Validacija modela biokatalitičke sinteze (R)-2-azido-1-(4-trifluorometil-fenil)-etanola u šaržnom reaktoru

master's thesis

• Title (english): Model validation for the biocatalytic synthesis of (R)-2-azido-1-(4-trifluoromethyl-phenyl)-ethanol in a batch reactor
• Author: Emanuel Tomljenović
• Mentor: Zvjezdana Findrik Blažević (mentor)
• Committee member: Zvjezdana Findrik Blažević (predsjednik povjerenstva)
• Committee member: Marija Vuković Domanovac (član povjerenstva)
• Committee member: Dragana Vuk (član povjerenstva)
• Granter: University of Zagreb; Faculty of Chemical Engineering and Technology; Zagreb
• Defense date and country: 2021-09-20, Croatia
• Scientific / art field, discipline and subdiscipline: TECHNICAL SCIENCES; Chemical Engineering

Abstract

Enzimi su biološki katalizatori koji su prisutni u živim stanicama i neophodni su za život kakav poznajemo. Reakcije u kojima su prisutni enzimi potrebno je dobro istražiti i okarakterizirati kako bi se mogle provoditi u industrijskim mjerilima. Prednost uporabe enzima u biokatalitičkim reakcijima leži u činjenici da su djelotvnorni i pri vrlo blagim uvjetima, puno blažim od kemijskih katalizatora. Potrebno je obratiti pažnju i na aktivnost enzima u različitim reakcijskim uvjetima (temperatura, koncentracija supstrata, pH, zasićenost kisikom) jer optimizacijom reakcijskih uvjeta omogućujemo maksimalan prinos željenog produkta i smanjenje nastanka neželjenog ili štetnog nusprodukta. Fluorirani aromatski spojevi su iznimno važni u farmaceutskoj industriji kao prekursori i kao građevni blokovi. Bitno je da takvi spojevi budu optički čisti jer je poznato da dva enantiomera mogu imati znatno različita svojstva i biološko djelovanje, često štetno i toksično. Jedna od metoda sinteze takvih spojeva uključuje korištenje enzima halogenhidrin-dehalogenaze (HHDH). U ovom radu proučavana je reakcija otvaranja epoksidnog prstena, fluoriranog derivata stiren oksida uz azid kao nukleofil prilikom čega nastaje fluorirani azidoalkohol. Reakcija je katalizirana enzimom HHDH (C-tip) koji je poznat i pod nazivom HheC-W249P. Cilj ovog rada bio je validirati matematički model biokatalitičke sinteze (R)-2-azido-1-(4-trifluorometil-fenil)-etanola u šaržnom reaktoru. Model je uspješno validiran provedbom reakcije u pet različitih kotlastih reaktora s različitim početnim uvjetima u reaktoru. Uzorci reakcijske smjese su analizirani na tekućinskom kromatografu visoke djelotvornosti. Praćena je i stabilnost enzima pri različitim uvjetima prilikom čega je pokazano da početni uvjeti u reaktoru ne utječu značajno na stabilnost enzima. Stabilnost enzima se pratila spektrofotometrijskom metodom.

Abstract (english)

Enzymes are biological catalysts that are present in living cells and are necessary for life as we know it. Reactions in which enzymes are present need to be well investigated and characterized in order to be carried out on an industrial scale. The advantage of using enzymes in biocatalytic reactions lies in the fact that they are effective even under very mild conditions, much milder than catalysts of chemical reactions. It is necessary to pay attention to the activity of enzymes in different reaction conditions (temperature, substrate concentration, pH, oxygen saturation) because by optimizing the reaction conditions we enable maximum yield of the desired product and reduction of unwanted or harmful by-products. Fluorinated aromatic compounds are extremely important in the pharmaceutical industry as precursors and as building blocks. It is important that such compounds be optically pure because it is known that the two enantiomers can have significantly different properties and biological effect, often harmful and toxic. One of the methods of synthesis of such compounds is the use of the enzyme halohydrin-dehalogenase (HHDH). In this paper, the reaction of opening an epoxy ring, a fluorinated derivative of styrene oxide with an azide as a nucleophile, during which a fluorinated azidoalcohol is formed, was studied. The reaction is catalyzed by the HHDH (C-type) enzyme also known as HheC-W249P. The aim of this work was to validate a mathematical model of biocatalytic synthesis of
(R)-2-azido-1-(4-trifluoromethyl-phenyl)-ethanol in a batch reactor. The model was successfully validated by carrying out the reaction in five different batch reactors with different initial conditions in the reactor. Samples of the reaction mixture were analyzed on a high performance liquid chromatograph. The stability of the enzyme under different conditions was also monitored, and it was shown that the initial conditions in the reactor did not significantly affect the stability of the enzyme. Enzyme stability was monitored by spectrophotometric method.

• Keywords: enzimi; biokatalitičke reakcije; halogenhidrin-dehalogenaza (HHDH); epoksidi
• Keywords (english): enzymes; biocatalytic reactions; halohydrin-dehalogenase (HHDH); epoxides
• Language: croatian
• URN:NBN: urn:nbn:hr:149:954362
• Project: Number: IP-2018-01-4493; Title: Enzimska sinteza fluoriranih kiralnih građevnih blokova; Acronym: EnzyFluor; Leader: Maja Majerić-Elenkov; Jurisdiction: Croatia; Funder: HRZZ; Funding stream: IP
• Study programme: Title: Chemical Engineering - Graduate study; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka kemijskog inženjerstva (magistar/magistra inženjer/inženjerka kemijskog inženjerstva)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access; Embargo expiration date: 2022-09-01
• Created on: 2023-04-14 10:51:08