Structural Insight into Enantioselective Inversion of an Alcohol Dehydrogenase Reveals a “Polar Gate” in Stereorecognition of Diaryl Ketones

化学 对映选择合成 反演(地质) 醇脱氢酶 极地的 组合化学 立体化学 有机化学 催化作用 天文 生物 构造盆地 物理 古生物学
作者
Jieyu Zhou,Yue Wang,Guochao Xu,Lian Wu,Ruizhi Han,Ulrich Schwaneberg,Yijian Rao,Yi‐Lei Zhao,Jiahai Zhou,Ye Ni
出处
期刊:Journal of the American Chemical Society [American Chemical Society]
卷期号:140 (39): 12645-12654 被引量:101
标识
DOI:10.1021/jacs.8b08640
摘要

Diaryl ketones are important building blocks for synthesizing pharmaceuticals and are generally regarded as "difficult-to-reduce" ketones due to the large steric hindrance of their two bulky aromatic side chains. Alcohol dehydrogenase from Kluyveromyces polyspora (KpADH) has been identified as a robust biocatalyst due to its high conversion of diaryl ketone substrate (4-chlorophenyl)(pyridine-2-yl)ketone (CPMK) with a moderate R-selectivity of 82% ee. To modulate the stereoselectivity of KpADH, a "polarity scanning" strategy was proposed, in which six key residues inside and at the entrance of the substrate binding pocket were identified. After iterative combinatorial mutagenesis, variants Mu-R2 and Mu-S5 with enhanced (99.2% ee, R) and inverted (97.8% ee, S) stereoselectivity were obtained. The crystal structures of KpADH and two mutants in complex with NADPH were resolved to elucidate the evolution of enantioselective inversion. Based on MD simulation, Mu-R2–CPMKProR and Mu-S5–CPMKProS were more favorable in the formation of prereaction states. Interestingly, a quadrilateral plane formed by α-carbons of four residues (N136, V161, C237, and G214) was identified at the entrance of the substrate binding pocket of Mu-S5; this plane acts as a "polar gate" for substrates. Due to the discrepancy in charge characteristics between chlorophenyl and pyridine substituents, the pro-S orientation of CPMK is defined when it passes through the "polar gate" in Mu-S5, whereas the similar plane in wild-type is blocked by several aromatic residues. Our result paves the way for engineering stereocomplementary ADH toward bulky diaryl ketones and provides structural insight into the mechanism of stereoselective inversion.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
大幅提高文件上传限制,最高150M (2024-4-1)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
漫漫完成签到,获得积分10
刚刚
111112完成签到,获得积分10
刚刚
笑南发布了新的文献求助10
刚刚
Hahawang发布了新的文献求助10
2秒前
Jasper应助到家了采纳,获得10
2秒前
李爱国应助睡睡采纳,获得10
2秒前
土豪的山蝶完成签到,获得积分10
2秒前
向前完成签到,获得积分10
3秒前
3秒前
3秒前
共享精神应助LFJ采纳,获得10
4秒前
5秒前
打打应助cz采纳,获得10
6秒前
哈哈哈发布了新的文献求助10
7秒前
7秒前
7秒前
Hahawang完成签到,获得积分10
7秒前
Gstar完成签到,获得积分10
8秒前
高高浩然发布了新的文献求助10
8秒前
8R60d8应助哈哈哈采纳,获得10
8秒前
王加冕发布了新的文献求助10
11秒前
机智乐菱完成签到,获得积分10
13秒前
张张发布了新的文献求助10
14秒前
Willing发布了新的文献求助10
14秒前
江湖郎中发布了新的文献求助10
14秒前
ty120完成签到,获得积分10
15秒前
明理往事完成签到 ,获得积分10
15秒前
16秒前
科研通AI2S应助YOUNG-M采纳,获得10
17秒前
王建平完成签到 ,获得积分10
18秒前
大模型应助实验室的亡灵采纳,获得10
19秒前
19秒前
20秒前
chenxin7271发布了新的文献求助10
22秒前
Willing完成签到,获得积分10
22秒前
青枣不甜完成签到,获得积分10
22秒前
22秒前
24秒前
机智乐菱发布了新的文献求助10
25秒前
善学以致用应助Cwx2020采纳,获得10
25秒前
高分求助中
Sustainability in Tides Chemistry 2800
The Young builders of New china : the visit of the delegation of the WFDY to the Chinese People's Republic 1000
юрские динозавры восточного забайкалья 800
English Wealden Fossils 700
Foreign Policy of the French Second Empire: A Bibliography 500
Chen Hansheng: China’s Last Romantic Revolutionary 500
Classics in Total Synthesis IV 400
热门求助领域 (近24小时)
化学 医学 生物 材料科学 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 基因 遗传学 催化作用 物理化学 免疫学 量子力学 细胞生物学
热门帖子
关注 科研通微信公众号,转发送积分 3146046
求助须知:如何正确求助?哪些是违规求助? 2797450
关于积分的说明 7824222
捐赠科研通 2453810
什么是DOI,文献DOI怎么找? 1305876
科研通“疑难数据库(出版商)”最低求助积分说明 627593
版权声明 601491