Recombinant expression, purification and characterisation of two biomass degrading enzymes in Trichoderma reesei

master's thesis

• Title (croatian): Rekombinantna ekspresija, pročišćavanje i karakterizacija dvaju enzima za razgradnju biomase u Trichoderma reesei
• Author: Alen Požgajčić
• 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

Plijesni, kao saptrofitski mikroorganizmi, igraju glavnu ulogu u zemljinom ciklusu ugljika na način da njihova enzimatska mašinerija rastavlja biljnu biomasu na jednostavne šećere. Kao primjer dva takva enzima, litička polisahardina monooksigenaza (LPMO) i celobioza dehidrogenaza (CDH) pokazuju sinergističku aktivnost. Njihova aktivnost pokazuje visoki potencijal za saharifikaciju biomase i proizvodnju biogoriva na industrijskoj razini. U ovom radu, CDH iz plijesni Neurospora crassa je ekspresioniran u plijesni Trichoderma reesei, pročišćen i karakterizirani su mu kinetički parametri zajedno s pH i temperaturnim optimumom. Istovremeno, inhibitori LPMOa su karakterizirani kako bi se objasnili mehanizmi inhibicije u prirodi.

Abstract (english)

Fungi, as a saprophytic microorganisms, play a central role in the terrestrial carbon cycle in a way that their enzymatic machineries deconstructs plant biomass to simple sugars. Two of the enzymes from that machinery, lytic polysaccharide monooxygenase (LPMO) and cellobiose dehydrogenase (CDH) have shown synergistic activity. Their activity shows high potential for industrial scale saccharification of biomass and production of biofuels. In this work, CDH from fungus Neurospora crassa has been expressed in fungus Trichoderma reesei, purified and characterised its kinetical parameters together with pH and temperature optima. Simultaneously, characterisation of LPMO inhibitors have been done so that we can explain the mechanisms of inhibition in the nature.

• Keywords: CDH; LPMO; Trichoderma reesei; Neurospora crassa; rekombinantna ekspresija; pročišćavanje; karakterizacija; inhibicija
• Keywords (english): CDH; LPMO; Trichoderma reesei; Neurospora crassa; Recombinant expression Purification; Characterisation; Inhibition
• Language: english
• URN:NBN: urn:nbn:hr:159:710407
• 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: 2019-11-27 11:37:33