozyme immobilization. The aim of the thesis was to find the optimal conditions for the synthesis of CLEAs from the enzyme lysozyme with the highest activity of immobilized lysozyme and the efficiency of immobilization. The stability of the immobilized enzyme in the form of CLEAs from lysozyme was studied by repeated use of the immobilized enzyme to perform an enzyme-catalyzed reaction. Synthesized CLEAs from lysozyme were characterized using scanning electron microscopy, optical microscopy, and Fourier transform infrared spectroscopy.
Secondary keywords:	Lysozyme;CLEAs;enzyme activity;lysozyme immobilization;stability;
Type (COBISS):	Bachelor thesis/paper
Thesis comment:	Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Pages:	X, 40 str.
ID:	13208468