Revealing Additional Stereocomplementary Pairs of Old Yellow Enzymes by Rational Transfer of Engineered Residues
Revealing Additional Stereocomplementary Pairs of Old Yellow Enzymes by Rational Transfer of Engineered Residues
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DOI：
10.1002/cbic.201600688
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发表时间：
2017-04-04
期刊：
影响因子：
3.2
通讯作者：
Hoebenreich, Sabrina
中科院分区：
文献类型：
--
作者：
Nett, Nathalie;Duewel, Sabine;Hoebenreich, Sabrina
每年都有大量的蛋白质工程和定向进化研究发表，增加了蛋白质工程师可以使用的知识。在这里，我们测试了一种蛋白质工程策略，该策略允许快速获得改进的生物催化剂，只需很少的筛选工作。从概念上讲，假设先前通过合理和随机方法鉴定的工程化残基在转移到家族成员时诱导类似的改善。在对来自Old Yellow Enzyme（OYE）家族的烯还原酶的应用中，用三种化合物测试了新产生的变体，揭示了更具立体互补性的OYE对，其具有有效的转换频率（高达660 h（-1））和优异的立体选择性（高达> 99%）。尽管系统预测OYE变体的绝对对映体选择性仍然是一个挑战，但“支架取样”被证实是蛋白质工程师收集策略的一个有前途的补充。
Every year numerous protein engineering and directed evolution studies are published, increasing the knowledge that could be used by protein engineers. Here we test a protein engineering strategy that allows quick access to improved biocatalysts with very little screening effort. Conceptually it is assumed that engineered residues previously identified by rational and random methods induce similar improvements when transferred to family members. In an application to ene-reductases from the Old Yellow Enzyme (OYE) family, the newly created variants were tested with three compounds, revealing more stereocomplementary OYE pairs with potent turnover frequencies (up to 660 h(-1)) and excellent stereoselectivities (up to > 99%). Although systematic prediction of absolute enantioselectivity of OYE variants remains a challenge, "scaffold sampling" was confirmed as a promising addition to protein engineers' collection of strategies.