Center of Plasma Nano-interface Engineering, Kyushu University, Fukuoka, Japan.
Department of Biochemistry, College of Life Science & Biotechnology, Yonsei University, Seoul 120-749, Republic of Korea.
Int J Biol Macromol. 2020 Jul 15;155:439-446. doi: 10.1016/j.ijbiomac.2020.03.175. Epub 2020 Mar 24.
Ionic liquids (ILs) are known to provide stability to biomolecules. ILs are also widely used in the fields of chemical engineering, biological engineering, chemistry, and biochemistry because they facilitate enzyme catalyzed reactions and enhance their conversion rate. In this work, we have evaluated the influence of alkyl chain substitution of ammonium ILs such as diethylammonium dihydrogen phosphate (DEAP) and triethylammonium hydrogen phosphate (TEAP) for the stability and activity of the tobacco etch virus (TEV) protease. Further, we performed molecular dynamics (MD) simulations to calculate the RMSD (root mean square deviation) for TEV and TEV + ILs. Experimental and simulations results show that TEV is more stable in the presence of TEAP than DEAP. Whereas, TEV protease activity for the cleavage of fusion proteins is preserved in the presence of DEAP while lost in the presence of TEAP. Hence, DEAP IL can serve as alternative solvents for the stability of the TEV protease with preserved activity. To the best of our knowledge, this is first study to show that ILs can stabilize and maintain the TEV protease cleavage activity.
离子液体 (ILs) 已知能为生物分子提供稳定性。由于它们有利于酶催化反应并提高其转化率，因此 ILs 在化学工程、生物工程、化学和生物化学等领域也得到了广泛应用。在这项工作中，我们评估了铵离子 ILs（如二乙基磷酸二氢铵 (DEAP) 和三乙基磷酸氢铵 (TEAP)）的烷基链取代对烟草蚀纹病毒 (TEV) 蛋白酶稳定性和活性的影响。此外，我们进行了分子动力学 (MD) 模拟，以计算 TEV 和 TEV+ILs 的 RMSD（均方根偏差）。实验和模拟结果表明，在 TEAP 存在下，TEV 比 DEAP 更稳定。然而，在 DEAP 存在下，TEV 蛋白酶对融合蛋白的切割活性得以保留，而在 TEAP 存在下则丧失。因此，DEAP IL 可以作为替代溶剂，保持 TEV 蛋白酶的稳定性和活性。据我们所知，这是首次表明 ILs 可以稳定和维持 TEV 蛋白酶切割活性的研究。
