Imobilizacija lakaze v magnetne zamrežene encimske skupke (mCLEAs)

diplomsko delo univerzitetnega študijskega programa I. stopnje

Nina Šenekar (Avtor), Maja Leitgeb (Mentor), Katja Heržič (Komentor)

Povzetek

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.

Ključne besede

lakaza;glutaraldehid;imobilizacija;magnetni nanodelci;magnetni zamreženi encimski skupki;diplomske naloge;

Podatki

Jezik:	Slovenski jezik
Leto izida:	2017
Tipologija:	2.11 - Diplomsko delo
Organizacija:	UM FKKT - Fakulteta za kemijo in kemijsko tehnologijo
Založnik:	[N. Šenekar]
UDK:	620.3:577.15(043.2)
COBISS:	21051414
Št. ogledov:	846
Št. prenosov:	103
Ocena:	0 (0 glasov)
Metapodatki:

Ostali podatki

Sekundarni jezik:	Angleški jezik
Sekundarni naslov:	Immobilization of lacasse in the magnetic cross-linked enzyme aggregates (mCLEAs)
Sekundarni povzetek:	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.
Sekundarne ključne besede:	lacasse;glutaldehyde;magnetic nanoparticles;immobilization;magnetic cross-linked enzyme aggregates;
URN:	URN:SI:UM:
Vrsta dela (COBISS):	Diplomsko delo/naloga
Komentar na gradivo:	Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Strani:	X, 32 str.
ID:	10860495