Ben Rejeb Ines, Arduini Fabiana, Amine Aziz, Gargouri Mohamed, Palleschi Giuseppe
Biocatalysis and Bioprocess Unit, National Institute of Applied Science and Technology (INSAT), B.P. 676, 1080 Tunis Cedex, Tunisia.
Anal Chim Acta. 2007 Jun 26;594(1):1-8. doi: 10.1016/j.aca.2007.04.066. Epub 2007 May 10.
Lipase activity against triacylglycerols has been measured using an amperometric enzyme biosensor based on glycerol dehydrogenase/NADH oxidase. A Prussian Blue modified screen-printed electrode was selected as substrate for the two immobilised-enzyme systems due to their higher operative stability reported in previous works. Various parameters such as cofactor (flavin mononucleotide FMN) concentration (1 mM), NAD+ coenzyme concentration (2 mM), pH effect (phosphate buffer pH 6 to 8, Tris buffer pH 8-10) response time and storage stability were evaluated and optimised. The glycerol biosensor was then investigated for lipase activity. The system was challenged against an olive or sunflower oil real samples in order to detect fatty acids and the results were compared with those provided either by the manufacture or reference methods with good agreement.
已使用基于甘油脱氢酶/ NADH氧化酶的安培型酶生物传感器测量了针对三酰甘油的脂肪酶活性。由于先前工作中报道的更高的操作稳定性,选择普鲁士蓝修饰的丝网印刷电极作为两种固定化酶系统的底物。评估并优化了各种参数,如辅因子(黄素单核苷酸FMN)浓度(1 mM)、NAD +辅酶浓度(2 mM)、pH效应(磷酸盐缓冲液pH 6至8,Tris缓冲液pH 8 - 10)、响应时间和储存稳定性。然后研究了甘油生物传感器的脂肪酶活性。该系统针对橄榄油或葵花籽油实际样品进行测试以检测脂肪酸,结果与制造商提供的结果或参考方法进行比较,吻合度良好。