Competence Center for Biocatalysis, Institute of Chemistry and Biotechnology, School of Life Sciences and Facility Management, Zurich University of Applied Sciences, Einsiedlerstrasse 31, 8820, Wädenswil, Switzerland.
Molecular Biology, Institute of Chemistry and Biotechnology, School of Life Sciences and Facility Management, Zurich University of Applied Sciences, Einsiedlerstrasse 31, 8820, Wädenswil, Switzerland.
Chembiochem. 2019 Jun 14;20(12):1569-1577. doi: 10.1002/cbic.201800770. Epub 2019 May 8.
Many drug candidate molecules contain at least one chiral centre, and consequently, the development of biocatalytic strategies to complement existing metal- and organocatalytic approaches is of high interest. However, time is a critical factor in chemical process development, and thus, the introduction of biocatalytic steps, even if more suitable, is often prevented by the limited availability of off-the-shelf enzyme libraries. To expand the biocatalytic toolbox with additional ene reductases, we screened 19 bacterial strains for double bond reduction activity by using the model substrates cyclohexanone and carvone. Overall, we identified 47 genes coding for putative ene reductases. Remarkably, bioinformatic analysis of all genes and the biochemical characterization of four representative novel ene reductases led us to propose the existence of two new Old Yellow Enzyme subclasses, which we named OYE class III and class IV. Our results demonstrate that although, on a DNA level, each new OYE subclass features a distinct combination of sequence motifs previously known from the classical and the thermophilic-like group, their substrate scope more closely resembles the latter subclass.
许多候选药物分子都含有至少一个手性中心，因此，开发生物催化策略来补充现有的金属和有机催化方法是非常有意义的。然而，在化学工艺开发中，时间是一个关键因素，因此，即使生物催化步骤更合适，由于现成的酶库有限，通常也会阻止引入生物催化步骤。为了用额外的烯还原酶来扩展生物催化工具箱，我们使用模型底物环己酮和香芹酮筛选了 19 种细菌菌株的双键还原活性。总的来说，我们鉴定了 47 个编码潜在烯还原酶的基因。值得注意的是，对所有基因的生物信息学分析和对四个代表性新型烯还原酶的生化特征分析使我们提出了存在两种新的 Old Yellow Enzyme 亚类的假设，我们将其命名为 OYE 类 III 和类 IV。我们的结果表明，尽管在 DNA 水平上，每个新的 OYE 亚类都具有以前从经典和嗜热样亚类中已知的独特的序列基序组合，但它们的底物范围更类似于后者的亚类。
