Proizvodnja S-adenozil-L-metionina pomoću kvasca Pichia stipitis

doctoral thesis

• Title (english): Production of S-adenosyl-L-methionine by yeast Pichia stipitis
• Author: Stela Križanović
• Mentor: Damir Stanzer (mentor)
• Committee member: Jasna Mrvčić (predsjednik povjerenstva)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; Zagreb
• Defense date and country: 2016-07-07, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology
• Universal decimal classification (UDC): 577 - Biochemistry. Molecular biology. Biophysics

Abstract

Cilj ovog rada je bio ispitati mogućnost proizvodnje S-adenozil-L-metionina (SAMe) pomoću kvasca Pichia stipitis CBS 5776 (P. stipitis). Primjenom UV-mutageneze i nistatina selekcioniran je soj M12 kvasca P. stipitis koji ne eksprimira C-24 sterol metiltransferazu (Erg6p) čime je narušena biosinteza ergosterola te povećana proizvodnja SAMe. Soju M12 kvasca P. stipitis određen je genetski i proteomski profil te optimalan sastav podloge (vrsta osnovnog izvora ugljika, količina i oblik metionina, prisutnost soli) kao i optimalni uvjeti uzgoja (aeracija, temperatura i pH-vrijednost) za proizvodnju SAMe. Uzgojem soja M12 kvasca P. stipitis na glicerolu kao osnovnom izvoru ugljika uz pulsni dodatak metionina na početku stacionarne faze rasta ostvareno je povećanje sadržaja SAMe u biomasi soja M12 kvasca P. stipitis u odnosu na divlji soj kvasca P. stipitis.

Abstract (english)

The aim of this work was to evaluate the possibility of S-adenosyl-L-methionine (SAM) production using yeast Pichia stipitis CBS 5776 (P. stipitis). P. stipitis M12 strain was selected using UV-mutagenesis and nystatin. M12 strain does not express C-24 sterol metyltransferase (Erg6p), and thus has decreased ergosterol biosynthesis and increased production of SAM. Genetic and proteomic profile for P. stipitis M12 strain, as well as optimal medium composition (a type of the main carbon source, amount and form of methionine, presence of salts) and growth conditions (aeration, temperature and pH) for SAM production were determined. An increase in SAM content in the biomass by growing M12 strain on glycerol as the main carbon source with a pulse addition of methionine at the beginning of the stationary growth phase, compared to the P. stipitis wild strain was achieved.

• Keywords: S-adenozil-L-metionin; Pichia stipitis (P. stipitis); C-24 sterol metiltransferaza (Erg6p); proteom; glicerol
• Keywords (english): S-adenosyl-L-methionine; Pichia stipitis (P. stipitis); C-24 sterol methyltransferase (Erg6p); proteome; glycerol
• Language: croatian
• URN:NBN: urn:nbn:hr:159:250535
• Study programme: Title: Postgraduate PhD Study in Biotechnology and Bioprocess Engineering; Study programme type: university; Study level: postgraduate; Academic / professional title: doktor/doktorica znanosti (doktor/doktorica znanosti)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2018-05-17 12:47:34