Optimiranje kaskadne reakcije sinteze (3S,4R)-5-[N-(benziloksikarbonil)amino]-5-deoksipent-2-uloze

master's thesis

• Title (english): Optimization of cascade synthesis of (3S,4R)-5-[N-(benzyloxycarbonyl)amino]-5-deoxypent-2-ulose
• Author: Anamarija Slivar
• Mentor: Đurđa Vasić-Rački (mentor)
• Committee member: Đurđa Vasić-Rački (predsjednik povjerenstva)
• Committee member: Zvjezdana Findrik Blažević (član povjerenstva)
• Committee member: Irena Škorić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Chemical Engineering and Technology; Zagreb
• Defense date and country: 2015-09-22, Croatia
• Scientific / art field, discipline and subdiscipline: TECHNICAL SCIENCES; Chemical Engineering; Reaction Engineering

Abstract

U ovom radu je proučavana sinteza (3S, 4R)-5-[N-(benziloksikarbonil)amino]deoksipent-2-uloze u kaskadnoj reakciji oksidacije etanolamina kataliziranoj alkohol dehidrogenazom koja je izolirana iz konjske jetre uz regeneraciju koenzima kataliziranom NADH oksidazom i aldolne adicije dihidroksiacetona (DHA) na N-Cbz-glicinal kataliziranoj D-fruktoza-6-fosfat aldolazom (FSA A129N) porijeklom iz Escherichia coli. Kinetika reakcije aldolne adicije DHA na N-Cbz-glicinal i oksidacije etanolamina ispitana je metodom početnih brzina reakcije. Ispitan je utjecaj koncentracija supstrata (DHA, N-Cbz-glicinala) i produkta (aldola) na početnu brzinu reakcije aldolne adicije. Kako se radi o povratnoj reakciji ispitan je i utjecaj koncentracija supstrata (aldola) i produkta (DHA, N-Cbz-glicinala) na početnu brzinu retro-aldolne reakcije. Kod reakcije oksidacije etanolamina ispitan je utjecaj supstrata (etanolamina i NAD+) na početnu brzinu reakcije oksidacije. Iz eksperimentalnih podataka procijenjeni su kinetički parametri metodom nelinearne regresije, te su razvijeni matematički modeli reakcija u kotlastom reaktoru. Razvijen matematički model u kotlastom reaktoru za reakciju aldolne adicije DHA na N-Cbz-glicinal validiran je eksperimentalno i dobro opisuje eksperimentalne podatke. Razvijen je i matematički model kaskadne reakcije oksidacije etanolamina katalizirane s ADH uz regeneraciju koenzima kataliziranu NADH oksidazom i aldolne adicije DHA na N-Cbz-glicinal katalizirane s FSA A129N, uz pomoć kojeg je provedena optimizacija kaskadne reakcije. Varirane su koncentracije enzima FSA, ADH, NOX-a i koenzima NAD+ kako bi se dobilo najveće iskorištenje na konačnom produktu. Optimizacijom kaskadne reakcije uz pomoć razvijenog matematičkog modela dobiveno je najveće iskorištenje od 77,96 % pri koncentraciji FSA od 0,5 mg/cm^3, HLADH od 15 mg/cm^3, NOX od 0,1 mg/cm^3 i NAD+ od 1,0 mmol/dm^3.

Abstract (english)

The synthesis of (3S, 4R)-5-[N-(benzyloxycarbonyl)amino]deoxypent-2-ulose in a cascade reaction of ethanolamine oxidation catalyzed by alcohol dehydrogenase isolated from horse liver with coenzyme regeneration catalyzed with NADH oxidase and aldol addition of dihydroxyacetone (DHA) to N-Cbz-glycinal catalyzed by D-fructose-6-phosphate aldolase (FSA A129N) from Escherichia coli was studied in this paper. Reaction kinetics of aldol addition of DHA to N-Cbz-glycinal and ethanolamine oxidation were investigated using initial reaction rate method. The influence of substrate (DHA, N-Cbz-glycinal) and product concentration (aldol) on the initial reaction rate of aldol addition was investigated. Since this is an equilibirum reaction, the influence of substrate (aldol) and product concentration (DHA, N-Cbz-glycinal) on the inital reaction rate of the retro-aldol reaction was also investigated. The influence of substrate concentration (ethanolamine, NAD+) on the initial reaction rate in the reaction of ethanolamine oxidationwas investigated. Kinetic parameters were estimated from the experimental data by the non-linear regression method and mathematical models of reactions in batch reactor were developed. Developed mathematical model in the batch reactor for the aldol addition of DHA to N-Cbz-glycinal was validated experimentally and it described experimental data well. Mathematical model for the cascade reaction of ethanolamine oxidation catalyzed with ADH with coenzyme regeneration catalyzed with NADH oxidase and aldol addition of DHA to N-Cbz-glycinal catalyzed with FSA A129N was developed. Furthermore, optimization of this cascade reaction was conducted. Concentrations of enzymes FSA, ADH, NOX and coenzyme NAD+ are varied to give the maximum yield of the product. Optimization of the cascade reaction with the help of the developed mathematical model gave the highest yield of 77.96 % at concentrations of FSA at 0.5 mg/cm^3, HLADH 15 mg/cm^3, NOX 0.1 mg/cm^3 and NAD+ 1.0 mmol/dm^3.

• Keywords: kaskadna reakcija; aldolna adicija; oksidacija; matematički model; optimizacija
• Keywords (english): cascade reaction; aldol addition; oxidation; mathematical model; optimization
• Language: croatian
• URN:NBN: urn:nbn:hr:149:142290
• 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: Open access
• Created on: 2017-03-06 14:20:13