Structure-Oriented Engineering of Amidase: Modification of Twisted Access Tunnel for Efficient Synthesis of 2-Chloronicotinic Acid

酰胺酶 化学 产量(工程) 催化作用 位阻效应 组合化学 亲核细胞 吡啶 生物催化 水解 有机化学 反应机理 材料科学 冶金
作者
Zhe‐Ming Wu,Feng Xie,Wen Zheng,Changfeng Liu,Chao-Ping Lin,Ren‐Chao Zheng,Yu‐Guo Zheng
出处
期刊:ACS Catalysis [American Chemical Society]
卷期号:13 (13): 9078-9089 被引量:20
标识
DOI:10.1021/acscatal.3c01469
摘要

Amidases are robust biocatalysts for the industrial synthesis of carboxylic acids. Amidase-catalyzed hydrolysis of 2-chloronicotinamide is regarded as a promising route for 2-chloronicotinic acid production, which is an important building block for agrochemicals and pharmaceuticals. However, the o-substituted Cl of the pyridine ring caused a strong electronic effect and steric hindrance. Engineering access tunnels is one of the most efficient strategies to improve catalytic efficiency by adjusting the transfer efficiency of ligands. Herein, we engineered a twisted access tunnel of amidase and the mutant PaAmiM1A378V+V402L+L403V was achieved, which exhibited increased catalytic efficiency toward 2-chloronicotinamide. Tunnel analysis, molecular dynamics simulations, and quantum mechanical calculations revealed that the straightened tunnel significantly improved the transport efficiency and simultaneously facilitated substrate binding and nucleophilic attack. With the best mutant as a biocatalyst, 1330 mM product was accumulated within 2 h and a high space-time-yield of 2515 gproduct L–1 d–1 was achieved, which is the highest ever reported. These results lay the foundation for industrial production of 2-chloronicotinic acid and give important guidance for engineering amidases into versatile biocatalysts.
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
PDF的下载单位、IP信息已删除 (2025-6-4)

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
周老八发布了新的文献求助10
刚刚
刚刚
叶泽完成签到,获得积分10
刚刚
lx发布了新的文献求助10
1秒前
栗子发布了新的文献求助10
1秒前
1秒前
我是老大应助科研通管家采纳,获得10
1秒前
桐桐应助科研通管家采纳,获得10
1秒前
1秒前
CodeCraft应助科研通管家采纳,获得10
1秒前
2秒前
2秒前
大模型应助科研通管家采纳,获得10
2秒前
2秒前
2秒前
丘比特应助円桑采纳,获得10
2秒前
64658应助清秀的怀蕊采纳,获得10
2秒前
xingxingxing发布了新的文献求助10
3秒前
彭于彦祖应助微醺小王采纳,获得30
3秒前
4秒前
4秒前
fyl发布了新的文献求助10
4秒前
周老八完成签到,获得积分10
5秒前
5秒前
even发布了新的文献求助10
5秒前
7秒前
有魅力向珊完成签到,获得积分20
7秒前
Lucas应助咩咩采纳,获得10
7秒前
8秒前
徐逊发布了新的文献求助10
8秒前
李健应助潇洒的凌兰采纳,获得10
8秒前
whisper发布了新的文献求助30
8秒前
9秒前
李健的小迷弟应助刘66采纳,获得10
9秒前
yyy完成签到,获得积分20
10秒前
面包发布了新的文献求助30
10秒前
爱吃冻梨完成签到,获得积分10
10秒前
ranranran完成签到,获得积分10
11秒前
Joan_89发布了新的文献求助20
11秒前
周星星同学完成签到,获得积分10
11秒前
高分求助中
A new approach to the extrapolation of accelerated life test data 1000
Cognitive Neuroscience: The Biology of the Mind 1000
Technical Brochure TB 814: LPIT applications in HV gas insulated switchgear 1000
ACSM’s Guidelines for Exercise Testing and Prescription, 12th edition 500
Picture Books with Same-sex Parented Families: Unintentional Censorship 500
Nucleophilic substitution in azasydnone-modified dinitroanisoles 500
不知道标题是什么 500
热门求助领域 (近24小时)
化学 材料科学 医学 生物 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 遗传学 基因 物理化学 催化作用 冶金 细胞生物学 免疫学
热门帖子
关注 科研通微信公众号,转发送积分 3969383
求助须知:如何正确求助?哪些是违规求助? 3514211
关于积分的说明 11172730
捐赠科研通 3249476
什么是DOI,文献DOI怎么找? 1794909
邀请新用户注册赠送积分活动 875441
科研通“疑难数据库(出版商)”最低求助积分说明 804827