Ekspresija rekombinantne ksiloza reduktaze na površini stanice kvasca S. cerevisiae

undergraduate thesis

• Title (english): Expression of the recombinant xylose reductase on the surface of yeast cell S. cerevisiae
• Author: Danijel Švec
• Mentor: Renata Teparić (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Chemistry and Biochemistry); (Laboratory for Biochemistry); Zagreb
• Defense date and country: 2017-09-18, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Saccharomyces cerevisiae jedan je od najčešće korištenih mikroorganizama u biotehnološkoj proizvodnji, pa se tako koristi i u proizvodnji ksilitola, prirodnog zaslađivača s velikim prehrambenim prednostima. Enzim odgovoran za mikrobiološku proizvodnju ksilitola, ksiloza reduktaza, katalizira reakciju redukcije ksiloze u ksilitol. Ksiloza reduktaza prirodno je lokalizirana u citoplazmi stanice kvasca što znači da izolacija finalnog proizvoda uključuje razbijanje stanice i više koraka pročišćavanja. Zbog relativno malog prinosa i kompleksnog pročišćavanja ksilitol na tržištu ima visoku cijenu. Tema ovog rada jest ekspresija i imobilizacija heterologne rekombinantne ksiloza reduktaze na površini stanice kvasca čime bi se izolacija proizvoda olakšala, a cijena konačnog proizvoda snizila. U tu je svrhu iskorišten protein Pir4, protein stanične stijenke kvasca, s kojim je ksiloza reduktaza fuzionirana i preko kojeg je vezana na staničnu stijenku. U radu je ispitana uspješnost ekspresije i ugradnje u stijenku te aktivnost tako imobiliziranog rekombinantnog enzima.

Abstract (english)

Saccharomyces cerevisiae is one of the most used microorganisms in biotechnological production, so it's also used in production of xylitol, natural sweetener with many nutritional advantages. Enzyme responsible for microbiological production of xylitol, xylose reductase, catalyses the reduction reaction of xylose to xylitol. Xylose reductase is naturally localized in yeast cell cytoplasm meaning that the isolation of the final product includes breaking the yeast cell and several steps of purification. Because of low yield and complexity of purification, xylitol has high price on the market. Theme of this thesis is expression and immobilization of heterologous recombinant xylose reductase on the surface of yeast cell meaning that the isolation of product would be easier and the price lower. For that purpose yeast cell wall protein Pir4is fused with xylose reductase serving as anchor through which xylose reductase binds to the yeast cell wall. In this thesis, the efficacy of expression and incorporation of the immobilized recombinant enzyme to the cell wall is examined, as well as its activity.

• Keywords: Saccharomyces cerevisiae; ksiloza reduktaza; Pir4; rekombinantni protein
• Keywords (english): Saccharomyces cerevisiae; xylose reductase; Pir4; recombinant protein
• Language: croatian
• URN:NBN: urn:nbn:hr:159:736425
• Study programme: Title: Biotechnology; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2017-09-20 09:21:11