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Imobilizacija transaminaz na prepletena nanovlakna v mikroreaktorjih
Šketa, Borut (Author), Žnidaršič Plazl, Polona (Mentor) More about this mentor... This link opens in a new window

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Abstract
Encimska kataliza omogoča izvedbo enantioselektivnih reakcij pri nizkih temperaturah, blagem pH in atmosferskem tlaku. Kljub tem prednostim pa je za ekonomsko upravičenost biokatalitskih procesov v industriji ključna imobilizacija encimov. Ta zagotovi manjše izgube dragih encimov, hkrati pa lahko izboljša njihovo stabilnost in odpornost v pogojih industrijskih procesov, ki zelo odstopajo od naravnega okolja encimov. Izvedba biokatalitskih procesov v mikropretočnih sistemih, ki zaradi dobre regulacije temperature, prenosa snovi in majhnega volumna omogočajo visoke konverzije ob majhnih količinah porabljenih topil ter nastalih stranskih produktov, hkrati pa nudijo možnost kontinuirnega delovanja, v zadnjem času zavzema vse več pozornosti. Prepletena nanovlakna predstavljajo prilagodljiv nosilec za imobilizacijo encimov v mikroreaktorje. Imajo veliko specifično površino z možnostjo funkcionalizacije, obenem pa zaradi velike poroznosti omogočajo skoraj nemoten transport snovi. V pričujočem delu smo izrazili in izolirali več amin transaminaz s heksahistidinskm (His6) označevalcem na različnih koncih proteina in brez njega ter primerjali njihovo imobilizacijo na prepletenih nanovlaknih z različnimi funkcionalnimi skupinami. Na nanovlaknih, na katere se je transaminaza vezala kovalentno, smo opazili veliko izgubo aktivnosti, medtem ko je koordinativna vezava preko His6 označevalca zadržala največ prvotne encimske aktivnosti. Poleg tega smo na izbranih prepletenih nanovlaknih uspešno izvedli koimobilizacijo kofaktorja encima.

Language:Slovenian
Keywords:imobilizacija, amin transaminaza, mikropretočni sistemi, biotransformacija
Work type:Master's thesis/paper (mb22)
Tipology:2.09 - Master's Thesis
Organization:FKKT - Faculty of Chemistry and Chemical Technology
Year:2021
COBISS.SI-ID:86441731 This link opens in a new window
Views:104
Downloads:40
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Secondary language

Language:English
Title:Transaminases immobilization on nanofiber mats in microreactors
Abstract:
Enzyme catalysis allows for enantioselective reactions to be performed at low temperatures, mild pH and atmospheric pressure. Despite these advantages, the immobilization of enzymes is crucial for the economic viability of biocatalytic processes in industry. Immobilization ensures lower losses of expensive enzymes and at the same time can improve their stability under the conditions of industrial processes that deviate greatly from the natural environment of enzymes. The implementation of biocatalytic processes in microflow systems allows for high conversions with small amounts of solvents and by-products, due to good temperature regulation, fast mass transfer and low volume. At the same time microflow systems offer the possibility of continuous operation and have received much attention lately. Nanofiber mats represent a flexible carrier for the immobilization of enzymes in microreactors. They have a large specific surface with the possibility of functionalization and at the same time, due to their high porosity, they enable efficient mass transport. In this work, expression and isolation of several amine transaminases with and without hexahistidine (His6) tags at different ends of the protein was performed and compared regarding their immobilization efficiency on nanofiber mats with different functional groups. A large loss of enzyme activity was observed when transaminases were covalently bound to the nanofiber mat, while coordinate binding via His6 tag resulted in the highest retained enzyme activity. In addition, the cofactor co immobilization on a selected nanofiber mat was successful.

Keywords:immobilization, amine transaminase, microfluidic systems, biotransformation

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