Trbojević Ivić Jovana, Veličković Dušan, Dimitrijević Aleksandra, Bezbradica Dejan, Dragačević Vladimir, Gavrović Jankulović Marija, Milosavić Nenad
Innovation Center, Faculty of Chemistry, University of Belgrade, 11000, Belgrade, Serbia.
Department of Biochemistry, Faculty of Chemistry, University of Belgrade, 11000, Belgrade, Serbia.
J Sci Food Agric. 2016 Sep;96(12):4281-7. doi: 10.1002/jsfa.7641. Epub 2016 Feb 26.
Biocatalysts are a promising alternative for the production of natural flavor compounds. Candida rugosa lipase (CRL) is a particularly important biocatalyst owing to its remarkable efficiency in both hydrolysis and synthesis. However, additional stabilization is necessary for successful industrial implementation. This study presents an easy and time-saving method for immobilizing this valuable enzyme on hydroxyapatite (HAP), a biomaterial with high protein-binding capacity.
Targeted immobilized CRL was obtained in high yield of ≥98%. Significant lipase stabilization was observed upon immobilization: at 60 °C, immobilized lipase (HAP-CRL) retained almost unchanged activity after 3 h, while free CRL lost 50% of its initial activity after only 30 min. The same trend was observed with tested organic solvents. Methanol and hexane had the most pronounced effect: after 3 h, only HAP-CRL was stable and active, while CRL was completely inactivated. The practical value of the prepared catalyst was tested in the synthesis of the aroma ester methyl acetate in hexane. Reaction yields were 2.6 and 52.5% for CRL and HAP-CRL respectively.
This research has successfully combined an industrially prominent biocatalyst, CRL, and a biocompatible, environmentally suitable carrier, HAP, into an immobilized preparation with improved catalytic properties. The obtained CRL preparation has excellent potential for the food and flavor industries, major consumers in the global enzyme market. © 2016 Society of Chemical Industry.
生物催化剂是生产天然香料化合物的一种有前景的替代方法。皱落假丝酵母脂肪酶(CRL)是一种特别重要的生物催化剂,因为它在水解和合成方面都具有显著的效率。然而,为了成功实现工业化应用,还需要进一步提高其稳定性。本研究提出了一种简便省时的方法,将这种有价值的酶固定在具有高蛋白结合能力的生物材料羟基磷灰石(HAP)上。
以≥98%的高产率获得了靶向固定化的CRL。固定化后观察到脂肪酶有显著的稳定性:在60°C下,固定化脂肪酶(HAP-CRL)在3小时后活性几乎保持不变,而游离CRL仅在30分钟后就失去了50%的初始活性。在测试的有机溶剂中也观察到了相同的趋势。甲醇和己烷的影响最为显著:3小时后,只有HAP-CRL保持稳定且有活性,而CRL则完全失活。在己烷中合成香气酯乙酸甲酯时测试了制备的催化剂的实际价值。CRL和HAP-CRL的反应产率分别为2.6%和52.5%。
本研究成功地将一种工业上突出的生物催化剂CRL和一种生物相容、环境适宜的载体HAP结合成一种具有改进催化性能的固定化制剂。所获得的CRL制剂在食品和香料工业(全球酶市场的主要消费领域)具有极好的应用潜力。©2016化学工业协会。