PRIPRAVA MAGNETNIH ZAMREŽENIH ENCIMSKIH SKUPKOV IZ β- GALAKTOZIDAZE

Kupnik, Kaja

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Title:	PRIPRAVA MAGNETNIH ZAMREŽENIH ENCIMSKIH SKUPKOV IZ β- GALAKTOZIDAZE
Authors:	ID Kupnik, Kaja (Author) ID Leitgeb, Maja (Mentor) More about this mentor... ID Vasić, Katja (Comentor)
Files:	UN_Jeromel_Kaja_2016.pdf (1,99 MB) MD5: 81DDF57296E85F258ED1495FBEF3507B
Language:	Slovenian
Work type:	Bachelor thesis/paper
Typology:	2.11 - Undergraduate Thesis
Organization:	FKKT - Faculty of Chemistry and Chemical Engineering
Abstract:	Namen diplomskega dela je bil pripraviti aktivne zamrežene encimske skupke (CLEAs) in magnetne zamrežene encimske skupke (mCLEAs) iz encima β-galaktozidaze ter pri tem doseči čim višjo učinkovitost imobilizacije in preostanek aktivnosti encima β-galaktozidaze. Pri tem smo optimirali parametre, da smo prišli do čim boljših rezultatov. Priprava CLEAs in mCLEAs je bila sestavljena iz postopka obarjanja in zamreženja. Pri obarjanju smo med drugim preučevali, katera organska topila so se izkazala kot najboljši obarjalni reagenti. Zamreženje pa smo optimirali s spreminjanjem časa in temperature. Ugotovili smo, da so najboljši obarjalni reagenti za pripravo CLEAs etanol, aceton, 1-propanol in 2-propanol, za pripravo mCLEAs pa etanol, 1-propanol in 2-propanol. Optimalni pogoji za pripravo CLEAs in mCLEAs so koncentracija encima β-galaktozidaze 50 mg/ml in 2-urno zamreženje pri 10°C ob dodatku 1,5% (v/v) mrežnega povezovalca glutaraldehida (GA). Ugotovili smo tudi, da so CLEAs in mCLEAs pri 2-urni izpostavljenosti na 50°C še stabilni, medtem ko pri izpostavljenosti na 70°C njihova aktivnost upade.
Keywords:	β-galaktozidaza, zamreženi encimski skupki, mrežni povezovalec, obarjanje, zamreženje, imobilizacija
Place of publishing:	Maribor
Publisher:	[K. Jeromel]
Year of publishing:	2016
PID:	20.500.12556/DKUM-62410
UDC:	543.645.4(043.2)
COBISS.SI-ID:	20367638
NUK URN:	URN:SI:UM:DK:QVC9BNGD
Publication date in DKUM:	04.10.2016
Views:	2070
Downloads:	295
Metadata:
Categories:	KTFMB - FKKT
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Secondary language

Language:	English
Title:	PREPARATION OF MAGNETIC CROSS-LINKED ENZYME AGGREGATES FROM β- GALACTOSIDASE
Abstract:	The purpose of the diploma work was to prepare cross-linked enzyme aggregates (CLEAs) as well as magnetic cross-linked enzyme aggregates (mCLEAs) from the enzyme β-galactosidase and achieve the highest efficiency of immobilization and the remaining activity of the enzyme β-galactosidase. With this process we optimized the parameters to achieve the best possible results. The preparation on CLEAs and the mCLEAs consisted of two procedures: precipitation and cross-linking. With the first we were examining which organic solvents worked out most beneficially. We optimized the cross-linking with variations in time and temperature. We determined that the best precipitated reagents for the preparation of CLEAs are ethanol, acetone, 1-propanol and 2-propanol and for the preparation of mCLEAs are ethanol 1-propanol and 2-propanol. The most optimal conditions for the preparation of CLEAs and mCLEAs occur with the concentration of the enzyme β-galactosidase 50 mg/ml, at a 2 hours cross-linking at 10°C with adding 1,5% (v/v) cross-linking agent glutaraldehyde (GA). We also realized that CLEAs and mCLEAs still remain stable when exposed to 50°C for 2 hours, but they lose their stability at 70°C.
Keywords:	β-galactosidase, cross-linked enzyme aggregates, cross-linking agent, precipitation, cross-linking, immobilisation

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