Montgomery Sarah L, Pushpanath Ahir, Heath Rachel S, Marshall James R, Klemstein Ulrike, Galman James L, Woodlock David, Bisagni Serena, Taylor Christopher J, Mangas-Sanchez J, Ramsden J I, Dominguez Beatriz, Turner Nicholas J
School of Chemistry, University of Manchester, Manchester Institute of Biotechnology, 131 Princess Street, Manchester M1 7DN, UK.
Johnson Matthey, 28 Cambridge Science Park, Milton Road, Cambridge CB4 0FP, UK.
Sci Adv. 2020 May 22;6(21):eaay9320. doi: 10.1126/sciadv.aay9320. eCollection 2020 May.
Imine reductases (IREDs) have shown great potential as catalysts for the asymmetric synthesis of industrially relevant chiral amines, but a limited understanding of sequence activity relationships makes rational engineering challenging. Here, we describe the characterization of 80 putative and 15 previously described IREDs across 10 different transformations and confirm that reductive amination catalysis is not limited to any particular subgroup or sequence motif. Furthermore, we have identified another dehydrogenase subgroup with chemoselectivity for imine reduction. Enantioselectivities were determined for the reduction of the model substrate 2-phenylpiperideine, and the effect of changing the reaction conditions was also studied for the reductive aminations of 1-indanone, acetophenone, and 4-methoxyphenylacetone. We have performed sequence-structure analysis to help explain clusters in activity across a phylogenetic tree and to inform rational engineering, which, in one case, has conferred a change in chemoselectivity that had not been previously observed.
亚胺还原酶（IREDs）作为工业相关手性胺不对称合成的催化剂已显示出巨大潜力，但对序列活性关系的有限理解使得合理工程设计具有挑战性。在此，我们描述了对80种推定的和15种先前描述的IREDs在10种不同转化反应中的表征，并证实还原胺化催化不限于任何特定亚组或序列基序。此外，我们还鉴定出了另一个对亚胺还原具有化学选择性的脱氢酶亚组。测定了模型底物2-苯基哌啶的对映选择性，并研究了改变反应条件对茚满-1-酮、苯乙酮和4-甲氧基苯乙酮还原胺化反应的影响。我们进行了序列-结构分析，以帮助解释系统发育树中活性簇，并为合理工程设计提供信息，在一个案例中，这导致了以前未观察到的化学选择性变化。
