diplomsko delo univerzitetnega študijskega programa I. stopnje
Abstract
Namen našega dela je bil pripraviti zamrežene encimske skupke (CLEA) in magnetne zamrežene encimske skupke (mCLEA) iz encima β-galaktozidaze. Doseči smo želeli čim višjo preostalo aktivnost encima in čim višjo učinkovitost imobilizacije. Zamreženje smo preizkušali v štirih obarjalnih reagentih (acetonu, etanolu, 1-propanolu in 2-propanolu), dodajali smo ogrodne proteine in mrežne povezovalce v različnih kombinacijah in koncentracijah. Eksperimentalno delo je obsegalo pripravo vzorcev (obarjanje in zamreženje), aktivnostni test in Bradfordovo metodo za določanje koncentracije proteinov.
Proces smo optimirali pri konstantni koncentraciji encima (50 mg/ml) in konstantnem dodatku glutaraldehida (GA).
Ugotovili smo, da je optimalna količina dodanega mrežnega povezovalca GA 1.5% (v/v) in da mrežni povezovalec pentaetilenheksamin (PEHA) negativno vpliva na zamreženje, razen če ga dodajamo v kombinaciji z ogrodnimi proteini. V tem primeru se preostala aktivnost zviša, medtem ko učinkovitost imobilizacije močno upade.
Keywords
biokataliza;beta-galaktozidaza;imobilizacija encimov;zamreženi encimski skupki;magnetni nanodelci;diplomske naloge;
Data
Language: |
Slovenian |
Year of publishing: |
2016 |
Typology: |
2.11 - Undergraduate Thesis |
Organization: |
UM FKKT - Faculty of Chemistry and Chemical Engineering |
Publisher: |
[N. Šimenko] |
UDC: |
544.473:577.15(043.2) |
COBISS: |
20442646
|
Views: |
1224 |
Downloads: |
128 |
Average score: |
0 (0 votes) |
Metadata: |
|
Other data
Secondary language: |
English |
Secondary title: |
IMMOBILIZATION OF BETA-GALACTOSIDASE ON MAGNETIC MAGHEMITE NANOPARTICLES AND THEIR CROSS LINKING INTO ENZYME AGGREGATES |
Secondary abstract: |
The purpose of our work was to create cross-linked enzyme aggregates (CLEA) and magnetic cross-linked enzyme aggregates (mCLEA) off enzyme β-galactosidase. The aim of our work was to achive the highest possible residual activity and maximize the efficiency of immobilization. The cross-linking was tested in four most optimal precipitation reagents (acetone, ethanol, 1-propanol, 2-propanol) while stability proteins and cross-linkers in various combinations and concentrations were added to the synthesis reaction. The experimental work included preparation of samples (precipitation and cross-linking), activity test and the Bradford protein assay for determination of proteins.
The process was optimized at a constant concentration enzyme (50 mg/ml), and a constant addition of glutaraldehyde (GA).
It was concluded that the optimal quantity of added cross-linker is 1.5% (v/v) and that the cross-linker pentaetilenheksamin (PEHA) negatively affects the cross-linking, unless it is added in combination with stability proteins. In this case, the residual activity is increased, while the efficacy of immobilization drops significantly. |
Secondary keywords: |
biocatalysis;beta-galactosidase;enzyme immobilization;cross-linked enzyme aggregates;magnetic nanoparticles; |
URN: |
URN:SI:UM: |
Type (COBISS): |
Bachelor thesis/paper |
Thesis comment: |
Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo |
Pages: |
XII, 40 str. |
ID: |
9587773 |