检索结果-南通市图书馆

Enhancing the activity and succinyl-CoA specificity of 3-ketoacyl-CoA thiolase Tfu_0875 through rational binding pocket engineering

引用

Synthetic and Systems Biotechnology 2024年第3期9卷 558-568页

作者： Lixia Liu Shuang Liu Xiangyang Hu Shenghu Zhou Yu Deng National Engineering Research Center of Cereal Fermentation and Food Biomanufacturing Jiangnan University1800 Lihu RoadWuxiJiangsu214122China Jiangsu Provincial Research Center for Bioactive Product Processing Technology Jiangnan University1800 Lihu RoadWuxiJiangsu214122China

The 3-ketoacyl-CoA thiolase is the rate-limiting enzyme for linear dicarboxylic acids ***,the promiscuous substrate specificity and suboptimal catalytic performance have restricted its *** we present both biochemical and structural analyses of a high-efficiency 3-ketoacyl-CoA thiolase Tfu_***,Tfu_0875 displayed heightened activity and substrate specificity for succinyl-CoA,a key precursor in adipic acid *** enhance its performance,a deep learning approach(DLKcat)was employed to identify effective mutants,and a computational strategy,known as the greedy accumulated strategy for protein engineering(grape),was used to accumulate these effective *** the mutants,Tfu_0875N249W/L163H/E217L exhibited the highest specific activity(320%of wild-type Tfu_0875),the greatest catalytic efficiency(kcat/KM=1.00 min-1mM-1),the highest succinyl-CoA specificity(KM=0.59 mM,28.1%of Tfu_0875)and dramatically reduced substrate binding energy(-30.25 kcal mol^(-1)v.s.-15.94 kcal mol^(-1)).A structural comparison between Tfu_0875N249W/L163H/E217L and the wild type Tfu_0875 revealed that the increased interaction between the enzyme and succinyl-CoA was the primary reason for the enhanced enzyme *** interaction facilitated rapid substrate anchoring and ***,a reduced binding pocket volume improved substrate specificity by enhancing the complementarity between the binding pocket and the substrate in stereo ***,our rationally designed mutant,Tfu_0875N249W/L163H/E217L,increased the adipic acid titer by 1.35-fold compared to the wild type Tfu_0875 in shake *** demonstrated enzymatic methods provide a promising enzyme variant for the adipic acid *** above effective substrate binding pocket engineering strategy can be beneficial for the production of other industrially competitive biobased chemicals when be applied to other thiolases.

关键词： Adipic acid 3-Ketoacyl-CoA thiolase Claisen condensation Deep learning grape strategy

来源：

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

欢迎您,

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

请选择保存的检索档案：

请选择收藏分类：

通借通还

欢迎您,

建议与咨询 留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

请选择保存的检索档案： 新增检索档案 确定 取消

请选择收藏分类： 新增自定义分类 确定 取消

通借通还

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

请选择保存的检索档案：

请选择收藏分类：