Imobilizacija ß-laktamaze

magistrsko delo

Adam Brumen (Author), Maja Leitgeb (Mentor), Mateja Primožič (Co-mentor), Katja Heržič (Co-mentor)

Abstract

Namen magistrskega dela je predstaviti pomen encima β-laktamaze za javno zdravje, kemijske in fizikalne metode imobilizacije encimov ter magnetne nanodelce kot nosilce za imobilizacijo encimov. V okviru naloge smo izvedli imobilizacijo β-laktamaze na magnetne nanodelce, ki se lahko uporablja za razgradnjo antibiotikov. β-Laktamaza razgrajuje antibiotike z β-laktamskimi obroči, natančneje peniciline, cefalosporine in karbapeneme. V okviru raziskovalnega dela smo uspešno sintetizirali aminosilanske magnetne nanodelce, ki smo jih uporabili za imobilizacijo encima. Z dodajanjem mrežnih povezovalcev in stabilizacijskih proteinov ter s spreminjanjem sinteznih parametrov, kot so temperatura, pH, vrtilna hitrost in čas imobilizacije, smo določili optimalne parametre za imobilizacijo obravnavanega encima na magnetne nanodelce. Pri raziskovalnem delu smo tako uporabili 20 mg aminosilanskih nanodelcev in encim β-laktamazo s koncentracijo 0,01 mg/mL. Ugotovili smo, da je optimalen dodatek mrežnega povezovalca glutaraldehida 10 % (v/v), optimalen dodatek govejega seruma albumina 25 % (v/v), vrtilna hitrost 450 rpm in čas imobilizacije dve uri. Proučevali smo tudi stabilnost prostega in imobiliziranega encima. Izvedli smo tudi študijo večkratne uporabe imobiliziranega encima. Po petnajstih ciklih ponovne uporabe je imobilizirani encim ohranil 12,3 % svoje začetne aktivnosti.

Keywords

imobilizacija encima;magnetni nanodelci;β-laktamaza;β-laktamski antibiotiki;mrežni povezovalec;diplomske naloge;

Data

Language:	Slovenian
Year of publishing:	2023
Typology:	2.09 - Master's Thesis
Organization:	UM FKKT - Faculty of Chemistry and Chemical Engineering
Publisher:	[A. Brumen]
UDC:	534.645.4(043.2)
COBISS:	171619331
Views:	74
Downloads:	22
Average score:	0 (0 votes)
Metadata:

Other data

Secondary language:	English
Secondary title:	Imobilization of ß-lactamase
Secondary abstract:	The purpose of the master's thesis is to present the importance of the β-lactamase enzyme to public health, chemical and physical methods of enzyme immobilization, and magnetic nanoparticles as carriers for immobilization. For immobilization, we used the enzyme β-lactamase, which degrades antibiotics with β-lactam rings, specifically penicillins, cephalosporins and carbapenems, and therefore represents one of the major clinical problems. As part of the research work, we successfully synthesized maghemite aminosilane nanoparticles, which were used for enzyme immobilization. By adding network linkers and stabilizing proteins and by changing parameters such as temperature, pH, rotation speed and immobilization time, we determined the optimal parameters for the immobilization of the enzyme in question on magnetic nanoparticles. In the research work, we used 20 mg of aminosilane nanoparticles and the enzyme β-lactamase with the conentration of 0,01 mg/mL and found that that the optimal addition of the crosslinker glutaraldehyde is 10% (v/v), the optimal addition of bovine serum albumin is 25% (v/v), the rotation speed is 450 rpm and the immobilization time is two hours. Incubation caused a steep drop in the residual activity of the free enzyme over time, while the activity of the immobilized enzyme was almost completely preserved. After fifteen cycles of use, the immobilized enzyme retained 12.3 percent of the activity compared to the immobilized enzyme at the first use and 14 percent compared to the free enzyme at the first use.
Secondary keywords:	enzyme immobilization;magnetic nanoparticles;β-lactamase;β-lactam antibiotics;cross-linking agent;
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, 41 f.))
ID:	19965307