Pročišćavanje i karakterizacija enzima lipaza porijeklom iz Thermomyces lanuginosus

undergraduate thesis

• Title (english): Purification and characterization of lipase from Thermomyces lanuginosus
• Author: Tomislav Marčinko
• Mentor: Anita Šalić (mentor)
• Committee member: Anita Šalić (predsjednik povjerenstva)
• Committee member: Bruno Zelić (član povjerenstva)
• Committee member: Marija Vuković Domanovac (član povjerenstva)
• Granter: University of Zagreb; Faculty of Chemical Engineering and Technology; Zagreb
• Defense date and country: 2020-09-22, Croatia
• Scientific / art field, discipline and subdiscipline: TECHNICAL SCIENCES; Chemical Engineering

Abstract

Lipaze su važni izvanstanični, termostabilni enzimi koje su svoju primjenu pronašli u prehrambenoj, papirnoj, farmaceutskoj industriji te proizvodnji alkoholnih i bezalkoholnih napitaka. Jedan od načina proizvodnje lipaza je uzgoj različitih mikroorganizama (gljive, kvasci, bakterije) na čvrstim nosačima. U zadnje vrijeme, kako bi cijelu proizvodnju lipaza na čvrstim nosačima učinili ekonomičnijom, kao supstrat se koristi otpad poljoprivrednog podrijetla. Bez obzira na način proizvodnje, dobiveni enzim lipaza se mora pročistiti kako bi bio komercijalno upotrebljiv, a sam mehanizam pročišćavanja predstavlja jedan od najvećih izazova. Glavni cilj ovoga procesa je brzo i učinkovito pročistiti enzim kako bi se smanjili troškovi često vrlo skupih procesa pročišćavanja. Cilj ovog rada bio je pročistiti enzim lipazu porijeklom iz mikroorganizma Thermomyces lanuginosus dobivenog fermentacijom na čvrstim nosačima pogače buče. Kao prvi korak u procesu pročišćavanja sirovog ekstrakta enzima lipaze provedeno je isoljavanje amonijevim solima nakon čega je uslijedila dijaliza. Kako bi se enzim dodatno pročistio provedena je kromatografija isključenja po veličini na Sephadex G-100 koloni. Nakon provedbe procesa pročišćavanja, provedena je karakterizacija pročišćenog i nepročišćenog enzima lipaze na način da su određeni temperaturni i pH optimum, stabilnost te utjecaj metalnih iona, organskih otapala, masti i ulja na aktivnost enzima.

Abstract (english)

Lipases are important extracellular, thermostable enzymes that have found their application in the food, paper and pharmaceutical industries and in the production of alcoholic and non-alcoholic beverages. One of the ways of lipase production is production on solid carriers using different microorganisms (fungi, yeasts, bacteria). Recently, in order to make the whole lipase production process more economical, agricultural waste is used as a substrate. However, regardless of the production process, the resulting lipase must be purified in order to be commercially usable, and the purification mechanism itself is one of the biggest challenges. The main goal is to purify the enzyme quickly and efficiently in order to reduce the costs of often very expensive purification processes. The aim of this work was to purify the enzyme lipase from Thermomyces lanuginosus obtained by fermentation on solid carriers of pumpkin cake. As a first steps in the purification process of the crude lipase enzyme extract, salting with ammonium salts followed by dialysis were performed. Size exclusion chromatography on a Sephadex G-100 column was performed to further purify the enzyme. After the purification, the purified and crude lipase was characterized by determining the temperature and pH optimum, stability and the influence of metal ions, organic solvents, fats and oils on the activity of the enzyme.

• Keywords: Thermomyces lanuginosus; enzim lipaza; pročišćavanje; karakterizacija
• Keywords (english): Thermomyces lanuginosus; lipase; purifying; characterization
• Language: croatian
• URN:NBN: urn:nbn:hr:149:192294
• Project: Number: IP-2016-06-7993; Title: Razvoj integriranog mikrosustava za biokatalitičku proizvodnju biodizela; Acronym: DeMSy(BioPro)2; Leader: Bruno Zelić; Jurisdiction: Croatia; Funder: HRZZ; Funding stream: IP
• Study programme: Title: Environmental Engineering - Undergraduate study; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/ prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka ekoinženjerstva (sveučilišni/a prvostupnik/ prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka ekoinženjerstva)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2023-05-05 09:37:50