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Title:Precipitacija in aktivnost transglutaminaze v superkritičnem ogljikovem dioksidu : diplomsko delo univerzitetnega študijskega programa I. stopnje
Authors:ID Globočnik, Maja (Author)
ID Leitgeb, Maja (Mentor) More about this mentor... New window
ID Primožič, Mateja (Comentor)
ID Kravanja, Gregor (Comentor)
Files:.pdf UN_Globocnik_Maja_2019.pdf (1,88 MB)
MD5: 8248309747812695F7F795E5D4D0100F
PID: 20.500.12556/dkum/1494961a-78df-4bf4-b80d-31a68c8d453c
 
Language:Slovenian
Work type:Bachelor thesis/paper
Typology:2.11 - Undergraduate Thesis
Organization:FKKT - Faculty of Chemistry and Chemical Engineering
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
Place of publishing:Maribor
Place of performance:Maribor
Publisher:[M. Globočnik]
Year of publishing:2019
Number of pages:X, 37 str.
PID:20.500.12556/DKUM-74278 New window
UDC:534.272.62(043.2)
COBISS.SI-ID:22608150 New window
NUK URN:URN:SI:UM:DK:X449ANKO
Publication date in DKUM:06.09.2019
Views:2620
Downloads:136
Metadata:XML DC-XML DC-RDF
Categories:KTFMB - FKKT
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Licences

License:CC BY-NC-ND 4.0, Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
Link:http://creativecommons.org/licenses/by-nc-nd/4.0/
Description:The most restrictive Creative Commons license. This only allows people to download and share the work for no commercial gain and for no other purposes.
Licensing start date:21.08.2019

Secondary language

Language:English
Title:Precipitation and activity of transglutaminase in supercritical carbon dioxide
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.
Keywords:transglutaminase, precipitation, cross-linked enzyme aggregates, supercritical carbon dioxide, magnetic suspension balance, solubility of carbon dioxide in enzyme


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