Kinetička karakterizacija ketoreduktaze

undergraduate thesis

• Title (english): Kinetic characterization of the ketoreductase
• Author: Vanda Pakšec
• Mentor: Ana Vrsalović Presečki (mentor)
• Committee member: Ana Vrsalović Presečki (predsjednik povjerenstva)
• Committee member: Vanja Kosar (član povjerenstva)
• Committee member: Dajana Kučić Grgić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Chemical Engineering and Technology; Zagreb
• Defense date and country: 2019-07-12, Croatia
• Scientific / art field, discipline and subdiscipline: TECHNICAL SCIENCES; Chemical Engineering

Abstract

U ovom radu ispitivana je kinetika reakcije redukcije propanala uz biokatalizator ketoreduktazu (KRED), koji za svoju aktivnost zahtijeva prisustvo koenzima nikotin-amid-adenin-dinukleotid-fostata (NADPH). Kinetika je određena metodom početnih brzina reakcije, mjerenjem ovisnosti brzine reakcije o koncentraciji supstrata (propanal, NADPH). Također ispitao se i utjecaj produkta (propanola i NADP+) na početnu brzinu reakcije. Kinetika reakcije je opisana dvosupstratnom Michaelis-Menteničinom kinetikom s kompetitivnom inhibicijom produktima, a parametri su procijenjeni nelinearnom regresijom. Ispitivanje aktivnosti enzima u povratnoj reakciji pokazalo je da KRED ne katalizira reakciju oksidacije propanola. Provedena su mjerenja utjecaja propanala na stabilnost biokatalizatora, variranjem koncentracije propanala u reaktoru. Na temelju izmjerenih ovisnosti postavljen je matematički model enzimske reakcije, te su napravljene simulacije procesa.

Abstract (english)

In this paper, the kinetics of ketoreductase (KRED) was investigated in the reaction of propionaldehyde reduction. For its activity, this enzyme requires the use of coenzyme nicotinamide-adenine dinucleotide phosphate (NADPH). The kinetics was determined by the initial reaction rate method, by measuring dependence on reaction rate on substrate concentration (propionaldehyde and NADPH). Also, the impact of product (propanol and NADP+) on initial reaction rate was examined. Reaction kinetics was described by double substrate Michaelis–Menten kinetics with include competitive product inhibition. Kinetic parameters of Michaelis-Menten equation were estimated by nonlinear regression The enzyme did not show any measurable activity in reverse reaction, oxidation of propanol. Stability of enzyme was measured in the presence of different concentrations of propionaldehyde. Based on the experimental data mathematical model was set and the process simulations have been done.

• Keywords: enzimska kinetika; simulacija procesa; matematički model; ketoreduktaze
• Keywords (english): enzyme kinetics; process simulation; matematical modelling; ketoreductase
• Language: croatian
• URN:NBN: urn:nbn:hr:149:718130
• 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; Embargo expiration date: 2019-07-12
• Created on: 2023-05-26 10:00:43