The effect of deep eutectic solvents on lipase catalyzed hydrolysis of (R,S)-1-phenylethylcetate

Posljednjih nekoliko godina raste broj istraživanja usmjerenih na mogućnost primjene eutektičkih otapala u različitim biotransformacijskim procesima. Glavne prednosti njihove primjene u odnosu na ostala otapala su biorazgradivost, netoksičnost, nehlapljivost te jednostavna priprava. U ovom radu ispitana je mogućnost primjene imobilizirane lipaze te hidrolitičkih enzima iz narančine kore kao biokatalizatora u hidrolizi (R,S)-1-feniletil-acetata u prirodnom eutektičkom otapalu kolin-klorid:etilen-glikol (ChClEG) s različitim udjelima vode. Uspoređujući postotak iskorištenja hidrolize i enantiomernog viška, utvrđeno je da je najpogodnije otapalo ChClEG s 50 % udjela vode (w/w). Također, ispitana je skladišna stabilnost imobilizirane lipaze B te hidrolitičkih enzima narančine kore kroz vremenski period od 21 dan u eutektičkom otapalu u ChClEG. Iz dobivenih vrijednosti rezidualne aktivnosti utvrđeno je da su hidrolitički enzimi iz narančine kore stabilniji u ChClEG nego u puferu, dok je lipaza stabilnija u puferu nego u ChClEG s 50 % udjela vode (w/w).In the last few years, there has been an increasing number of studies focused on the possibilities of using eutectic solvents in various biotransformation processes. The main advantages of their application over other solvents are biodegradability, non-toxicity, non-volatility and easy synthesis. This work examines the possibility of using immobilized lipase and hydrolytic enzymes from orange peel as biocatalysts in the hydrolysis of (R,S)-1-phenylethyl acetate in a natural deep eutectic solvent choline chloride:ethylene glycol with different water contents. Comparing the percentage of utilization of hydrolysis and enantiomeric excess, it was found that the most suitable solvent is ChClEG with 50% water content (w/w). Also, the storage thermal stability of immobilized lipase and hydrolytic enzymes from orange peel over a period of 21 days was examined. From the obtained residual activity values, it was found that the hydrolytic enzymes from the orange peel were more stable in ChClEG than in buffer, while lipase was more stable in buffer than in ChClEG with 50% water content (w/w)