Stabilnost nitrilaze pri sintezi (R)-4-klor-3-hidroksibutanske kiseline

undergraduate thesis

• Title (english): Stability of nitrilase during synthesis of (R)-4-chloro-3-hydroxybutyric acid
• Author: Martina Miloščić
• 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: 2023-09-04, Croatia
• Scientific / art field, discipline and subdiscipline: TECHNICAL SCIENCES; Chemical Engineering

Abstract

Enzimi, odnosno biokatalizatori, imaju brojne prednosti nad kemijskim katalizatorima te se zbog toga njihova upotreba i potražnja u farmaceutskoj industriji neprestano povećava. Enzimi koji kataliziraju reakcije hidrolize nitrila u karboksilne kiseline i amonijak bez stvaranja amidnih međuprodukata nazivaju se nitrilazama. Nitrilaza se koristi kao biokatalizator u reakcijama pretvorbe (R)-4-klor-3-hidroksibutironitrila u (R)-4-klor-hidroksibutansku kiselinu. Zatim dolazi do spontane ciklizacije (R)-4-klor-3-hidroksibutanske kiseline u
(R)-3-hidroksi-γ-butirolakton koji je svestrani kiralni gradivni blok, a često se koristi u sintezama mnogih farmaceutskih polimera i otapala. U ovome se radu pratila stabilnost nitrilaze pri sintezi (R)-4-klor-hidroksibutanske kiseline. Sinteza (R)-4-klor-hidroksibutanske kiseline provedena je u kotlastom reaktoru. Spektrofotometrijskim mjerenjima određivana je aktivnost nitrilaze pri različitim koncentracijama supstrata, (R)-4-klor-3-hidroksibutironitrila, kroz vremensko razdoblje od 24 h te su uspoređeni eksperimentalno dobiveni podaci s onima predviđenim matematičkim modelom. Uočeno je da aktivnost nitrilaze pada s vremenom, odnosno dolazi do deaktivacije nitrilaze. Iz dobivenih eksperimentalnih podataka procijenjena je konstanta deaktivacije pri različitim koncentracijama supstrata i enzima te je zaključeno kako početni uvjeti u reaktoru utječu na stabilnost nitrilaze.

Abstract (english)

Enzymes, also known as biocatalysts, have numerous advantages over chemical catalysts and because of that their use and demand in pharmaceutical industry is constantly growing. The enzymes that catalyze reactions of nitrile hydrolisis into carboxylic acids and ammonia without the formation of amide intermediates are called nitrilases. Nitrilases are used in conversion reactions (R)-4-chlorine-3-hydroxybutyronitrile into
(R)-4-chlorine-3-hydroxybutyric acid. Afretwords, a spontaneous reaction of cyclisation (R)-4-chlorine-3-hydroxybyric acid into (R)-3-hydroxy-γ-butyrolactone occurs, a versatile chiral building block, often used in synthesis of numerous pharmaceutical polymers and solutions. In this paper, the stability of nitrilase during the synthesis of
(R)-4-chlorine-3-hydroxybutyric acid was monitored. The synthesis of
(R)-4-chlorine-3-hydroxybutyric acid takes place in a batch reactor. The activity of nitrilase at different contrentrations of the substrate, (R)-4-chlorine-3-hydroxybutyronitrile, over a period of 24 hours was measured with the spectrophotometric assay. The experimentally obtained data was compared with those predicted by the mathematical model. It was observed that the activity of nitrilase decreases with time and it was concluded that deactivation of nitrilase occurs. The deactivation constant at different substrate and enzyme concentration was estimated from the obtained experimental data and it was concluded that the initial conditions in the reactor affect the stability of nitrilase.

• Keywords: enzimi; stabilnost enzima; nitrilaza; (R)-4-klor-3-hidroksibutironitril; (R)-4-klor-hidroksibutanska kiselina; (R)-3-hidroksi-γ-butirolakton; kotlasti reaktor
• Keywords (english): enzymes; stability of enzymes; nitrilase; (R)-4-chlorine-3-hydroxybutyronitrile; (R)-4-chlorine-3-hydroxybutyric acid; (R)-3-hydroxy-γ-butyrolactone; batch reactor
• Language: croatian
• URN:NBN: urn:nbn:hr:149:258301
• Study programme: Title: Applied Chemistry - Undergraduate study; specializations in: Environmental chemistry and green technologies, Advanced materials and technologies, Applied organic chemistry; Course: Applied organic chemistry; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) primijenjene kemije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) primijenjene kemije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Embargoed access; Embargo expiration date: 2026-09-01
• Created on: 2023-09-06 08:31:27