基于脯氨酰内肽酶完美构象的能量优化策略，提高催化效率。
An Energy Optimization Strategy Based on the Perfect Conformation of Prolyl Endopeptidase for Improving Catalytic Efficiency.
机构信息
Shanghai Engineering Research Center of Molecular Therapeutics & New Drug Development, Shanghai Key Laboratory of Green Chemistry & Chemical Process, School of Chemistry and Molecular Engineering, East China Normal University, Shanghai 200062, China.
School of Biotechnology, East China University of Science and Technology, Shanghai 200237, China.
出版信息
J Agric Food Chem. 2020 May 6;68(18):5129-5137. doi: 10.1021/acs.jafc.0c00731. Epub 2020 Apr 27.
Prolyl endopeptidases (PEPs) hydrolyze proteins to yield bioactive peptides and are effective in the treatment of celiac disease. However, the catalytic efficiency of PEPs still has the potential to be improved, which could further strengthen their industrial and therapeutic applications. Herein, a novel rational design strategy based on a "near-attack conformation" of the catalytic state of PEP was adopted. Constrained dynamic simulations were applied, followed by the virtual screening of potentially favorable mutants according to their binding free energy. We redesigned PEP with high-temperature activity/stability, a wide range of pH stabilities, and high proline specificity. As a result, the value of two PEP mutants (I462W and Q560Y) increased by 208.2 and 150.1%, respectively, and the / increased by 32.7 and 6.3%, respectively. These data revealed that the PEP mutants had improved catalytic efficiency and that our strategy can be applied for enzyme engineering.
脯氨酰内肽酶（PEPs）能够水解蛋白质生成具有生物活性的肽，可有效治疗乳糜泻。然而，PEPs 的催化效率仍有提升空间，这将进一步增强其在工业和治疗领域的应用。本研究采用一种基于 PEP 催化态“近攻击构象”的新型理性设计策略。通过约束动态模拟，根据结合自由能对潜在有利的突变体进行虚拟筛选。我们重新设计了具有高温活性/稳定性、宽 pH 稳定性和高脯氨酸特异性的 PEP。结果，两个 PEP 突变体（I462W 和 Q560Y）的 值分别提高了 208.2%和 150.1%， / 分别提高了 32.7%和 6.3%。这些数据表明突变体具有更高的催化效率，且我们的策略可应用于酶工程。
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