magistrsko delo
Povzetek
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 %.
Ključne besede
biokataliza;encimi;amin transaminaza;imobilizacija encimov;mikropretočni sistemi;mikroreaktorji;zamreženi encimski agregati;CLEA delci;magistrska dela;
Podatki
Jezik: |
Slovenski jezik |
Leto izida: |
2022 |
Tipologija: |
2.09 - Magistrsko delo |
Organizacija: |
UL FKKT - Fakulteta za kemijo in kemijsko tehnologijo |
Založnik: |
[Ž. Lavrič] |
UDK: |
66.098(043.2) |
COBISS: |
102137347
|
Št. ogledov: |
283 |
Št. prenosov: |
15 |
Ocena: |
0 (0 glasov) |
Metapodatki: |
|
Ostali podatki
Sekundarni jezik: |
Angleški jezik |
Sekundarni naslov: |
Preparation and application of cross-linked enzyme aggregates in microfluidic systems |
Sekundarni povzetek: |
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.
|
Sekundarne ključne besede: |
biocatalysis;enzyme immobilization;microflow systems;amin transaminase;cross-linked enzyme aggregates;Univerzitetna in visokošolska dela; |
Vrsta dela (COBISS): |
Magistrsko delo/naloga |
Študijski program: |
1000376 |
Konec prepovedi (OpenAIRE): |
2023-03-17 |
Komentar na gradivo: |
Univ. v Ljubljani, Fak. za kemijo in kemijsko tehnologijo, smer Kemijsko inženirstvo |
Strani: |
XII, 42 str. |
ID: |
14808603 |