Koimobilizacija alkohol dehidrogenaze in kofaktorja ß-NAD na modificirane magnetne nanodelce

magistrsko delo

Eva Voda (Author), Maja Leitgeb (Mentor), Katja Heržič (Co-mentor)

Abstract

Encim ADH je precej nestabilen, kar omeji njegovo uporabo v industriji, zato je primarni cilj izboljšanje stabilnosti le-tega. To lahko dosežemo z imobilizacijo encima na ustrezne nosilce. S tem ne samo izboljšamo stabilnost, ampak tudi omogočimo odstranitev encima iz reakcijskega medija in zagotovimo ponovno uporabo, s čemer zelo zmanjšamo stroške reakcije. Magistrsko delo prikazuje študijo koimobilizacije encima alkohol dehidrogenaze (ADH) in kofaktorja β-nikotinamid adenin dinukleotida (β-NAD) na magnetne nanodelce, modificirane s karboksimetil dekstranom (CMD). Magnetne nanodelce smo pred koimobilizacijo aktivirali z uporabo mrežnega povezovalca epiklorohidrina (EClH). Pred nadaljnjimi študijami smo morali najti optimalne pogoje imobilizacije, in sicer s spremljanjem aktivnosti ADH ter učinkovitosti imobilizacije. Učinkovitost imobilizacije smo določili s pomočjo Bradfordove metode za določevanje koncentracije totalnih proteinov. Rezultati magistrskega dela kažejo, da z imobilizacijo ADH zelo izboljšamo njeno termično stabilnost, je primerna za shranjevanje pri 4 °C in dobro ohrani aktivnost pri ponovni uporabi.

Keywords

koimobilizacija;alkohol dehidrogenaza;β-NAD kofaktor;magnetni nanodelci;karboksimetil dekstran;magistrske naloge;

Data

Language:	Slovenian
Year of publishing:	2022
Typology:	2.09 - Master's Thesis
Organization:	UM FKKT - Faculty of Chemistry and Chemical Engineering
Publisher:	[E. Voda]
UDC:	544.567:620.3(043.2)
COBISS:	116182019
Views:	71
Downloads:	11
Average score:	0 (0 votes)
Metadata:

Other data

Secondary language:	English
Secondary title:	Coimobilization of alcohol dehydrogenase and ß-NAD cofactor to modified magnetic nanoparticles
Secondary abstract:	The enzyme alcohol dehydrogenase is rather unstable which limits its use in industry; therefore there is the desire to improve its stability. This can be achieved by immobilizing the enzymes onto appropriate carriers. This does not only improve the stability of the enzyme; it also allows enzymes to be removed from the reaction stream and then be reused, thus greatly reducing the reaction costs. The master's thesis presents a study of the co-mobilization of enzyme alcohol dehydrogenase (ADH) and β-nicotineamide adenine dinucleotide (β-NAD) cofactor onto magnetic nanoparticles modified with carboxymethyl dextran (CMD). Magnetic nanoparticles need to be activated before the co-mobilization which is achieved using a cross-linking agent epichlorohydrin (EClH). Prior to further studies, we had to find optimal conditions for immobilization by monitoring the activity of alcohol dehydrogenase and the immobilization efficiency. The immobilization efficiency was determined using the Bradford method. The results of the master's thesis show that co-mobilization of ADH greatly improves its thermal and storage stability and has a good residual activity after reusing it.
Secondary keywords:	coimmobilization;alcohol dehydrogenase;β-NAD cofactor;magnetic nanoparticles;CM-dextran;
Type (COBISS):	Master's thesis/paper
Thesis comment:	Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Pages:	1 spletni vir (1 datoteka PDF (X, 46 f.))
ID:	15695107