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Title:IMOBILIZACIJA β-GALAKTOZIDAZE NA MAGNETNE MAGHEMITNE NANODELCE IN ZAMREŽENJE LE-TEH V ENCIMSKE SKUPKE
Authors:ID Kramberger, Nastja (Author)
ID Leitgeb, Maja (Mentor) More about this mentor... New window
ID Vasić, Katja (Comentor)
Files:.pdf UN_Simenko_Nastja_2017.pdf (2,12 MB)
MD5: C080856198AA7E3793B229CAFAABE450
 
Language:Slovenian
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:FKKT - Faculty of Chemistry and Chemical Engineering
Abstract:Namen našega dela je bil pripraviti zamrežene encimske skupke (CLEA) in magnetne zamrežene encimske skupke (mCLEA) iz encima β-galaktozidaze. Doseči smo želeli čim višjo preostalo aktivnost encima in čim višjo učinkovitost imobilizacije. Zamreženje smo preizkušali v štirih obarjalnih reagentih (acetonu, etanolu, 1-propanolu in 2-propanolu), dodajali smo ogrodne proteine in mrežne povezovalce v različnih kombinacijah in koncentracijah. Eksperimentalno delo je obsegalo pripravo vzorcev (obarjanje in zamreženje), aktivnostni test in Bradfordovo metodo za določanje koncentracije proteinov. Proces smo optimirali pri konstantni koncentraciji encima (50 mg/ml) in konstantnem dodatku glutaraldehida (GA). Ugotovili smo, da je optimalna količina dodanega mrežnega povezovalca GA 1.5% (v/v) in da mrežni povezovalec pentaetilenheksamin (PEHA) negativno vpliva na zamreženje, razen če ga dodajamo v kombinaciji z ogrodnimi proteini. V tem primeru se preostala aktivnost zviša, medtem ko učinkovitost imobilizacije močno upade.
Keywords:biokataliza, β-galaktozidaza, imobilizacija encima, zamreženi encimski skupki, magnetni nanodelci
Place of publishing:Maribor
Publisher:[N. Šimenko]
Year of publishing:2017
PID:20.500.12556/DKUM-61506 New window
UDC:544.473:577.15(043.2)
COBISS.SI-ID:20442646 New window
NUK URN:URN:SI:UM:DK:UYSXI9DX
Publication date in DKUM:30.03.2017
Views:1718
Downloads:204
Metadata:XML DC-XML DC-RDF
Categories:KTFMB - FKKT
KTFMB - FKKT
:
KRAMBERGER, Nastja, 2017, IMOBILIZACIJA β-GALAKTOZIDAZE NA MAGNETNE MAGHEMITNE NANODELCE IN ZAMREŽENJE LE-TEH V ENCIMSKE SKUPKE [online]. Master’s thesis. Maribor : N. Šimenko. [Accessed 18 April 2025]. Retrieved from: https://dk.um.si/IzpisGradiva.php?lang=eng&id=61506
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Secondary language

Language:English
Title:IMMOBILIZATION OF BETA-GALACTOSIDASE ON MAGNETIC MAGHEMITE NANOPARTICLES AND THEIR CROSS LINKING INTO ENZYME AGGREGATES
Abstract:The purpose of our work was to create cross-linked enzyme aggregates (CLEA) and magnetic cross-linked enzyme aggregates (mCLEA) off enzyme β-galactosidase. The aim of our work was to achive the highest possible residual activity and maximize the efficiency of immobilization. The cross-linking was tested in four most optimal precipitation reagents (acetone, ethanol, 1-propanol, 2-propanol) while stability proteins and cross-linkers in various combinations and concentrations were added to the synthesis reaction. The experimental work included preparation of samples (precipitation and cross-linking), activity test and the Bradford protein assay for determination of proteins. The process was optimized at a constant concentration enzyme (50 mg/ml), and a constant addition of glutaraldehyde (GA). It was concluded that the optimal quantity of added cross-linker is 1.5% (v/v) and that the cross-linker pentaetilenheksamin (PEHA) negatively affects the cross-linking, unless it is added in combination with stability proteins. In this case, the residual activity is increased, while the efficacy of immobilization drops significantly.
Keywords:biocatalysis, β-galactosidase, enzyme immobilization, cross-linked enzyme aggregates, magnetic nanoparticles


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