Kinetički model enzimske kaskadne reakcije za sintezu ATP-a iz adenozina

master's thesis

• Title (english): Kinetic model of an enzymatic cascade reaction for the synthesis of ATP from adenosine
• Author: Anamarija Volf
• Mentor: Martina Sudar (mentor)
• Committee member: Martina Sudar (predsjednik povjerenstva)
• Committee member: Zvjezdana Findrik Blažević (član povjerenstva)
• Committee member: Marija Vuković Domanovac (član povjerenstva)
• Granter: University of Zagreb; Faculty of Chemical Engineering and Technology; Zagreb
• Defense date and country: 2019-09-24, Croatia
• Scientific / art field, discipline and subdiscipline: TECHNICAL SCIENCES; Chemical Engineering

Abstract

U ovom je radu istražena kaskadna reakcija za sintezu adenozin trifosfata (ATP) iz adenozina. Navedenu kaskadnu reakciju čine tri reakcije pri čemu svaku od njih katalizira drugi enzim. Supstrati prve od triju proučavanih reakcija su adenozin i ATP, produkti su adenozin monofosfat (AMP) i adenozin difosfat (ADP), a reakciju katalizira enzim adenozin kinaza (ADK). Druga je reakcija između AMP-a i polifosfata (polyP) gdje kao produkt nastaje ADP, a reakciju katalizira enzim polifosfat kinaza PPK2-II. Posljednja proučavana reakcija je ona između ADP-a i polyP gdje nastaje ATP, a enzim polifosfat kinaza PPK2-I je katalizator. Svaki od triju enzima je aktivan uz odgovarajući kofaktor, Mg2+. Ispitana je kinetika sve tri pojedinačne reakcije kaskade za sintezu ATP-a metodom početnih brzina. Na temelju dobivenih rezultata procijenjeni su kinetički parametri Michaelis-Menteničinog modela te je razvijen matematički model koji se sastoji od kinetičkih jednadžbi i jednadžbi bilance tvari. Provedeni su eksperimenti u kotlastom reaktoru za svaku pojedinačnu reakciju u svrhu validacije razvijenog matematičkog modela. Primijećeno je kako tijekom dulje uporabe enzima PPK2-I i PPK2-II dolazi do pada njihove operacijske stabilnosti, odnosno do deaktivacije, što je također uvršteno u matematički model.

Abstract (english)

A cascade reaction for synthesis of adenosine triphosphate (ATP) from adenosine was studied in this work. Substrates of the first reaction are adenosine and ATP, the products are adenosine monophosphate (AMP) and adenosine diphosphate (ADP) and the enzyme that catalyzes this reaction is adenosine kinase (ADK). The second reaction is the one between AMP and polyphosphate (polyP) where the ADP is produced and reaction is catalyzed by enzyme polyphosphate kinase PPK2-II. The last of the studied reactions is the one between ADP and polyP. The product of that reaction is ATP and the reaction is catalyzed by enzyme polyphosphate kinase PPK2-I. All three enzymes are active in the presence of cofactor, Mg2+. The kinetics of the cascade reaction was investigated by the initial rate method. Based on the obtained results, the kinetic parameters of Michaelis-Menten model were estimated and the mathematical model which contains kinetic and mass balance equations was developed. Experiments in batch reactor were carried out to validate the developed mathematical model. It was noticed that during longer reaction time the activity of enzymes PPK2-I and PPK2-II decreases. In other words, these two enzymes start to deactivate, which was also included in the mathematical model.

• Keywords: kaskadna reakcija; enzim; kinetika; matematički model; kotlasti reaktor
• Keywords (english): cascade reaction; enzyme; kinetics; mathematical model; batch reactor
• Language: croatian
• URN:NBN: urn:nbn:hr:149:828027
• Study programme: Title: Applied Chemistry - Graduate 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: graduate; Academic / professional title: magistar/magistra primijenjene kemije (magistar/magistra primijenjene kemije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Closed access
• Created on: 2022-02-27 18:57:52