Institute of Biochemistry and Biotechnology University of the Punjab, Lahore, Pakistan; Institute of Molecular Biology and Biotechnology University of the Lahore, Lahore 58810, Pakistan.
Institute of Biochemistry and Biotechnology, Quaid-i-Azam Campus, University of the Punjab, Lahore 54590, Pakistan.
Food Chem. 2022 Mar 15;372:131317. doi: 10.1016/j.foodchem.2021.131317. Epub 2021 Oct 6.
Pectinases hydrolyze pectin and make up 25% of global food processing enzyme sales. In this study, we aimed to purify exo-polygalacturonase (Exo-PG) by using galacturonic acid conjugated magnetic nanoparticles (MNPs) and examined its application in juice purification. The submerged fermentation was carried out in the presence of apple pectin (1%) to promote production of exo-PG from Aspergillus flavus. Maximum exo-PG activity was observed after 4 days (30 °C and pH 5.0). A single protein band (66 kDa) of purified exo-PG was observed in SDS-PAGE. Purification of exo-PG enzyme was ∼ 10 fold with a yield of 29%. The enzyme retained 98% activity in the presence of 15 % glycerol at 4 °C. The purified exo-PG using MNPs yielded a 10-12% increase in juice production as compare to without treated fruit juice. To the best of our knowledge, this is the first report of affinity purification of exo-PG enzyme, using engineered magnetic nanoparticles.
果胶酶水解果胶，占全球食品加工酶销售额的 25%。在本研究中，我们旨在使用半乳糖醛酸偶联磁纳米粒子（MNPs）纯化外切多聚半乳糖醛酸酶（Exo-PG），并研究其在果汁净化中的应用。在存在苹果果胶（1%）的情况下进行液体深层发酵，以促进黄曲霉产外切多聚半乳糖醛酸酶。在 30°C 和 pH 值为 5.0 的条件下，4 天后观察到最大的外切多聚半乳糖醛酸酶活性。SDS-PAGE 观察到纯化的外切多聚半乳糖醛酸酶只有一条单一的蛋白带（66 kDa）。酶的纯化倍数约为 10 倍，产率为 29%。在 4°C 下，15%甘油存在时，酶保留了 98%的活性。与未经处理的果汁相比，使用 MNPs 纯化的外切多聚半乳糖醛酸酶可使果汁产量增加 10-12%。据我们所知，这是首次使用工程化磁纳米粒子来亲和纯化外切多聚半乳糖醛酸酶的报道。
