Proizvodnja optički aktivnog diola korištenjem višeenzimskog sustava

master's thesis

• Title (english): Production of optically active diols by multienzyme catalysis approach
• Author: Lela Pintarić
• Mentor: Ana Vrsalović Presečki (mentor)
• Committee member: Ana Vrsalović Presečki (predsjednik povjerenstva)
• Committee member: Zvjezdana Findrik Blažević (član povjerenstva)
• Committee member: Ana Lončarić Božić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Chemical Engineering and Technology; Zagreb
• Defense date and country: 2015-07-13, Croatia
• Scientific / art field, discipline and subdiscipline: TECHNICAL SCIENCES; Chemical Engineering; Reaction Engineering

Abstract

Enantiočisti kiralni alkoholi koriste se za sintezu različitih industrijski važnih kemikalija, kao što su lijekovi, herbicidi, insekticidi i sl. Jedan od najvažnijih načina njihove proizvodnje je asimetrična redukcija ketona. U ovom je radu, reakcijom benzaldehida i acetaldehida kataliziranom enzimom benzoilformat dekarboksilazom (BFD) iz bakterije Pseudomonas putida, sintetiziran (S)-2-hidroksipropiofenon ((S)-2-HPP). Iz njega je zatim proizveden (1S,2S)-1-fenilpropan-1,2-diol ((1S,2S)-1-FPD), a reakcija je katalizirana enzimom alkohol dehidrogenazom iz bakterije Lactobacillus brevis (LbADH). Reakcija je provedena uz regeneraciju koenzima nikotinamid adenin dinukleotida, NADH, koja je ostvarena dodatkom amonij formijata i enzima formijat dehidrogenaze iz kvasca Candida boidinii (CbFDH).
Kinetika enzima BFD opisana je dvosupstratnom Michaelis – Menteničinom kinetikom s kompetitivnom inhibicijom supstratom, acetaldehidom. Kinetika enzima LbADH uz prisustvo koenzima NADH opisana je dvosupstratnom Michaelis – Menteničinom kinetikom s kompetitivnom inhibicijom produktom, NAD+, dok je kinetika povratne reakcije opisana kinetikom drugog reda. Kinetika enzima CbFDH opisana je dvosupstratnom Michaelis – Menteničinom kinetikom s kompetitivnom inhibicijom produktom, NADH, s uključenom inhibicijom supstratima iz drugih reakcija, (S)-2-HPP i acetaldehidom.
Postavljen je matematički model procesa koji je validiran u kotlastom reaktoru. Usporedno su provedena dva eksperimenta s različitim početnim koncentracijama acetaldehida. Zaključeno je kako je eksperiment bolje provoditi uz više koncentracije acetaldehida, s obzirom na kraće trajanje reakcije i činjenicu da je deaktivacija korištenih enzima u oba slučaja podjednaka i neovisna o koncentraciji acetaldehida.

Abstract (english)

Enantiopure chiral alcohols are used for synthesis of various industrially important chemicals, such as medicines, herbicides, insecticides, etc. One of the most important methods used to produce them is asymmetric reduction of ketones. In this study, (S)-2-hydroxypropiophenone ((S)-2-HPP) was synthesized from benzaldehyde and acetaldehyde. The reaction was catalyzed by the enzyme benzoylformate decarboxylase (BFD) from bacteria Pseudomonas putida. Then, (1S,2S)-1-phenylpropane-1,2-diol ((1S,2S)-1-FPD) was synthesized from (S)-2-HPP. This reaction was catalyzed by the enzyme alcohol dehydrogenase from bacteria Lactobacillus brevis (LbADH). Simultaneously, regeneration of nicotinamide adenine dinucleotide coenzyme (NADH) was carried out by addition of ammonium formate and enzyme formate dehydrogenase from yeast Candida boidinii (CbFDH).
The BFD kinetics was described by the double substrate Michaelis – Menten model with a competitive inhibition by the substrate, acetaldehyde. The LbADH kinetics was described by double substrate Michaelis – Menten model with a competitive inhibition by product, NAD+, while the second-order model describes the reverse reaction kinetics. The CbFDH kinetics was described by double substrate Michaelis – Menten model with a competitive inhibition by product, NADH, and substrates from main reactions, (S)-2-HPP and acetaldehyde.
The mathematical model was developed and validated in a batch reactor. Two experiments with different initial concentrations of acetaldehyde were carried out. It is concluded that it is better to carry out experiments with higher acetaldehyde concentrations, considering shorter reaction time and the fact that the deactivation of the enzymes is equal in both cases and independent on the acetaldehyde concentration.

• Keywords: benzoilformat dekarboksilaza; alkohol dehidrogenaza; formijat dehidrogenaza; (1S; 2S)-1-fenilpropan-1 2-diol
• Keywords (english): benzoylformate decarboxylase; alcohol dehydrogenase; formate dehydrogenase; (1S; 2S)-1-phenylpropane-1 2-diol
• Language: croatian
• URN:NBN: urn:nbn:hr:149:430829
• 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: 2016-12-16 13:46:47