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Title:
Imobilizacija lakaze v magnetne zamrežene encimske skupke (mCLEAs)
Authors:
ID
Šenekar, Nina
(Author)
ID
Leitgeb, Maja
(Mentor)
More about this mentor...
ID
Vasić, Katja
(Comentor)
Files:
UN_Senekar_Nina_2017.pdf
(1,45 MB)
MD5: CEC5625E3D76C878C3964F51222FE75A
PID:
20.500.12556/dkum/34d43aa0-f55f-448e-a87b-5be982b8d73f
Language:
Slovenian
Work type:
Bachelor thesis/paper
Typology:
2.11 - Undergraduate Thesis
Organization:
FKKT - Faculty of Chemistry and Chemical Engineering
Abstract:
Pripravili smo zamrežene magnetne encimske skupke (mCLEAs) iz lakaze. Metoda sestoji iz dveh delov: obarjanje encima z ustreznim obarjalnim reagentom in imobilizacija dobljenega agregata v zamrežene encimske skupke z zamreževalcem glutaraldehidom. S centrifugiranjem smo produkt ločili na pelet in supernatant. Supernatant smo uporabili za test koncentracije proteinov, na peletu pa smo izmerili preostalo aktivnost lakaze. Namen diplomske naloge je bil pripraviti mCLEAs z najvišjo preostalo aktivnostjo lakaze in učinkovitostjo imobilizacije. Preizkušali smo vpliv naslednjih parametrov: koncentracija magnetnih nanodelcev, koncentracijo glutaraldehida, količina NaBH3CN, temperatura, čas ter koncentracija encima. Raztopini encima smo dodajali ogrodne proteine: albumin iz govejega seruma (BSA) ter albumin iz kokošjih jajc (EA). mCLEAs z visoko preostalo aktivnostjo smo pripravili z uporabo 10 mg magnetnih nanodelcev v 100 µL raztopine lakaze koncentracije 20 mg/ml. Obarjali smo z 900 µL obarjalnega reagenta 2-propanol. Zamreževali smo z 15% (v/v) GA (150 µL) in dodali 50 µL NaBH3CN. Zamreževanje se je vršilo tri ure s hlajenjem.
Keywords:
lakaza
,
glutaraldehid
,
imobilizacija
,
magnetni nanodelci
,
magnetni zamreženi encimski skupki
Place of publishing:
Maribor
Publisher:
[N. Šenekar]
Year of publishing:
2017
PID:
20.500.12556/DKUM-67715
UDC:
620.3:577.15(043.2)
COBISS.SI-ID:
21051414
NUK URN:
URN:SI:UM:DK:WQE6N12W
Publication date in DKUM:
19.09.2017
Views:
1381
Downloads:
127
Metadata:
Categories:
KTFMB - FKKT
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:
ŠENEKAR, Nina, 2017,
Imobilizacija lakaze v magnetne zamrežene encimske skupke (mCLEAs)
[online]. Bachelor’s thesis. Maribor : N. Šenekar. [Accessed 23 January 2025]. Retrieved from: https://dk.um.si/IzpisGradiva.php?lang=eng&id=67715
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Licences
License:
CC BY-ND 4.0, Creative Commons Attribution-NoDerivatives 4.0 International
Link:
http://creativecommons.org/licenses/by-nd/4.0/
Description:
Under the NoDerivatives Creative Commons license one can take a work released under this license and re-distribute it, but it cannot be shared with others in adapted form, and credit must be provided to the author.
Licensing start date:
28.08.2017
Secondary language
Language:
English
Title:
Immobilization of lacasse in the magnetic cross-linked enzyme aggregates (mCLEAs)
Abstract:
Magnetic cross-linked enzyme aggregates (mCLEAs) of lacasse were prepared. The method consists of two parts: precipitation of enzyme with an appropriate precipitant agent and immobilization of the resulting aggregate into a cross-linked enzyme aggregates with the glutaraldehyde as cross-linker. After centrifugation, product was separated on pellet and supernatant. Supernatant was used to test protein concentration and pellet was used to measuse the residual activity of cross-linked lacasse. The purpose of diploma work was to prepare mCLEAs with the highest residual activity and immobilization efficiency. We tested the influence of the following parameters: concentration of magnetic nanoparticles, amount ob NaBH3CN, concentration of glutaraldehyde, time, temperature and concentration of lacasse. Proteins bovine serum albumin (BSA) and chicken egg albumin (EA) were added to enzyme solution. mCLEAs with highest residual activity were prepared using 10 mg of magnetic nanoparticles in 100 µL enzyme solution concentration of 20 mg/ml. 900 µL of 2-propanol was used as the most suitable precipitation reagent. Cross-linking was performed with 15% (v/v) GA. We also added 50 µL NaBH3CN. Cross-linking lasted for three hours with cooling.
Keywords:
lacasse
,
glutaraldehyde
,
magnetic nanoparticles
,
immobilization
,
magnetic cross-linked enzyme aggregates
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