Različne tehnike imobilizacije encima beta-galaktozidaze

magistrsko delo

Lucija Križnik (Author), Maja Leitgeb (Mentor), Mateja Primožič (Co-mentor)

Abstract

Namen magistrskega dela je primerjava kemijske in fizikalne metode imobilizacije encima ter imobilizacije encima brez nosilca, tj. v tako imenovane zamrežene encimske skupke. Za imobilizacijo smo uporabili encim ß-galaktozidazo, kateri razgrajuje disaharid laktozo na monosaharida, glukozo in galaktozo. V okviru magistrske naloge smo uspešno sintetizirali magnetne nanodelce maghemita, katere smo funkcionalizirali z aminoorganosilani in hitozanom. Encim smo kovalentno vezali na aminosilanske ter hitozanske magnetne nanodelce maghemita. Preizkusili smo tudi ujetje encima v netopnem gelu, za kar smo uporabili natrijev alginat. Uspešno smo sintetizirali tudi zamrežene encimske skupke. Encim smo oborili v zamrežene encimske skupke in z dodatkom mrežnih povezovalcev in stabilizacijskih proteinov med postopkom optimirali aktivnost encima. Primerjalno z vsemi tremi tehnikami imobilizacije encima ß-galaktozidaze se je kot najmanj uspešna izkazala ujetje encima v natrijevem alginat. Sledi sinteza zamreženih encimskih skupkov ter kovalentna vezava encima na aminosilanke in hitozanske magnetne nanodelce maghemita. Oba načina imobilizacije sta se izkazala kot zelo dobra izbira imobilizacije ß-galaktozidaze.

Keywords

beta-galaktozidaza;zamreženi encimski skupki;magnetni nanodelci maghemita;natrijev alginat;

Data

Language:	Slovenian
Year of publishing:	2016
Typology:	2.09 - Master's Thesis
Organization:	UM FKKT - Faculty of Chemistry and Chemical Engineering
Publisher:	[L. Križnik]
UDC:	543.645.4:544.478.3(043.2)
COBISS:	19530518
Views:	1434
Downloads:	185
Average score:	0 (0 votes)
Metadata:

Other data

Secondary language:	English
Secondary title:	Various techniques of immobilization of the enzyme ß-galaktozidaze
Secondary abstract:	The aim of this graduate thesis is the comparison of the chemical and physical methods of enzyme immobilisation as well as enzyme immobilisation without a carrier, e.g. to the so-called cross-linked enzyme aggregates. The β-galactosidase enzyme, which decomposes the lactose disaccharide to two monosaccharides, the glucose and galactose, was used for immobilisation. Within the graduate thesis, we have successfully synthesized maghemite magnetic nanoparticles, which have been functionalized with amino-organosilanes and hytozan. The enzyme has been covalently bonded to aminosilane and hytozan maghemite magnetic nanoparticles. We have also tried applying the enzymatic capture in an insoluble gel, for which sodium alginate was used. We have also successfully synthesized cross-linked enzyme aggregates. The enzyme was precipitated into cross-linked enzyme aggregates, whereas the enzyme activity was optimized during the process by adding cross-linking agents and stabilization proteins. Of all the three β-galactosidase enzyme immobilization methods, enzyme capture in sodium alginate turned out to be comparatively the least successful. It is followed by the cross-linked enzyme aggregate synthesis and the enzyme covalent bonding to aminosilane and hytozan maghemite magnetic nanoparticles. Both immobilization methods have turned out to be a very good choice for the β-galactosidase immobilization.
Secondary keywords:	beta-galactosidase;cross-linked enzyme aggregates;maghemite magnetic nanoparticles;sodium alginate;
URN:	URN:SI:UM:
Type (COBISS):	Master's thesis/paper
Thesis comment:	Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Pages:	IX, 63 str.
ID:	9129474