Lutz Stefan, Iamurri Samantha M
Department of Chemistry, Emory University, 1515 Dickey Drive, Atlanta, GA, 30322, USA.
Methods Mol Biol. 2018;1685:1-12. doi: 10.1007/978-1-4939-7366-8_1.
The last decade has seen a dramatic increase in the utilization of enzymes as green and sustainable (bio)catalysts in pharmaceutical and industrial applications. This trend has to a significant degree been fueled by advances in scientists' and engineers' ability to customize native enzymes by protein engineering. A review of the literature quickly reveals the tremendous success of this approach; protein engineering has generated enzyme variants with improved catalytic activity, broadened or altered substrate specificity, as well as raised or reversed stereoselectivity. Enzymes have been tailored to retain activity at elevated temperatures and to function in the presence of organic solvents, salts and pH values far from physiological conditions. However, readers unfamiliar with the field will soon encounter the confusingly large number of experimental techniques that have been employed to accomplish these engineering feats. Herein, we use history to guide a brief overview of the major strategies for protein engineering-past, present, and future.
在过去十年中，酶作为绿色且可持续的（生物）催化剂在制药和工业应用中的使用量急剧增加。科学家和工程师通过蛋白质工程定制天然酶的能力取得了进展，这在很大程度上推动了这一趋势。对文献的回顾很快就能发现这种方法取得的巨大成功；蛋白质工程已产生了具有改进催化活性、拓宽或改变底物特异性以及提高或反转立体选择性的酶变体。酶经过改造后能够在高温下保持活性，并能在远离生理条件的有机溶剂、盐和pH值环境中发挥作用。然而，不熟悉该领域的读者很快就会遇到为实现这些工程壮举而采用的数量多得令人困惑的实验技术。在此，我们借助历史来简要概述蛋白质工程的主要策略——过去、现在和未来。
