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How to Engineer Organic Solvent Resistant Enzymes: Insights from Combined Molecular Dynamics and Directed Evolution Study
ChemCatChem ( IF 3.8 ) Pub Date : 2020-04-28 , DOI: 10.1002/cctc.202000422 Haiyang Cui 1 , Tom H. J. Stadtmüller 1 , Qianjia Jiang 1 , Karl‐Erich Jaeger 2 , Ulrich Schwaneberg 1, 3 , Mehdi D. Davari 1
ChemCatChem ( IF 3.8 ) Pub Date : 2020-04-28 , DOI: 10.1002/cctc.202000422 Haiyang Cui 1 , Tom H. J. Stadtmüller 1 , Qianjia Jiang 1 , Karl‐Erich Jaeger 2 , Ulrich Schwaneberg 1, 3 , Mehdi D. Davari 1
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Expanding synthetic capabilities to routinely employ enzymes in organic solvents (OSs) is a dream for protein engineers and synthetic chemists. Despite significant advances in the field of protein engineering, general and transferable design principles to improve the OS resistance of enzymes are poorly understood. Herein, we report a combined computational and directed evolution study of Bacillus subtlis lipase A (BSLA) in three OSs (i. e., 1,4‐dioxane, dimethyl sulfoxide, 2,2,2‐trifluoroethanol) to devise a rational strategy to guide engineering OS resistant enzymes. Molecular dynamics simulations showed that OSs reduce BSLA activity and resistance in OSs by (i) stripping off essential water molecules from the BLSA surface mainly through H‐bonds binding; and (ii) penetrating the substrate binding cleft leading to inhibition and conformational change. Interestingly, integration of computational results with “BSLA‐SSM” variant library (3439 variants; all natural diversity with amino acid exchange) revealed two complementary rational design strategies: (i) surface charge engineering, and (ii) substrate binding cleft engineering. These strategies are most likely applicable to stabilize other lipases and enzymes and assist experimentalists to design organic solvent resistant enzymes with reduced time and screening effort in lab experiments.
中文翻译：
如何设计耐有机溶剂酶：结合分子动力学和定向进化研究的见解
扩展合成能力以常规在有机溶剂（OSs）中使用酶是蛋白质工程师和合成化学家的梦想。尽管蛋白质工程领域取得了重大进展，但人们对提高酶对OS的抵抗力的通用和可移植设计原理知之甚少。在此，我们报告了芽孢杆菌的联合计算和定向进化研究脂肪酶A（BSLA）在三种OS（即1,4-二恶烷，二甲基亚砜，2,2,2-三氟乙醇）中制定合理的策略来指导工程OS抗性酶。分子动力学模拟表明，OS通过（i）主要通过H键结合从BLSA表面剥离必不可少的水分子来降低BSLA活性和OS中的抗性。（ii）穿透底物结合裂隙导致抑制和构象变化。有趣的是，将计算结果与“ BSLA-SSM”变体库（3439个变体；具有氨基酸交换的所有自然多样性）的整合揭示了两种互补的合理设计策略：（i）表面电荷工程和（ii）底物结合裂工程。
更新日期：2020-04-28
中文翻译：
如何设计耐有机溶剂酶：结合分子动力学和定向进化研究的见解
扩展合成能力以常规在有机溶剂（OSs）中使用酶是蛋白质工程师和合成化学家的梦想。尽管蛋白质工程领域取得了重大进展，但人们对提高酶对OS的抵抗力的通用和可移植设计原理知之甚少。在此，我们报告了芽孢杆菌的联合计算和定向进化研究脂肪酶A（BSLA）在三种OS（即1,4-二恶烷，二甲基亚砜，2,2,2-三氟乙醇）中制定合理的策略来指导工程OS抗性酶。分子动力学模拟表明，OS通过（i）主要通过H键结合从BLSA表面剥离必不可少的水分子来降低BSLA活性和OS中的抗性。（ii）穿透底物结合裂隙导致抑制和构象变化。有趣的是，将计算结果与“ BSLA-SSM”变体库（3439个变体；具有氨基酸交换的所有自然多样性）的整合揭示了两种互补的合理设计策略：（i）表面电荷工程和（ii）底物结合裂工程。
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