Precipitacija in aktivnost transglutaminaze v superkritičnem ogljikovem dioksidu

diplomsko delo univerzitetnega študijskega programa I. stopnje

Maja Globočnik (Author), Maja Leitgeb (Mentor), Mateja Primožič (Co-mentor), Gregor Kravanja (Co-mentor)

Abstract

Diplomsko delo obravnava obarjanje encima transglutaminaze (TG), zamreženje TG in optimiranje sinteze zamreženih encimskih skupkov TG (CLEAs TG) v superkritičnem ogljikovem dioksidu (SC CO2) ter določanje aktivnosti TG. Najprej smo preliminarno z magnetno suspenzijsko tehtnico (MSB) izmerili topnost in določili difuzivnost CO2 v encimu. Nato smo encim obarjali in zamreževali v SC CO2 pri različnih pogojih. Preverjali smo vpliv pH in tlaka na aktivnost oborjenega, nezamreženega encima. Pri postopku sinteze CLEAs TG v SC CO2 smo proučili vpliv tlaka in količine dodanega mrežnega povezovalca glutaraldehida (GA) na učinkovitost imobilizacije in preostalo aktivnost CLEAs TG. Nazadnje smo proučili še vpliv dodatka ogrodnega proteina albumina iz govejega seruma (BSA) na učinkovitost imobilizacije in preostalo aktivnost sintetizirane CLEAs TG. Rezultati, dobljeni s pomočjo MSB, so pokazali dobro topnost CO2 v encimu TG. Najvišji difuzijski koeficient za sistem CO2/TG smo določili pri tlaku 10 MPa in temperaturi 37 °C. Pri tem tlaku sta najvišjo preostalo aktivnost ohranila tako nezamreženi encim (obarjanje) kot zamreženi encim CLEAs TG (z dodatkom 5 % (v/v) GA). Ob dodatku ogrodnega proteina BSA smo dobili večje in kompaktnejše CLEAs TG.

Keywords

transglutaminaza;precipitacija;zamreženi encimski skupki;superkritični ogljikov dioksid;magnetna suspenzijska tehtnica;topnost ogljikovega dioksida v encimu;diplomske naloge;

Data

Language:	Slovenian
Year of publishing:	2019
Typology:	2.11 - Undergraduate Thesis
Organization:	UM FKKT - Faculty of Chemistry and Chemical Engineering
Publisher:	[M. Globočnik]
UDC:	534.272.62(043.2)
COBISS:	22608150
Views:	1443
Downloads:	95
Average score:	0 (0 votes)
Metadata:

Other data

Secondary language:	English
Secondary title:	Precipitation and activity of transglutaminase in supercritical carbon dioxide
Secondary abstract:	The matter of this diploma thesis is precipitation of enzyme transglutaminase (TG), cross-linking of tranglutaminase (TG), optimising the synthesis of cross-linked enzyme aggregates of TG (CLEAs TG) in supercritical CO2, and defining the activity of TG. As a preliminary research, we have used magnetic suspension balance (MSB) to define solubility and diffusivity of CO2 in enzyme. Then we precipitated and cross-linked the enzyme in SC CO2 at different conditions. We have tested the effect of pH and pressure on the activity of precipitated, non-cross-linked enzyme. At the procedure of CLEAs TG, we have checked the effect of pressure and the amount of cross-linking agent glutaraldehyde (GA) on the immobilisation and residual activity. Lastly, we have investigated the effect of carrier protein albumin derived from cows (BSA) on the immobilisation and residual activity of synthesized CLEAs TG. The MSB results have shown good solubility of CO2 in enzyme TG. Diffusion coefficient for the system CO2/TG has reached its maximum at pressure 10 MPa and temperature 37 °C. At the same pressure, both non-cross-linked (precipitation) and cross-linked enzyme CLEAs TG (with added 5 % (v/v) of cross-linker) preserved the highest residual activity. BSA has helped in achieving bigger and more compact CLEAs TG.
Secondary keywords:	transglutaminase;precipitation;cross-linked enzyme aggregates;supercritical carbon dioxide;magnetic suspension balance;solubility of carbon dioxide in enzyme;
Type (COBISS):	Bachelor thesis/paper
Thesis comment:	Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Pages:	X, 37 str.
ID:	11204842