magistrsko delo
Abstract
Mikroreaktorji, ki so po definiciji reaktorji z vsaj eno dimenzijo pod 1mm, so vedno bolj
zanimivi industriji in vedno bolj uveljavljajo svojo pomembnost v biokatalitskih procesih.
Njihove lastnosti, kot so dobro razmerje med površino in volumnom, lahka kontrola nad
procesom, majhna poraba reagentov, omogočajo mikroreaktorjem učinkovite procese. Za
dodatno povečanje razmerja med površino in volumnom v mikroreaktorjih, je v porastu
integracija nanostruktur kot so nanovzmeti in nanotkanine, v mikroreaktorje. Omenjene
nanostrukture so se v preteklosti izkazale kot izjemen nosilec za imobilizacijo encimov z
označevalci. V okviru magistrskega dela sem razvila mikroreaktor v katerem sem želela
izvajati imobilizacijo encimov in izvedbo biotransformacije. Opravila sem
karakterizacijo nanovzmeti za določevanje hidrofobnosti. S ploščicami, ki so vsebovale
različno gosto porazdeljene nanovzmeti, sem ugotovila, da z višjo gostoto porazdelitve
nanovzmeti narašča tudi začetna hidrofobnost. V okviru karakterizacije sem opravila
analizo z vrstičnim elektronskim mikroskopom (SEM). Ugotovila sem, da je površina
nanovzmeti zgoščena in zlepljena. Predvidevam, da je to zaradi izpostavitve visoki
temperaturi med njihovo pripravo. To je tudi lahko razlog za slabo učinkovitost
nanovzmeti. ω-transaminazo z Zbasic2 označevalcem (N-SBM-ATA-wt) sem
imobilizirala na silicijeve nanovzmeti preko ionskih interakcij med peptidnim
označevalcem in nabito površino. Imobilizacija N-SBM-ATA-wt ni bila učinkovita, kar
lahko pripišem zlepljenosti in zgoščenosti nanovzmeti in njihovi neustrezni hidrofilnosti.
Karboksilno esterazo pridobljeno iz Archaeoglobus fulgidus (AF-Est2) s polihistidinskim
označevalcem sem imobilizirala na nanotkanino z bakrovimi ioni na osnovi afinitetne
imobilizacije med označevalcem in površino Ugotovila sem, da je bila imobilizacija AFEst2 na nanotkanine učinkovita, saj se je imobiliziralo tudi 100% encima, prav tako pa je
bila uspešna tudi regeneracija nanotkanine z vnosom bakrovih ionov.
Keywords
biokemijsko inženirstvo;encimi;transaminaze;esteraze;imobilizacija encimov;encimska aktivnost;mikroreaktorji;nanostrukture;biotransformacije;magistrska dela;
Data
Language: |
Slovenian |
Year of publishing: |
2020 |
Typology: |
2.09 - Master's Thesis |
Organization: |
UL FKKT - Faculty of Chemistry and Chemical Technology |
Publisher: |
[V. Erklavec Zajec] |
UDC: |
66.098:577.15(043.2) |
COBISS: |
18578947
|
Views: |
788 |
Downloads: |
189 |
Average score: |
0 (0 votes) |
Metadata: |
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Other data
Secondary language: |
English |
Secondary title: |
Enzyme immobilization on nanomaterials integrated in microreactors |
Secondary abstract: |
Microreactors, which are by definition reactors with at least one dimension below 1 mm,
are gaining more attention from industry and are more important in the field of
biocatalysis. Their favorable characteristic, such as high surface-to-volume ration, easy
process control, low reagent consumption, enable microreactors to perform highly
efficient biocatalytic processes. For additional increase of surface-to-volume ratio
nanomaterials, such as nanosprings and nanofibers, are becoming increasingly important.
Such nanostructures are an efficient carrier for tagged enzyme immobilization. In this
work a microreactor with immobilized enzymes for a desired biotransformation was
developed. Characterization of nanosprings was performed. Nanosprings with different
density were tested and it was concluded that nanosprings with higher density are more
hydrophobic at the start. An analysis with scanning electron microscope (SEM) was
performed as well. Study showed that the surface of nanosprings was melted and
concentrated. It is presumed this is the consequence of the exposure to higher
temperatures in the preparation of nanosprings. ω-transaminase with Zbasic2 (N-SBMATA-wt) tag was immobilized on silicon nanosprings via ionic interaction between
Zbasic2 tag and surface. Immobilization yield was below expectation, which could be
caused by melted nanosprings and consequently their unsuitable hydrophilic
characteristic. Carboxyl esterase from Archaeoglobus fulgidus (AF-Est2) with
polyhistidine tag was immobilized on nanofiber with functionalized Cu2+ via ionic
interactions. Immobilization yield in this experiment even reached 100%. Regeneration
of nanofibers was performed with attachment of Cu2+ particles on nanofibers and it was
successful, because further immobilizations yields were high. |
Secondary keywords: |
microreactor;immobilization;nanostructures;transaminase;esterase; |
Type (COBISS): |
Master's thesis/paper |
Study programme: |
1000376 |
Embargo end date (OpenAIRE): |
1970-01-01 |
Thesis comment: |
Univ. v Ljubljani, Fak. za kemijo in kemijsko tehnologijo, smer Kemijsko inženirstvo |
Pages: |
XII f., 59 str. |
ID: |
11779598 |