Aktivnost ketoreduktaza pri oksidaciji diola

undergraduate thesis

• Title (english): Activity of ketoreductases during diol oxidation
• Author: Valerija Matković
• Mentor: Zvjezdana Findrik Blažević (mentor)
• Committee member: Zvjezdana Findrik Blažević (predsjednik povjerenstva)
• Committee member: Martina Sudar (član povjerenstva)
• Committee member: Marija Vuković Domanovac (član povjerenstva)
• Granter: University of Zagreb; Faculty of Chemical Engineering and Technology; Zagreb
• Defense date and country: 2017-09-21, Croatia
• Scientific / art field, discipline and subdiscipline: TECHNICAL SCIENCES; Chemical Engineering; Reaction Engineering

Abstract

U radu je opisana kaskadna reakcija koja objedinjuje reakciju oksidacije Cbz-N-3-amino-1,2-propandiola (diol) te reakciju aldolne adicije dihidroksiacetona (DHA) na aldehid sintetiziran u koraku oksidacije. Za reakciju oksidacije testirano je 18 ketoreduktaza, te je njihova aktivnost uspoređena s aktivnošću enzima alkohol dehidrogenaze iz jetre konja (HLADH) koji se ranije pokazao kao katalizator aktivan u proučavanoj oksidoredukciji. Ketoreduktaze i dehidrogenaze zahtijevaju prisutnost koenzima NAD+ ili NADP+. Među testiranim enzimima pronađena su 2 aktivna uz NAD+, te tri aktivna uz koenzim NADP+ u proučavanoj oksidoredukciji. Odabrani enzimi su korišteni za provedbu kaskadne reakcije u kojoj je nakon oksidoredukcije slijedila aldolna adicija dihidroksiacetona na aldehid katalizirana D-fruktoza-6-fosfat aldolazom (FSA). Za učinkovitu oksidaciju diola korištena je NADH oksidaza (NOX) za regeneraciju koenzima. Kaskadna reakcija je praćena preko koncentracije nastalog aldolnog produkta nakon 24 i 48 sati, koja je ujedno bila korištena i kao mjerilo uspješnosti provedene reakcije. Usprkos tomu što je u provedenim reakcijama nastao željeni produkt, zaključeno je da je među ispitanima HLADH najbolji katalizator za navedenu reakciju.

Abstract (english)

This paper describes a cascade reaction which combines the oxidation reaction of Cbz-N-3-amino-1,2-propanediol (diol) and the reaction of the aldol addition of dihydroxyacetone (DHA) to the aldehyde synthesized in the oxidation step. 18 ketoreductases were tested for oxidation reaction, and their activity was compared with the activity of alcohol dehydrogenase from horse liver (HLADH), which previously proved to be the catalyst active in the study of oxidation. Ketoreductases and dehydrogenases require the presence of NAD+ or NADP+ coenzyme. Among the tested enzymes, 2 were active in the studied reaction with NAD+, and three with coenzyme NADP+. The selected enzymes were used to carry out a cascade reaction in which following oxidation, the aldol addition of dihydroxyacetone to aldehyde catalyzed by D-fructose-6-phosphate aldolase (FSA) was carried out. NADH oxidase (NOX) for coenzyme regeneration was used for the effective oxidation of diol. The concentration of the formed aldol product after 24 and 48 hours in the cascade reaction was followed and was used as a measure of the reaction effectiveness. In spite of the fact that the desired product was formed in all reactions, HLADH was still found to be the best catalyst for this reaction.

• Keywords: kaskadna reakcija; aldolna adicija; Cbz-N-3-amino-1 2-propandiol; ketoreduktaze; NADH oksidaza
• Keywords (english): cascade reaction; aldol addition; Cbz-N-3-amino-1 2-propandiol; ketoreductases; NADH oxidase
• Language: croatian
• URN:NBN: urn:nbn:hr:149:514634
• 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: Chemical Engineering - Undergraduate study; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/ prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka kemijskog inženjerstva (sveučilišni/a prvostupnik/ prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka kemijskog inženjerstva)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2021-03-23 09:53:17