Zheng Ling, Guo Zilong, Cao Shengsheng, Zhu Benwei
College of Food Science and Light Industry, Nanjing Tech University, Nanjing, 211816, China.
Bioresour Bioprocess. 2021 Dec 4;8(1):121. doi: 10.1186/s40643-021-00475-2.
The cold-active pectate lyases have drawn increasing attention in food and biotechnological applications due to their ability to retain high catalytic efficiency under lower temperatures, which could be helpful for energy saving, cost reduction and flavor preservation. Herein, a new cold-tolerant pectate lyase (ErPelPL1) gene from Echinicola rosea was cloned and heterologously expressed in Escherichia coli. Interestingly, ErPelPL1 retained high catalytic activity even at a low temperature (4 °C). ErPelPL1 exhibited optimal activity at 35 ℃, pH 8.0 with 1 mM of Ca. It showed high specific activity towards polygalacturonic acid (34.7 U/mg) and sodium polygalacturonate (59.3 U/mg). The combined thin-layer chromatography (TLC), fast protein liquid chromatography (FPLC) and electrospray ionization mass spectrometry (ESI-MS) results indicated that ErPelPL1 endolytically degraded pectic substances into the oligosaccharides with degrees of depolymerization (Dps) of 1-6. In conclusion, this study mainly conducted biochemical characterization and product analysis of a cold-tolerant pectate lyase. Therefore, it provides a promising enzyme candidate for food and biotechnological applications.
由于冷活性果胶酸裂解酶在较低温度下仍能保持较高的催化效率，这有助于节能、降低成本和保存风味，因此在食品和生物技术应用中受到越来越多的关注。在此，从玫瑰海栖热袍菌中克隆了一种新的耐冷果胶酸裂解酶（ErPelPL1）基因，并在大肠杆菌中进行了异源表达。有趣的是，即使在低温（4℃）下，ErPelPL1仍保持较高的催化活性。ErPelPL1在35℃、pH 8.0和1 mM Ca条件下表现出最佳活性。它对聚半乳糖醛酸（34.7 U/mg）和聚半乳糖醛酸钠（59.3 U/mg）表现出较高的比活性。薄层色谱（TLC）、快速蛋白质液相色谱（FPLC）和电喷雾电离质谱（ESI-MS）的联合结果表明，ErPelPL1将果胶物质内切降解为解聚度（Dps）为1-6的寡糖。总之，本研究主要对一种耐冷果胶酸裂解酶进行了生化特性分析和产物分析。因此，它为食品和生物技术应用提供了一种有前景的酶候选物。
