Različne tehnike imobilizacije encima beta-galaktozidaze

magistrsko delo

Lucija Križnik (Avtor), Maja Leitgeb (Mentor), Mateja Primožič (Komentor)

Povzetek

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.

Ključne besede

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

Podatki

Jezik:	Slovenski jezik
Leto izida:	2016
Tipologija:	2.09 - Magistrsko delo
Organizacija:	UM FKKT - Fakulteta za kemijo in kemijsko tehnologijo
Založnik:	[L. Križnik]
UDK:	543.645.4:544.478.3(043.2)
COBISS:	19530518
Št. ogledov:	1434
Št. prenosov:	185
Ocena:	0 (0 glasov)
Metapodatki:

Ostali podatki

Sekundarni jezik:	Angleški jezik
Sekundarni naslov:	Various techniques of immobilization of the enzyme ß-galaktozidaze
Sekundarni povzetek:	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.
Sekundarne ključne besede:	beta-galactosidase;cross-linked enzyme aggregates;maghemite magnetic nanoparticles;sodium alginate;
URN:	URN:SI:UM:
Vrsta dela (COBISS):	Magistrsko delo/naloga
Komentar na gradivo:	Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Strani:	IX, 63 str.
ID:	9129474