Türkoğlu Emir Alper, Taştekil Ilgaz, Özbek Sarica Pemra
Department of Pharmaceutical Biotechnology, Faculty of Pharmacy University of Health Sciences Turkey İstanbul Türkiye.
Department of Bioengineering Institute of Pure and Applied Sciences, Marmara University İstanbul Türkiye.
Food Sci Nutr. 2024 Jul 22;12(10):7392-7404. doi: 10.1002/fsn3.4315. eCollection 2024 Oct.
Lysozyme (L) is an important enzyme in medicine and industry. Tannic acid (TA) is used in brewing, wine industry, and as a food flavor enhancer. In nutritional and food science, L interacts with TA, notably in wine and saliva. This study aimed to investigate the binding interaction between L and TA using surface plasmon resonance, molecular docking, and molecular dynamics simulation. Chicken egg white lysozyme (CEWL) was applied as a model protein. Tri-N-acetylchitotriose (NAG), the known inhibitor of CEWL, was used in the redocking experiments to determine the precise binding location within the complex. The average binding energies obtained from docking NAG and tannic acid to the target structure of CEWL were found to be -6.46 ± 0.05 kcal/mol and -7.52 ± 0.39 kcal/mol, respectively. The binding free energy of the CEWL-TA complex was then calculated as -27.61 kcal/mol by MMPBSA based on MD simulation trajectories. The observed interactions between the ligands and the lysozyme structure were mainly driven by hydrophobic, van der Waals, and H-bond interactions formed by the active site residues. MD simulations showed consistent and satisfactory binding distances between CEWL and TA throughout the analysis. SPR analysis was performed using 1X PBS buffer (pH 7.4) as coupling and running buffers, 30 μL/min as flow rate, and 2.5 mg/mL CEWL. Serial concentrations of TA (20-150 μM) were injected through immobilized CEWL, and the value of CEWL-TA binding was obtained as 4.17 × 10 M. This study could enhance existing literature and pave the way for future research in food science and oral biology.
溶菌酶（L）在医学和工业中是一种重要的酶。单宁酸（TA）用于酿造、葡萄酒工业以及作为食品风味增强剂。在营养和食品科学领域，L与TA相互作用，尤其是在葡萄酒和唾液中。本研究旨在利用表面等离子体共振、分子对接和分子动力学模拟来研究L与TA之间的结合相互作用。以鸡蛋白溶菌酶（CEWL）作为模型蛋白。在重新对接实验中使用了CEWL的已知抑制剂三 - N - 乙酰壳三糖（NAG）来确定复合物内的精确结合位置。将NAG和单宁酸对接至CEWL的靶结构所获得的平均结合能分别为 -6.46±0.05 kcal/mol和 -7.52±0.39 kcal/mol。然后基于分子动力学模拟轨迹通过MMPBSA计算出CEWL - TA复合物的结合自由能为 -27.61 kcal/mol。观察到配体与溶菌酶结构之间的相互作用主要由活性位点残基形成的疏水、范德华和氢键相互作用驱动。分子动力学模拟在整个分析过程中显示CEWL与TA之间具有一致且令人满意的结合距离。表面等离子体共振分析使用1X PBS缓冲液（pH 7.4）作为偶联和运行缓冲液，流速为30 μL/min，CEWL浓度为2.5 mg/mL。将系列浓度的TA（20 - 150 μM）注入固定化的CEWL，得到CEWL - TA结合的 值为4.17×10 M。本研究可以丰富现有文献，并为食品科学和口腔生物学的未来研究铺平道路。
