Ekstrakcija enzima RuBisCO iz šećerne repe pomoću visokonaponskog električnog pražnjenja

master's thesis

• Title (english): Extraction of RuBisCO enzyme from sugar beet by means of high voltage electrical discharges
• Author: Renata Mavrić
• Mentor: Anet Režek Jambrak (mentor)
• Mentor: Josipa Dukić (pomoć pri izradi, neposredni voditelj)
• Mentor: Karla Košpić (pomoć pri izradi, neposredni voditelj)
• Committee member: Anita Slavica (predsjednik povjerenstva)
• Committee member: Mojca Čakić Semenčić (član povjerenstva)
• Committee member: Igor Stuparević (član povjerenstva)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Management, Foreign Languages for Specific Purposes and Physical Education and Health); (Laboratory for Sustainable Development); Zagreb
• Defense date and country: 2023-09-25, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Food Technology

Abstract

Provedeno je istraživanje utjecaja parametara visokonaponskog električnog pražnjenja (HVED) na ekstrakciju ribuloza-1,5-bisfosfat karboksilaza/oksigenaza (RuBisCO) enzima iz svježih listova šećerne repe s ciljem valorizacije otpada šećerne industrije. HVED ekstrakcija provedena je uz uvođenje argona ili dušika pri 20 – 25 kV i 3 – 9 min. Izmjerena koncentracija ukupnih proteina u ekstraktu varirala je od 14,27 ± 0,53 mg/g s.t. do 34,90 ± 1,59 mg/g s.t., ovisno o primijenjenom naponu, vremenu tretmana i o radnome plinu. Optimalni parametri HVED ekstrakcije za maksimalni prinos ukupnih proteina su napon 20 kV i vrijeme tretmana 3 min. Western blot metodom detektirana je velika podjedinica RuBisCO enzima u području 55 kD. Kvatifikacija MRM (engl. multiple reaction monitoring) metodom pokazala je da radni plin znatno utječe na prinos koncentracije RuBisCO enzima. Prinos RuBisCO enzima HVED ekstrakcijom pri 25 kV i 6 min uz argon iznosi 81,32 ± 0,73 µg/mL, dok uz dušik 11,83 ± 0,02 µg/mL.

Abstract (english)

Research was conducted to determine the influence of high voltage electrical discharges (HVED) parameters on the extraction of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) enzyme from fresh sugar beet leaves, aiming to valorize sugar industry waste. HVED extraction was executed at voltages of 20 – 25 kV for 3 – 9 minutes while introducing Argon or Nitrogen. Measured concentration of total proteins in the extracts varied from 14.27 ± 0.53 mg/g d.m. to 34.90 ± 1.59 mg/g d.m., depending on the applied voltages, treatment time and input gas. The optimal HVED extraction parameter to obtain maximal yield was 20 kV and 6 minutes. RuBisCO enzyme's big subunit was detected at 55 kDa. Quanitification of RuBisCO enzyme using multiple reaction monitoring method showed the impact of input gas on the yield of RuBisCO. Yield of RuBisCO at 25 kV and 6 min using Argon was 81.32 ± 0.73 µg/mL, while using Nitrogen 11.83 ± 0.02 µg/mL.

• Keywords: RuBisCO; šećerna repa; plazma; HVED; netoplinske tehnike; valorizacija otpada
• Keywords (english): RuBisCO; sugar beet; plasma; HVED; nonthermal processing; waste valorization
• Language: croatian
• URN:NBN: urn:nbn:hr:159:899389
• Study programme: Title: Food Engineering; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka prehrambenog inženjerstva (magistar/magistra inženjer/inženjerka prehrambenog inženjerstva)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-07-02 09:55:22