magistrsko delo
Žan Lavrič (Author), Polona Žnidaršič Plazl (Mentor), Marjan Marinšek (Thesis defence commission member), Igor Plazl (Thesis defence commission member)

Abstract

Imobilizacija encimov je eden najpogostejših načinov za izboljšanje stabilnosti biokatalizatorjev, ki med drugim omogoča ponovno uporabo teh učinkovitih in okolju prijaznih biokatalizatorjev. V zadnjih dveh desetletjih se je priprava zamreženih encimskih agregatov (angl. cross-linked enzyme aggregates, CLEA) izkazala kot zelo učinkovit, enostaven in cenovno ugoden način imobilizacije. V magistrski nalogi smo razvili postopek priprave zamreženih encimskih agregatov izbrane amin transaminaze (ATA-CLEA) v mikropretočnem sistemu. Povprečen radij delcev je bil 37,13 ± 0,38 nm, kar je bistveno manj kot pri šaržnem postopku, kjer nastanejo delci opazni prostemu očesu. ATA-CLEA delci, pripravljeni v mikropretočni napravi, so izkazali 87,1 % zadržane aktivnosti, kar je po literaturnih podatkih kar 2,4-krat več kot pri CLEA delcih, pridobljenih v šaržnem postopku Poleg tega smo v mikropretočnem sistemu razvili in-situ ločevanje agregatov od raztopine, s čimer smo se izgonili običajnim postopkom, ki v večini primerov vodijo v zlepljenje nastalih delcev in manjšo zadržano aktivnost pripravkov. Namesto centrifugiranja smo nastale delce v mikropretočnem sistemu vodili v pretočni membranski mikroreaktor, v katerem smo lahko enostavno odstranili reagente za njihovo tvorbo, ob enem pa s tem encimski pripravek imobilizirali za nadaljnjo biotransformacijo. Proces z ATA-CLEA delci v membranskem mikroreaktorju je bil bolj učinkovit v primerjavi z neagregiranim encimom. ATA-CLEA delci so tudi bolj stabilni od neagregiranega encima, saj je bila produktivnost membranskega mikroreaktorja z ATA-CLEA delci v šestih dneh kontinuirnega procesa nespremenjena, medtem ko je pri neagregiranem encimu produktivnost v petih dneh padla za 20 %.

Keywords

biokataliza;encimi;amin transaminaza;imobilizacija encimov;mikropretočni sistemi;mikroreaktorji;zamreženi encimski agregati;CLEA delci;magistrska dela;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UL FKKT - Faculty of Chemistry and Chemical Technology
Publisher: [Ž. Lavrič]
UDC: 66.098(043.2)
COBISS: 102137347 Link will open in a new window
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Other data

Secondary language: English
Secondary title: Preparation and application of cross-linked enzyme aggregates in microfluidic systems
Secondary abstract: Enzyme immobilization is one of the most common ways to improve the stability of biocatalyst, which, among other things, allows the reuse of environmentally friendly biocatalysts. In the last two decades, the preparation of cross-linked enzyme aggregates (CLEA) has proven to be a very effective, fast, and cost-effective way of immobilization. A novel process for the preparation of cross-linked enzyme aggregates of selected amine transaminase (ATA-CLEA) in the microflow system was developed. The mean particle radius was 37.13 ± 0.38 nm, which is less than in batch process, where particles visible to the neaked eye are formed. ATA-CLEA particles prepared in microflow process thus showed 87.1 % of retained activity, which according to literature is 2.4-times more than for CLEA particles obtained in the batch process. In addition, in-situ separation of the formed aggregates from the solution was developed and thus eliminating the usual techniques, which in most cases lead to sticking of the formed particles which lowers retained activity. Instead of centrifugation, the formed particles in the microflow system were led to a flow membrane microreactor in which reagents for their formation could be easily removed, and at the same time the enzyme preparation was immobilized for further biotransformation. The process with ATA-CLEA particles in the membrane microreactor is more efficient compared to the non-aggregated enzyme. ATA-CLEA particles are also more stable than non-aggregated enzymes, as the productivtiy of the membrane microreactor with ATA-CLEA particles was unchanged for six days of continuous process, while the non-aggregated enzyme’s productivity fell by 20 % in five days.
Secondary keywords: biocatalysis;enzyme immobilization;microflow systems;amin transaminase;cross-linked enzyme aggregates;Univerzitetna in visokošolska dela;
Type (COBISS): Master's thesis/paper
Study programme: 1000376
Embargo end date (OpenAIRE): 2023-03-17
Thesis comment: Univ. v Ljubljani, Fak. za kemijo in kemijsko tehnologijo, smer Kemijsko inženirstvo
Pages: XII, 42 str.
ID: 14808603