Vpliv superkritičnega ogljikovega dioksida na aktivnost imobilizirane transglutaminaze

magistrsko delo

Nika Kodba (Author), Mateja Primožič (Mentor), Maja Leitgeb (Co-mentor)

Abstract

V magistrskem delu smo raziskali vpliv superkritičnega ogljikovega dioksida (SC CO2) na aktivnost proste in imobilizirane transglutaminaze (TGM), ob spreminjanju različnih parametrov, ki lahko vplivajo na aktivnost encima kot so tlak, temperatura in čas izpostavitve v SC CO2. Pripravili smo dve obliki imobiliziranega encima in sicer z metodo obarjanja in zamreženja smo pripravili imobilizirano TGM brez nosilca (CLEAs) ter z metodo koprecipitacije imobilizirano TGM na magnetni nanonosilec, modificiran z aminosilanom (mCLEAs). Rezultati kažejo, da je TGM zamrežena v encimske skupke (CLEAs) stabilen katalizator ter primerna za izvedbo encimskih reakcij v SC CO2, saj smo pri optimalnih pogojih izpostavitve v SC CO2 (24 urna inkubacija, tlak 100 bar in temperatura 50 °C) dosegli 123 % preostale aktivnosti imobiliziranega encima. Kljub temu, da se je aktivnost CLEAs po izpostavitvi v SC CO2 za 24 h pri 100 bar povečala pa so bile aktivnosti CLEAs in mCLEAs po izpostavitvi pri atmosferskem tlaku pri enakih pogojih višje. Preostala aktivnost CLEAs pri atmosferskem tlaku je znašala 187 % in mCLEAs 139 %. Do podobnih ugotovitev smo prišli pri spreminjanju časa poteka encimske reakcije. Predpostavljamo, da SC CO2 povzroči konformacijske spremembe v proteinu, kadar je le ta imobiliziran in tako se zmanjša njegova aktivnost. Kljub manjši stabilnosti imobiliziranega encima v obliki CLEAs in mCLEas pa je bila možna njuna ponovna uporaba.

Keywords

transgultaminaza;superkritični ogljikov dioksid;zamreženi encimski skupki;magnetni nanodelci;aminosilan;magistrske naloge;

Data

Language:	Slovenian
Year of publishing:	2019
Typology:	2.09 - Master's Thesis
Organization:	UM FKKT - Faculty of Chemistry and Chemical Engineering
Publisher:	[N. Kodba]
UDC:	544.725(043.2)
COBISS:	22599446
Views:	745
Downloads:	101
Average score:	0 (0 votes)
Metadata:

Other data

Secondary language:	English
Secondary title:	The influence of supercritical carbon dioxide on the activity of immobilized transglutaminase
Secondary abstract:	In the contex of this master thesis we explored the influence of supercritical carbon dioxide (SC CO2) on free and immobilized transglutaminase activity (TGM), while changing various parameters that influence the activity of the enzyme such as pressure, temperature and reaction time. Two forms of immobilized enzyme were prepared. Using the precipitation and crosslinking method a immobilized TGM without carrier (CLEA) was prepared, additionally with the co-precipitation method a immobilized TGM was applied to a magnetic nanocarrier modified by aminosilane (mCLEAs). The results show that TGM crosslinked in enzymatic aggregates (CLEAs) is a stable catalyst and suitable for the implementation of enzymatic reactions in SC CO2, under optimum conditions of exposure to SC CO2 (24 hour incubation, 100 bar pressure and 50 °C temperature), we achieved 123% of the remaining activity. Despite the fact that CLEAs activity after exposure in SC CO2 for 24 h at 100 bar increased, CLEAs and mCLEAs activities at atmospheric pressure were higher. The remaining CLEAs activity at atmospheric pressure was 187% and 139% for mCLEAs. Similar findings were obtained in changing the duration of the enzyme reaction. We assumed that SC CO2 causes conformational changes in protein when it is immobilized and thus reduces its activity. Despite the lower stability of the immobilized enzyme, their reuse was possible.
Secondary keywords:	transglutaminase;supercritical carbon dioxide;cross-linked enzyme aggregates;magnetic nanoparticles;aminosilane;
URN:	URN:SI:UM:
Type (COBISS):	Master's thesis/paper
Thesis comment:	Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Pages:	X, 62 str.
ID:	11163470