Aktivnosti aldehid-dehidrogenaza u reakciji oksidacije aldola

undergraduate thesis

• Title (english): Activities of aldehyde dehydrogenases in aldol product oxidation
• Author: Paula Patrčević
• Mentor: Zvjezdana Findrik Blažević (mentor)
• Committee member: Zvjezdana Findrik Blažević (predsjednik povjerenstva)
• Committee member: Martina Sudar (član povjerenstva)
• Committee member: Dajana Kučić Grgić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Chemical Engineering and Technology; Zagreb
• Defense date and country: 2019-09-19, Croatia
• Scientific / art field, discipline and subdiscipline: TECHNICAL SCIENCES; Chemical Engineering

Abstract

U ovom radu provedena je reakcija oksidacije 3-hidroksi-2-metilpropanala u odgovarajuću 3-hidroksi-2-metilpropansku kiselinu. Glavni cilj rada bila je potraga za odgovarajućom aldehid-dehidrogenazom koja najuspješnije može oksidirati produkt aldolne adicije, iz zadanog panela od 25 aldehid-dehidrogenaza. Kako produkt aldolne adicije (ujedno supstrat - 3-hidroksi-2-metilpropanal) nije komercijalno dostupan, bilo ga je potrebno sintetizirati prije provedbe reakcije oksidacije pomoću enzima D-fruktoze-6-fosfat aldolaze (FSA D6Q). Reakcija oksidacije aldola u odgovarajuću karboksilnu kiselinu provodi se pomoću dvaju enzima (aldehid dehidrogenaza i NADH oksidaze) te koenzima NAD+. NADH oksidaza (NOX) je enzim koji prevodi NADH u NAD+, a koristi se za regeneraciju koenzima NAD+ kako bi njegova potrošnja u reaktoru bila minimalna. Aldehid-dehidrogenaza 41 pokazala se kao najuspješniji biokatalizator za istraživanu reakciju oksidacije. Konverzija
3-hidroksi-2-metilpropanala iznosila je 77,33%, a iskorištenje na produktu 66,50%. Odabrani enzim kinetički je karakteriziran spektrofotometrijskom metodom, te su na temelju eksperimentalnih podataka procijenjene vrijednosti kinetičkih konstanti.

Abstract (english)

The oxidation reaction of 3-hydroxy-2-methylpropanal to the corresponding 3-hydroxy-2-methylpropanoic acid was carried out in this work. The main objective of the study was to search for an appropriate aldehyde dehydrogenase that can successfully oxidize the aldol product, from a given panel of 25 aldehyde dehydrogenases. As the product of aldol addition (also substrate - 3-hydroxy-2-methylpropanal) is not commercially available, it had to be synthesized prior to carrying out the oxidation reaction using the D-fructose-6-phosphate aldolase enzyme (FSA D6Q). Oxidation of aldol to the corresponding carboxylic acid is carried out using two enzymes (aldehyde dehydrogenase and NADH oxidase) and coenzyme NAD+. NADH oxidase (NOX) is an enzyme that translates NADH into NAD+, and is used to regenerate NAD+ coenzyme to minimize its consumption in the reactor. Aldehyde dehydrogenase 41 has proven to be the most successful biocatalyst for the oxidation reaction in the study. Conversion of 3-hydroxy-2-methylpropanal was 77,33%, and product yield 66,50%. The selected enzyme was kinetically characterized by a spectrophotometric method, and the values of kinetic constants were estimated based on experimental data.

• Keywords: aldehid-dehidrogenaza; aldolna adicija; oksidacija; enzim; koenzim; biokatalizator
• Keywords (english): aldehyde dehydrogenase; aldol addition; oxidation; enzyme; coenzyme; biocatalyst
• Language: croatian
• URN:NBN: urn:nbn:hr:149:162086
• 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: Applied organic chemistry; 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: Closed access
• Created on: 2023-05-26 10:34:12