Imobilizacija ß-galaktozidaze na magnetne maghemitne nanodelce in zamreženje le-teh v encimske skupke

diplomsko delo univerzitetnega študijskega programa I. stopnje

Nastja Šimenko (Avtor), Maja Leitgeb (Mentor), Katja Heržič (Komentor)

Povzetek

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.

Ključne besede

biokataliza;beta-galaktozidaza;imobilizacija encimov;zamreženi encimski skupki;magnetni nanodelci;diplomske naloge;

Podatki

Jezik:	Slovenski jezik
Leto izida:	2016
Tipologija:	2.11 - Diplomsko delo
Organizacija:	UM FKKT - Fakulteta za kemijo in kemijsko tehnologijo
Založnik:	[N. Šimenko]
UDK:	544.473:577.15(043.2)
COBISS:	20442646
Št. ogledov:	1224
Št. prenosov:	128
Ocena:	0 (0 glasov)
Metapodatki:

Ostali podatki

Sekundarni jezik:	Angleški jezik
Sekundarni naslov:	IMMOBILIZATION OF BETA-GALACTOSIDASE ON MAGNETIC MAGHEMITE NANOPARTICLES AND THEIR CROSS LINKING INTO ENZYME AGGREGATES
Sekundarni povzetek:	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.
Sekundarne ključne besede:	biocatalysis;beta-galactosidase;enzyme immobilization;cross-linked enzyme aggregates;magnetic nanoparticles;
URN:	URN:SI:UM:
Vrsta dela (COBISS):	Diplomsko delo/naloga
Komentar na gradivo:	Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Strani:	XII, 40 str.
ID:	9587773