20.500.12556/DKUM-74278 Precipitacija in aktivnost transglutaminaze v superkritičnem ogljikovem dioksidu diplomsko delo univerzitetnega študijskega programa I. stopnje Precipitation and activity of transglutaminase in supercritical carbon dioxide 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. 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. transglutaminaza precipitacija zamreženi encimski skupki superkritični ogljikov dioksid magnetna suspenzijska tehtnica topnost ogljikovega dioksida v encimu transglutaminase precipitation cross-linked enzyme aggregates supercritical carbon dioxide magnetic suspension balance solubility of carbon dioxide in enzyme true true false [M. Globočnik] Slovenski jezik Angleški jezik Diplomsko delo/naloga 2019-08-21 12:00:39 2019-09-06 08:40:42 2022-08-09 08:33:55 2019-09-26 03:11:39 2019 0 Maribor 0 Maribor X, 37 str. 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 534.272.62(043.2) 22608150 URN:SI:UM:DK:X449ANKO UN_Globocnik_Maja_2019.pdf UN_Globocnik_Maja_2019.pdf 1 8248309747812695F7F795E5D4D0100F 05c059a78742265c7bc53ae76d8ae5f7eb866df3328b64d22fbe38c3a39b5f5b e4b69f6b-7c10-11eb-bb7a-00155d0001ca 20.500.12556/dkum/1494961a-78df-4bf4-b80d-31a68c8d453c https://dk.um.si/Dokument.php?lang=slv&id=137065 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0