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Imobilizacija encimov na magnetne mikrodelce za izvedbo kontinuirne transaminacije v mikroreaktorju z uporabo evtektičnih topil
ID Tacer, Luka (Author), ID Žnidaršič Plazl, Polona (Mentor) More about this mentor... This link opens in a new window

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Abstract
Encimi so biokatalizatorji, ki nam ponujajo mnoge prednosti pred tradicionalnimi kemijskimi katalizatorji. Omogočajo sintezo pri blagih pogojih, prav tako pa imajo izvrstno stereo-, regio- in enantioselektivnost, kar nam omogoča pridobivanje zelo čistih kompleksnih produktov z visokimi izkoristki. Kljub temu je zaradi nestabilnosti encimov pri ostrih industrijskih pogojih za njihovo vpeljavo ključna ustrezna imobilizacija. Na ta način encimom povečamo njihovo stabilnost, obenem pa omogočimo daljšo uporabo v kontinuirnih procesih. Ena izmed obetavnih tehnik imobilizacije je vezava encimov preko različnih funkcionalnih skupin na magnetne materiale, ki jih s pomočjo zunanjega magnetnega polja zadržimo v reaktorjih. Pri slednjih je v zadnjih letih ključna novost razvoj mikroreaktorske tehnologije, ki omogoča izvrsten nadzor reakcijskih pogojev, izboljšan prenos toplote in snovi ter manj stranskih produktov ter odpadkov v primerjavi s konvencionalnimi tehnologijami. V želji po zmanjšanju negativnega vpliva industrijske proizvodnje kemikalij in farmacevtskih učinkovin na okolje se uveljavlja uporaba evtektičnih topil, ki jih uvrščamo med cenovno ugodna zelena topila. Predstavljajo okolju prijazno alternativo organskim topilom, zaradi velikega nabora možnih komponent pa omogočajo mnogo fleksibilnosti pri želenih lastnostih topila. V okviru raziskovalnega dela smo pripravili več evtektičnih topil in jih ovrednotili glede na topnost benzil acetona, ki je substrat izbrane transaminacije, ter glede na zadržano aktivnost izbrane amin transaminaze ATA-v1 v topilu. Optimizirali smo postopek imobilizacije encima na magnetne mikrodelce, ki smo jih predhodno funkcionalizirali z glutaraldehidom. V izbranem evtektičnem topilu, kjer smo v ustreznem razmerju zmešali holin klorid, ureo in fosfatni pufer, smo izvedli reakcijo v mikropretočnem sistemu. Na ta način smo razvili trajnostno naravnan in učinkovit način pridobivanja metilfenilpropilamina, ki je industrijsko zanimiv kiralni produkt.

Language:Slovenian
Keywords:amin transaminaza, imobilizacija, mikropretočni sistemi, evtektična topila, magnetni mikrodelci
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:FKKT - Faculty of Chemistry and Chemical Technology
Year:2023
PID:20.500.12556/RUL-150883 This link opens in a new window
COBISS.SI-ID:173192963 This link opens in a new window
Publication date in RUL:25.09.2023
Views:672
Downloads:97
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Secondary language

Language:English
Title:Enzyme immobilisation on magnetic microparticles for continuous transamination in a microreactor using deep eutectic solvents
Abstract:
Enzymes are biocatalysts that offer many advantages compared to traditional chemical catalysts. They enable synthesis at mild reaction conditions and offer great stereo-, regio- and enantioselectivity, which leads to efficient production of highly pure complex products. However, due to enzyme instability at harsh industrial conditions, their immobilisation is crucial for their implementation at industrial scale. This way we increase enzyme stability and at the same time enable longer use of enzymes in continuous processes. One of the promising immobilisation techniques is enzyme binding on magnetic materials via different functional groups. External magnetic field is then used to hold magnetic materials in the reactor. As for the latter, microreactor technology is the most important innovation in recent years. It offers excellent control of reaction conditions, better heat and mass transfer as well as less waste and side products compared to traditional technologies. To reduce the negative environmental impact of industrial production of chemicals and pharmaceuticals, deep eutectic solvents (DESs), classified as affordable green solvents, are increasingly used. They offer an environmentally friendly alternative to organic solvents. Because of a broad range of possible components, DESs are very flexible in achieving preferred solvent properties. In this research, different DESs were prepared and ranked based on their ability to solubilize substrate of the selected transamination, namely benzyl acetone, and regarding the activity of the amine transaminase enzyme ATA-v1. The immobilisation of enzyme on magnetic microparticles that were functionalized with glutaraldehyde was optimized. The biocatalyzed transamination reaction was performed in a selected DES composed of choline chloride, urea and phosphate buffer using microflow system. Thereby a sustainable and efficient production of 1-methyl-3-phenylpropylamine, an industrialy attractive chiral amine, was developed.

Keywords:amine transaminase, immobilisation, microflow systems, deep eutectic solvents, magnetic microparticles

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