Pročišćavanje sirovog ekstrakta enzima endo-1,4-ksilanaze upotrebom eutektičnih otapala

undergraduate thesis

• Title (english): Purification of raw endo-1,4-xylanase extract using deep eutectic solvents
• Author: Sara Anđelović
• Mentor: Bruno Zelić (mentor)
• Committee member: Bruno Zelić (predsjednik povjerenstva)
• Committee member: Anita Šalić (član povjerenstva)
• Committee member: Marko Rogošić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Chemical Engineering and Technology; Zagreb
• Defense date and country: 2022-09-12, Croatia
• Scientific / art field, discipline and subdiscipline: TECHNICAL SCIENCES; Chemical Engineering

Abstract

Ksilanaza spada u skupinu enzima ključnih za depolimerizaciju te se koristi za razgradnju linearnog polisaharida ksilana u ksilozu, razgrađujući pri tome hemicelulozu, jednu od glavnih komponenata stanične stijenke. Ksilanaza je svoju primjenu pronašla u industriji celuloze i papira, prehrambenoj industriji te kemijskoj i farmaceutskoj industriji. Kako bi se ksilanaza što učinkovitije upotrijebila, potrebno je proizvesti enzim velike katalitičke aktivnosti pri čemu je ključan korak pročišćavanje enzima. Proces pročišćavanja enzima je složen i povezan s brojnim izazovima kao što su pronalazak brzih, učinkovitih i ekološki prihvatljivih metoda uz istovremeno smanjenje proizvodnih troškova. Ekstrakcija eutektičkim otapalima (engl. Deep Eutectic Solvents, DESs) pokazala se kao učinkovita i održiva metoda ekstrakcije glicerola iz biodizela, ekstrakcije biološki aktivnih sastojaka iz prirodnih izvora, kao učinkovit reakcijski medij koji osigurava poboljšanje kontakta između reaktanata što rezultira poboljšanjem organskih sinteza i katalize. Kako bi se proces pročišćavanja enzima dodatno pospješio, kao alternativa klasičnim procesima koji se provode u šaržnim makroesktraktorima, sve više se koriste mikroekstraktori. U ovom radu provedeno je pročišćavanje sirovog ekstrakta enzima endo-1,4-ksilanaze podrijetlom iz Trametes versicolor dobivenog fermentacijom na čvrstim nosačima repinih rezanaca. Pročišćavanje enzima ksilanaza provedeno je vodenim dvofaznim sustavom kapljevina-kapljevina na bazi DES-a u šaržnom ekstraktoru i u mikroekstraktoru.

Abstract (english)

Xylanase belongs to a group of enzymes crucial for depolymerization and it is used to break down the linear polysaccharide xylan into xylose, breaking down hemicellulose, one of the main components of cell wall. Xylanase has found its application in the pulp and paper industry, the food industry and the chemical and pharmaceutical industries. In order to use xylanase as efficiently as possible, it is necessary to produce an enzyme of high catalytic activity, the key step being the purification of the enzyme. The enzyme purification process is quite complex and presents certain challenges such as finding fast, efficient and environmentally friendly methods while reducing costs. Extraction with eutectic solvents (DESs) has proven to be an effective and sustainable method of extracting glycerol from biodiesel, extracting biologically active ingredients from natural sources, as an effective reaction medium that ensures improved contact between reactants in organic synthesis and catalysis. In order to further improve the enzyme purification process, as an alternative to classical processes carried out in batch macroextractors, microextractors are being used. In this work, purification of the crude extract of endo-1,4-xylanase enzyme from Trametes versicolor produced by solid-state fermentation on beet pulp was carried out. Purification od the xylanase enzyme was performed with DES based aqueous two-phase liquid-liquid system in a batch extractor and in a microextractor.

• Keywords: enzim ksilanaza; DES; ekstrakcija; šaržni ekstratkor; mikroekstraktor
• Keywords (english): xylanase; DES; extraction; batch extractor; microextractor
• Language: croatian
• URN:NBN: urn:nbn:hr:149:403503
• Project: Number: KK.01.1.1.04.0107; Title: Biokonverzija lignoceluloznog materijala u visokovrijednu hranu za životinje; Title: Bioconversion of lignocellulosic material into high-value animal feed; Acronym: Bio4Feed; Leader: Mirela Planinić; Jurisdiction: eu; Funder: EK; Funding stream: EFRR
• Study programme: Title: Chemical Engineering - Undergraduate study; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/ prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka kemijskog inženjerstva (sveučilišni/a prvostupnik/ prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka kemijskog inženjerstva)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2023-02-17 13:15:08