Enzimski katalizirana kaskadna reakcija za sintezu prekursora D-fagomina

master's thesis

• Title (english): Enzymatic cascade reaction for the synthesis of D-fagomine precursor
• Author: Mariana Klepić
• 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: 2021-09-24, Croatia
• Scientific / art field, discipline and subdiscipline: TECHNICAL SCIENCES; Chemical Engineering

Abstract

U ovom radu proučavana je enzimski katalizirana kaskadna reakcija za sintezu
(3S,4R)-6-[(benziloksicarbonil)amino]-5,6-dideoksiheks-2-uloze, prekursora D-fagomina. U reakciji sudjeluju tri enzima, alkohol dehidrogenaza iz konjske jetre koja oksidira
N-Cbz-3-aminopropanol u odgovarajući aldehid, NADH oksidaza iz Lactococcus lactis koja se koristi za regeneraciju koenzima NAD+ i D-fruktoza 6-fosfat aldolaza iz Escherichia coli za aldolnu adiciju dihidroksiacetona na N-Cbz-3-aminopropanal. Mjerena je specifična aktivnost enzima NADH oksidaze tijekom vremena s i bez flavin adenin dinukleotida te je procijenjena konstanta deaktivacije enzima. Procijenjen je i koeficijent prijenosa kisika u reakcijskoj smjesi s i bez aeracije. Razvijen je matematički model ispitivane reakcije te su provedene simulacije u kojima su ispitani utjecaji koncentracija enzima NADH oksidaze i alkohol dehidrogenaze, 𝑘𝐿𝑎 i 𝑘𝑑 na konverziju N-Cbz-3-aminopropanola, iskorištenja na aldolu i kiselini, te utjecaj
𝑘𝑑 NOX-a na brzinu regeneracije koenzima i koncentraciju NAD+ tijekom provedbe reakcije.

Abstract (english)

Enzymatic cascade reaction is studied for the synthesis of
(3S,4R)-6-[(benzyloxycarbonyl)amino]-5,6-dideoxyhex-2-ulose, D-fagomine precursor. Reaction involves three enzymes, horse liver alcohol dehydrogenase for the oxidation of N-Cbz-3-aminopropanol to the corresponding aldehyde, NADH oxidase from Lactococcus lactis is used for the regeneration of coenzyme NAD+ and D-fructose-6-phosphate aldolase from Escherichia coli for the aldol addition of dihydroxyacetone to N-Cbz-3-aminopropanal. Specific activity of the enzyme NADH oxidase was measured with and without flavin adenine dinucleotide and the operational stability decay rate constant of the enzyme was estimated. Oxygen transfer coefficient in the reaction mixture was also determined with and without aeration. Mathematical model of the studied reaction was developed and different simulations were carried out. Simulations show the influence of NADH oxidase concentrations, alcohol dehydrogenase, 𝑘𝐿𝑎 and 𝑘𝑑 on the conversion of
N-Cbz-3-aminopropanol, yield of the aldol and acid, as well as the influence of 𝑘𝑑 of NOX on the reaction rate of coenzyme regeneration and NAD+ concentration during the reaction.

• Keywords: enzimska kataliza; kaskadna reakcija; aeracija
• Keywords (english): enzyme catalysis; cascade reaction; aeration
• Language: croatian
• URN:NBN: urn:nbn:hr:149:432152
• Study programme: Title: Environmental Engineering - Graduate study; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka ekoinženjerstva (magistar/magistra inženjer/inženjerka ekoinženjerstva)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Closed access
• Created on: 2023-04-17 11:54:11