Milašinović Nikola, Jakovetić Sonja, Knežević-Jugović Zorica, Milosavljević Nedeljko, Lučić Marija, Filipović Jovanka, Kalagasidis Krušić Melina
Department of Criminalistic Sciences, The Academy of Criminalistic and Police Studies, Cara Dušana 196, 11080 Belgrade, Serbia.
Department of Biochemical Engineering and Biotechnology, Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, 11000 Belgrade, Serbia.
ScientificWorldJournal. 2014 Feb 20;2014:142123. doi: 10.1155/2014/142123. eCollection 2014.
This study reports the synthesis of polymeric matrices based on N-isopropylacrylamide and itaconic acid and its application for immobilization of lipase from Candida rugosa. The lipase was immobilized by entrapment method. Free and immobilized lipase activities, pH and temperature optima, and storage stability were investigated. The optimum temperature for free and entrapped lipase was found to be 40 and 45 °C, while the optimum pH was observed at pH 7 and 8, respectively. Both hydrolytic activity in an aqueous medium and esterolytic activity in an organic medium have been evaluated. Maximum reaction rate (V max) and Michaelis-Menten constants (K m ) were also determined for immobilized lipase. Storage stability of lipase was increased as a result of immobilization process. Furthermore, the operational stability and reusability of the immobilized lipase in esterification reaction have been studied, and it was observed that after 10 cycles, the residual activity for entrapped lipase was as high as 50%, implying that the developed hydrogel and immobilized system could provide a promising solution for the flavor ester synthesis at the industrial scale.
本研究报道了基于N-异丙基丙烯酰胺和衣康酸的聚合物基质的合成及其用于固定化皱褶假丝酵母脂肪酶的应用。脂肪酶通过包埋法固定化。研究了游离和固定化脂肪酶的活性、最适pH和温度以及储存稳定性。发现游离和包埋脂肪酶的最适温度分别为40和45℃，而最适pH分别在pH 7和8时观察到。评估了在水介质中的水解活性和在有机介质中的酯解活性。还测定了固定化脂肪酶的最大反应速率（V max）和米氏常数（K m）。固定化过程提高了脂肪酶的储存稳定性。此外，研究了固定化脂肪酶在酯化反应中的操作稳定性和可重复使用性，观察到在10个循环后，包埋脂肪酶的残余活性高达50%，这意味着所开发的水凝胶和固定化系统可为工业规模的风味酯合成提供有前景的解决方案。
