Novel laccases from Fomitopsis Pinicola in Trichoderma Reesei

master's thesis

• Title (croatian): Rekombinantna ekspresija, pročišćavanje i karakterizacija dviju novih lakaza iz Fomitopsis Pinicola u Trichoderma Reesei
• Author: Mihael Colar Zanjko
• Mentor: Božidar Šantek (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for Biochemical Engineering, Industrial Microbiology and Malting and Brewing Technology); Zagreb
• Defense date and country: 2018-11-26, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Humanity is currently facing two great challenges, namely global warming and the energy crisis. As a possible solution the use of renewable energy and biodegradable materials is emphasized (e.g. biomass). For the production of bioenergy, mainly bioethanol, from biomass in a eco-friendly way, enzymes are needed to degrade lignocellulosic materials. Despite considerable efforts have been made in their investigation, much is still uknown about these enzymes and the way they interact when degrading their natural substrates. The first step to resolve these questions is the isolation and characterisation of those enzymes. Keeping this in mind, the aim of this work was to obtain two such enzymes, namely laccases, from the fungus Fomitopsis pinicola by recombinant expression in the fungus Trichoderma reesei, and to subsequently purify and characterise them. Both laccases were purified to about 80% purity, and, among other things, their pH optima were determined, as well as kinetic constants for three different substrates at differnet pH values.

Abstract (croatian)

Čovječanstvo se u današnje doba suočava sa dva velika izazova, a to su globalno zatopljenje i kriza energenata. Kako bi se tome stalo na kraj stavlja se naglasak na obnovljivu energiju i materijale (npr. biomasa). Za dobivanje energenata poput bioetanola iz biomase na ekološki neškodljiv način potrebni su enzimi koji razgrađuju lignocelulozne sirovine. No, usprkos mnogim istraživanjima još je mnogo toga nepoznato o tim enzimima i njihovom međudjelovanju tijekom razgradnje biomase, a prvi korak ka razrješenju svih tih pitanja predstavlja izolacija i karakterizacija tih enzima. S obzirom na navedeno, svrha ovog rada bila je dobivanje dvaju takvih enzima, lakaza, iz plijesni Fomitopsis pinicola pomoću rekombinantne ekspresije u plijesni Trichoderma reesei, te njihovo pročišćavanje i karakterizacija. Obje lakaze pročišćene su do 80% čistoće, te su im, između ostalog, određeni pH optimumi koji se nalaze u niskom pH području, kao i kinetičke konstante za tri različita supstrata kod dvije različite pH vrijednosti.

• Keywords: laccase; fermentation; Fomitopsis pinicola; Trichodema reesei; recombinant expression; enzyme kinetics
• Keywords (english): lakaza; fermentacija; Fomitopsis pinicola; Trichoderma reesei; rekombinantna ekspresija; kinetika enzima
• Language: english
• URN:NBN: urn:nbn:hr:159:872547
• Study programme: Title: Bioprocess Engineering; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka bioprocesnog inženjerstva (magistar/magistra inženjer/inženjerka bioprocesnog inženjerstva)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2019-11-14 11:44:44