Pročišćavanje enzima MenD za primjenu u biokatalizi

undergraduate thesis

• Title (english): Purification of enzyme MenD for application in biocatalysis
• Author: Nolla Todorović
• Mentor: Martina Sudar (mentor)
• Committee member: Anita Šalić (predsjednik povjerenstva)
• Committee member: Dajana Kučić Grgić (član povjerenstva)
• Committee member: Ana Vrsalović Presečki (član povjerenstva)
• Granter: University of Zagreb; Faculty of Chemical Engineering and Technology; Zagreb
• Defense date and country: 2021-09-23, Croatia
• Scientific / art field, discipline and subdiscipline: TECHNICAL SCIENCES; Chemical Engineering

Abstract

Razvojem biotehnologije dolazi do novih otkrića u području biokatalize čija je najveća prednost nastajanje manje štetnih nusprodukata u odnosu na kemijsku katalizu. U biokatalizi se koriste različiti oblici biokatalizatora: pročišćeni i nepročišćeni enzimi kao i cijele stanice mikroorganizma. U ovom radu proveden je postupak pročišćavanja enzima MenD (sintaza 2-sukcinil-5-enolpiruvil-6-hidroksi-3-cikloheksen-1-karboksilne kiseline) s manjom (50 mg) i većom masom uzorka (200 mg). U svrhu pročišćavanja enzima korištena je metoda afinitetne kromatografije. Nakon pročišćavanja uzoraka afinitetnom kromatografijom korištena je
SDS-PAGE elektroforeza za provjeru prisutnosti proteina u sakupljenim frakcijama te Bradfordova metoda za određivanje koncentracija proteina u frakcijama.

Abstract (english)

With the development of biotechnology, new discoveries are being made in the field of biocatalysis, the biggest advantage of which is the formation of less harmful by-products compared to chemical catalysis. Different forms of biocatalysts can be used: purified and crude enzyme as well as whole cells of the microorganisms. In this work, purification of the enzyme MenD (2-succinyl-5-enolpyruvil-6-hydroxy-3-cyclohexene-1-carboxylic acid synthase) with a lower mass (50 mg) and a higher mass sample (200 mg) was performed. Affinity chromatography was used to purify the sample. After purification of the samples by affinity chromatography, SDS-PAGE electrophoresis was used to prove the presence of proteins in the collected fractions and the Bradford method to determine protein concentration in the samples.

• Keywords: biokataliza; pročišćavanje enzima; MenD; afinitetna kromatografija; elektroforeza
• Keywords (english): biocatalysis; enzyme purification; MenD; affinity chromatography; electrophoresis
• Language: croatian
• URN:NBN: urn:nbn:hr:149:655843
• 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: Open access
• Created on: 2023-04-24 12:11:36