Utjecaj eutektičkih otapala na aktivnost alkohol dehidrogenaze i stabilnost nikotinamidnih koenzima

undergraduate thesis

• Title (english): Effects of deep eutectic solvents on alcohol dehydrogenase activity and stability of nicotinamide coenzymes
• Author: Romana Ivković
• Mentor: Marina Cvjetko Bubalo (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for Cell Technology, Application and Biotransformations); Zagreb
• Defense date and country: 2020-09-15, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Gotovo svaki industrijski proces proizvodnje uključuje primjenu organskih otapala čija je uporaba odgovorna za onečišćenje okoliša te nepovoljna za zdravlje radnika i sigurnost procesa zbog svojstva toksičnosti, lake hlapljivosti, zapaljivosti i eksplozivnosti. Stoga, kako bi se ispunili zahtjevi zelene kemije za ekološkom prihvatljivošću i održivošću, traže se alternativna otapala. Eutektička otapala istaknula su se svojstvima poput niske hlapljivosti, netoksičnosti, nezapaljivosti, biorazgradivosti te jednostavnim metodama priprave. Nadalje, u polju biokatalize dehidrogenaze pokazuju veliki potencijal za dobivanje visoko vrijednih kiralnih spojeva koji se koriste u proizvodnji optički čistih lijekova. Cilj ovog rada bio je istražiti mogućnost primjene prirodnih eutektičkih otapala na bazi kolin-klorida kao medija za reakcije redukcije katalizirane dehidrogenazama. U pripravljenim otapalima određena je aktivnost alkohol dehidrogenaze te promjene u konformaciji iste. U istim otapalima također je ispitana stabilnost pripadajućih koenzima NADH/NAD+. Blago lužnata prirodna eutektička otapala koja su sadržavala etilen-glikol i ureu pokazala su stabilizirajući utjecaj na oba koenzima, no niti u jednom otapalu alkohol dehidrogenaza nije pokazala zadovoljavajuću aktivnost uz značajne promjene u konformaciji u odnosu na pufer kao referentno otapalo.

Abstract (english)

Almost every industrial production process involves the use of organic solvents whose use is responsible for environmental pollution and unfavorable to workers' health and process safety, due to the properties of toxicity, volatility, flammability and explosiveness. Therefore, in order to meet the requirements of green chemistry for environmental safety and sustainability, alternative solvents are sought. Eutectic solvents stood out with properties such as low volatility, non-toxicity, non-flammability, biodegradability and simple preparation methods. Furthermore, in the field of biocatalysis, dehydrogenases show great potential for obtaining high-value chiral compounds used in the production of optically pure drugs. The aim of this study was to investigate the possibility of using natural eutectic solvents based on choline chloride as a medium for dehydrogenase-catalyzed reduction reactions. The activity of alcohol dehydrogenase and changes in its conformation was determined in the prepared solvents. The stability of the corresponding NADH/NAD+ coenzymes was also tested in the same solvents. Slightly alkaline natural eutectic solvents containing ethylene glycol and urea showed a stabilizing effect on both coenzymes, but in neither solvent did alcohol dehydrogenase show satisfactory activity with significant changes in conformation relative to buffer as reference solvent.

• Keywords: biokatalizatori; dehidrogenaze; eutektička otapala; nikotinamidni koenzimi; zelena kemija
• Keywords (english): biocatalysts; deep eutectic solvents; dehydrogenases; green chemistry; nicotinamide coenzyme
• Language: croatian
• URN:NBN: urn:nbn:hr:159:561674
• Study programme: Title: Biotechnology; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2020-10-29 12:08:54