Operacijska stabilnost YfaU – aldolaza

undergraduate thesis

• Title (english): Operational stability of YfaU aldolases
• Author: Nikolina Jušić
• Mentor: Zvjezdana Findrik Blažević (mentor)
• Committee member: Zvjezdana Findrik Blažević (predsjednik povjerenstva)
• Committee member: Marija Vuković Domanovac (član povjerenstva)
• Committee member: Dragana Vuk (član povjerenstva)
• Granter: University of Zagreb; Faculty of Chemical Engineering and Technology; Zagreb
• Defense date and country: 2021-09-07, Croatia
• Scientific / art field, discipline and subdiscipline: TECHNICAL SCIENCES; Chemical Engineering

Abstract

U ovom radu proučavana je aldolna reakcija između piruvata i formaldehida katalizirana s tri enzima: YfaU WT, YfaU 3 i YfaU 5 - aldolazom. Radi se o tri verzije enzima od koje je prva ona prirodna, a preostale dvije su dobivene proteinskim inženjerstvom s ciljem poboljšanja svojstva stabilnosti. Kao produkt nastaje aldol natrijev 4-hidroksi-2-oksobutanoat. Natrijev piruvat je donorski supstrat, a formaldehid akceptorski supstrat. Eksperimenti su provedeni u kotlastom reaktoru te su praćeni tijek reakcije i operacijska stabilnost tri enzima. Na temelju dobivenih eksperimentalnih rezultata promjene aktivnosti enzima tijekom vremena, procijenjene su konstante deaktivacije enzima u reaktoru, pomoću programskog paketa Scientist. Osim različite brzine deaktivacije, rezultati pokazuju i različito napredovanje reakcije u reaktoru. Iz rezultata se može zaključiti da se tijekom vremena kod sva tri enzima smanjuje aktivnost, odnosno dolazi do pada operacijske stabilnosti. Najstabilnija aldolaza u kotlastom reaktoru pokazala se YfaU 5, a najmanje stabilna YfaU WT iz čega se može zaključiti da je proteinskim inženjerstvom došlo do poboljšanja stabilnosti enzima.

Abstract (english)

The aldol reaction between pyruvate and formaldehyde catalyzed by three aldolases, i.e., YfaU WT, YfaU 3 and YfaU 5, was carried out in this work. These are three versions of the enzyme, the first of which is natural, and the remaining two were obtained by protein engineering with the aim of improving the stability properties. As a product, aldol sodium 4-hydroxy-2-oxobutanoate was formed. Sodium pyruvate was a donor substrate and formaldehyde was an acceptor substrate. The experiments were carried out in a batch reactor. The course of the reaction and the operational stability of the three enzymes were monitored during the process. Based on the experimental results, i.e., the change in enzyme activity over time, the enzyme deactivation constants were estimated using the software package Scientist. In addition to different deactivation rates, the results also show different reaction progressions in the reactor. It can be concluded from the results that the activity and operational stability of all three enzymes decreases over time. The most stable aldolase in the batch reactor was YfaU 5, and the least stable was YfaU WT. Thus, it can be concluded that protein engineering led to an improvement of the enzyme stability.

• Keywords: operacijska stabilnost; YfaU - aldolaze; natrijev piruvat; formaldehid; aldolna reakcija
• Keywords (english): operational stability; YfaU - aldolase; sodium pyruvate; formaldehyde; aldol reaction
• Language: croatian
• URN:NBN: urn:nbn:hr:149:390010
• Project: Number: 635595; Title: Sustainable industrial processes based on a C-C bond-forming enzyme platform; Acronym: CARBAZYMES; Leader: Zvjezdana Findrik Blažević; Jurisdiction: eu; Funder: EC; Funding stream: H2020
• Study programme: Title: Applied Chemistry - Undergraduate study; specializations in: Environmental chemistry and green technologies, Advanced materials and technologies, Applied organic chemistry; Course: Environmental chemistry and green technologies; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) primijenjene kemije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) primijenjene kemije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2023-04-04 07:34:55