Određivanje aktivnosti saharoza fosforilaze HPAE-PAD metodom

undergraduate thesis

• Title (english): Determination of activity of sucrose phosphorylase by HPAE-PAD method
• Author: Antonia Krsnik
• Mentor: Anita Slavica (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: 2021-07-09, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Aktivnost enzima može se uspješno popratiti određivanjem koncentracije supstrata i/ili produkata enzimske reakcije i to metodama visoko učinkovite tekućinske kromatografije. Saharoza fosforilaza (EC 2.4.1.7) se koristi u industrijskoj proizvodnji α-glukozilglicerola, koji nastaje regioselektivnom glukozilacijom glicerola. Anionsko-izmjenjivačka kromatografija visoke učinkovitosti sa pulsnom amperometrijskom detekcijom (HPAE-PAD) pokazala se kao metoda pogodna za određivanje koncentracije svih spojeva koji sudjeluju u reakciji koju katalizira saharoza foforilaza, a time i procijenu aktivnosti ovoga enzima. Stoga su u ovome završnome radu objedinjeni ključni parametri HPAE-PAD metode za određivanje koncentracije spojeva od interesa, kao i njena teorijska pozadina, čime je poduprta prikladnost HPAE-PAD metode za određivanje aktivnosti saharoza fosforilaze. Posebna pozornost je posvećena fizikalno-kemijskim svojstvima odabranih ugljikohidrata i industrijski važnih i visoko vrijednih biokemikalija, koji nastaju konverzijom ovih supstrata. Dodatno, pobliže su opisani uvjeti pri kojima bi se odabrana analitička metoda provodila i dati su očekivani rezultati za slučaj kada bi analiza ovom metodom bila provedena.

Abstract (english)

Enzyme activity can be successfully followed by determination of substrates and/or products concentration by high-pressure liquid chromatography methods. Sucrose phosphorylase (EC 2.4.1.7) is employed in industrial manufacturing of α-glucosylglycerol which is produced by regioselective glucosylation of glycerol. High pressure anion-exchange chromatography with pulsed amperometric detection (HPAE-PAD) is a suitable method for concentration determination of all compounds involved in a reaction catalized by the sucrose phospohrylase, and therefrom for estimation of the enzyme activity. Therefore, in this graduate thesis all key parameters of the HPAE-PAD method for concentration determination of all given compounds were comprised, as well as a theoretical background of the method. In this way suitability of the HPAE-PAD method for sucrose phospohrylase activity estimation was supported. Special attention was paid to physical and chemical properties of selected carbohydrates and industrially important high-added value biochemicals obtained by conversion of those substrates. In addition, analytical conditions and expected results of predicted analysis by the HPAE-PAD method were described in many details.

• Keywords: α-glukozilglicerol; HPAE-PAD metoda; saharoza fosforilaza
• Keywords (english): α-glucosylglycerol; HPAE-PAD method; sucrose phosphorylase
• Language: croatian
• URN:NBN: urn:nbn:hr:159:521159
• 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: 2021-07-23 11:42:27