Yukselen Onur, Timucin Emel, Sezerman Ugur
Biological Sciences and Bioengineering, Faculty of Engineering and Natural Sciences, Sabanci University, Istanbul, 34956, Turkey.
Biostatistics and Medical Informatics, School of Medicine, Acibadem University, Istanbul, 34752, Turkey.
J Mol Recognit. 2016 Oct;29(10):466-75. doi: 10.1002/jmr.2545. Epub 2016 Apr 14.
Lipases are important biocatalysts owing to their ability to catalyze diverse reactions with exceptional substrate specificities. A combined docking and molecular dynamics (MD) approach was applied to study the chain-length selectivity of Bacillus thermocatenulatus lipase (BTL2) towards its natural substrates (triacylglycerols). A scoring function including electrostatic, van der Waals (vdW) and desolvation energies along with conformational entropy was developed to predict the impact of mutation. The native BTL2 and its 6 mutants (F17A, V175A, V175F, D176F, T178V and I320F) were experimentally analyzed to determine their specific activities towards tributyrin (C4) or tricaprylin (C8), which were used to test our approach. Our scoring methodology predicted the chain-length selectivity of BTL2 with 85.7% (6/7) accuracy with a positive correlation between the calculated scores and the experimental activity values (r = 0.82, p = 0.0004). Additionally, the impact of mutation on activity was predicted with 75% (9/12) accuracy. The described study represents a fast and reliable approach to accurately predict the effect of mutations on the activity and selectivity of lipases and also of other enzymes. Copyright © 2016 John Wiley & Sons, Ltd.
脂肪酶是重要的生物催化剂，因为它们能够以特殊的底物特异性催化多种反应。采用对接与分子动力学（MD）相结合的方法研究了嗜热链霉菌脂肪酶（BTL2）对其天然底物（三酰甘油）的链长选择性。开发了一种包含静电、范德华（vdW）和去溶剂化能以及构象熵的评分函数，以预测突变的影响。对天然BTL2及其6个突变体（F17A、V175A、V175F、D176F、T178V和I320F）进行了实验分析，以确定它们对三丁酸甘油酯（C4）或三辛酸甘油酯（C8）的比活性，这些用于测试我们的方法。我们的评分方法预测BTL2链长选择性的准确率为85.7%（6/7），计算得分与实验活性值之间呈正相关（r = 0.82，p = 0.0004）。此外，预测突变对活性影响的准确率为75%（9/12）。所描述的研究代表了一种快速且可靠的方法，可准确预测突变对脂肪酶以及其他酶的活性和选择性的影响。版权所有© 2016约翰威立父子有限公司。
