Stabilnost lizocima pri ekstremnih pogojih

diplomsko delo visokošolskega strokovnega študijskega programa I. stopnje

Lucija Dolinšek (Author), Maja Leitgeb (Mentor), Mateja Primožič (Co-mentor)

Abstract

V diplomskem delu smo preučevali vpliv SC CO2 na aktivnost prostega in imobiliziranega encima lizocim ob spreminjanju različnih parametrov, kot so tlak, temperatura in čas izpostavitve. Za primerjavo smo proučili tudi vpliv temperature in časa izpostavitve na stabilnost obeh oblik encima pri atmosferskem tlaku. Analizirali smo prosti in imobilizirani encim brez nosilca v obliki CLEAs. Namen diplomske naloge je bilo določiti optimalne pogoje za izpostavitev prostega encima in CLEAs v SC CO2 z morebitno hiperaktivacijo encima. Koncentracijo celokupnih proteinov smo določili z Bradfordovo metodo. S pomočjo UV-Vis spektrofotometra smo prostemu in imobiliziranemu encimu določili prisotnost celokupnih proteinov in aktivnost lizocima. Delež vode v vzorcih smo proučevali s pomočjo tehtanja mikrocentifugirk z vzorcem, in sicer pred in po izpostavitvi v SC CO2. Rezultati prikazujejo ugoden vpliv SC CO2 na aktivnost prostega in imobiliziranega encima, saj se je aktivnost obeh encimov po izpostavitvi v SC CO2 zvišala. Ugotovili smo, da je imobiliziran encim bolj stabilen kot prosti encim, kar se še posebej vidi pri ekstremnih pogojih (višja temperatura in tlak izpostavitve) in daljšem času izpostavitve v SC CO2. Z višanjem temperature in tlaka izpostavitve je prišlo do upada deleža vode v CLEAs.

Keywords

lizocim;superkritični ogljikov dioksid;zamreženi encimski skupki;aktivnost in stabilnost lizocima;diplomske naloge;

Data

Language:	Slovenian
Year of publishing:	2021
Typology:	2.11 - Undergraduate Thesis
Organization:	UM FKKT - Faculty of Chemistry and Chemical Engineering
Publisher:	[L. Dolinšek]
UDC:	544.032.1:577.15(043.2)
COBISS:	72407043
Views:	606
Downloads:	96
Average score:	0 (0 votes)
Metadata:

Other data

Secondary language:	English
Secondary title:	Stability of lysozyme under extreme conditions
Secondary abstract:	In this thesis, we studied the influence of SC CO2 on the activity of free and immobilized enzyme Lysozyme by changing various parameters such as pressure, temperature, and exposure time. For comparison, we also studied the effect of temperature and exposure time on the stability of both forms of the enzyme at atmospheric pressure. Free and immobilized carrier-free enzyme in the form of CLEAs was analysed. The purpose of the thesis was to determine the optimal conditions for exposure of free enzyme and CLEAs in SC CO2 with possible hyperactivation of the enzyme. The concentration of total proteins was determined by the Bradford method. Using an UV-Vis spectrophotometer, the presence of total proteins and lysozyme activity was determined fort the free and immobilized enzyme. The water content in the samples was studied by weighing the sample microcentrifuges tubes before and after exposure to SC CO2. The results show a beneficial effect of SC CO2 on the activity of the free and immobilized enzyme, as the activity of both enzymes increased after exposure to SC CO2. We found that the immobilized enzyme is more stable than the free enzyme, which is especially seen under extreme conditions (higher temperature and pressure exposure) and longer exposure time in SC CO2. As the temperature and pressure exposure increased, the proportion of water in the CLEAs decreased.
Secondary keywords:	Lysozyme;supercritical carbon dioxide;cross-linked enzyme aggregates;lysozyme activity and stability;
Type (COBISS):	Bachelor thesis/paper
Thesis comment:	Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Pages:	IX, 44 str.
ID:	13059935