相关实验视频

Updated: Jun 21, 2025

Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins

Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins

Published on: June 20, 2019

对一个甲酸乙酸的催化机制的分子洞察力

Ning Wang¹, Nan Zhang², Mei-Ling Sun³

¹MOE Key Laboratory of Evolution and Marine Biodiversity, Frontiers Science Center for Deep Ocean Multispheres and Earth System & College of Marine Life Sciences, Ocean University of China, Qingdao, China; State Key Laboratory of Microbial Technology, Marine Biotechnology Research Center, Shandong University, Qingdao, China; Joint Research Center for Marine Microbial Science and Technology, Shandong University and Ocean University of China, Qingdao, China.

Journal of hazardous materials

|July 16, 2024

概括

这项研究特征HylD1,一个家族IV雌激酶,揭示其结构和催化机制降解甲酸 (PAE). 这为开发高效的PAE降解酶提供了基础.

科学领域:

生物化学生化学
环境科学环境科学
酶学是一种酶学.

背景情况:

酸盐 (PAE) 是广泛存在的环境污染物.
乙酶家族IV是PAE降解的关键酶,但它们的结构基本上是未知的.
甲基甲甲酸 (DEP) 和甲基甲甲酸 (DMP) 是首要的PAE污染物.

研究的目的:

为了表征PAE酶HylD1,一个家族IV酶.
为了确定HylD1的晶体结构及其与基质模拟的复合物.
阐明催化机制并确定HylD1.1的基质结合残留物.

主要方法:

重组HylD1的表达和特征.
确定HylD1晶体结构 (apo和复杂).
生物化学测定和机理学研究.

主要成果:

HylD1在广泛的pH范围内作为二聚体起作用.
晶体结构揭示了基质结合残留物和活性部位特征.
提出了一种涉及Ser140-Asp231-His261三合体的催化机制.
hylD1 基因在各种环境中广泛分布.

结论:

HylD1是一种有效的PAE酶,具有明确的结构和催化机制.
了解HylD1为PAE水解提供了洞察力.
这项工作支持用于工业应用的新型PAE降解酶的合理设计.

关键词:

催化机制是一种催化机制.晶体结构晶体结构甲基甲基甲酸二甲基甲酸二甲基甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸甲基二甲甲酸二甲基甲酸二甲基甲酸二甲基甲酸二甲酸二甲基甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸二甲酸甲酸 Ester 氨酸酸酶.

更多相关视频

Defining Substrate Specificities for Lipase and Phospholipase Candidates

Defining Substrate Specificities for Lipase and Phospholipase Candidates

Published on: November 23, 2016

Phthalic Acid Ester-Binding DNA Aptamer Selection, Characterization, and Application to an Electrochemical Aptasensor

Phthalic Acid Ester-Binding DNA Aptamer Selection, Characterization, and Application to an Electrochemical Aptasensor

Published on: March 21, 2018

相关概念视频

Esters to Carboxylic Acids: Acid-Catalyzed Hydrolysis

Esters to Carboxylic Acids: Acid-Catalyzed Hydrolysis

Hydrolysis of esters under acidic conditions proceeds through a nucleophilic acyl substitution. In the presence of excess water, the reaction proceeds in a reversible manner, forming carboxylic acids and alcohols.
During hydrolysis, the ester is first activated towards nucleophilic attack through the protonation of the carboxyl oxygen atom by the acid catalyst. The protonation makes the ester carbonyl carbon more electrophilic. In the next step, water acts as a nucleophile and adds to the...

Acid Halides to Esters: Alcoholysis

Acid Halides to Esters: Alcoholysis

Alcoholysis is a nucleophilic acyl substitution reaction in which an alcohol functions as a nucleophile. Acid halides react with alcohol to produce esters. The mechanism proceeds in three steps:

Regioselectivity and Stereochemistry of Acid-Catalyzed Hydration

Regioselectivity and Stereochemistry of Acid-Catalyzed Hydration

The rate of acid-catalyzed hydration of alkenes depends on the alkene's structure, as the presence of alkyl substituents at the double bond can significantly influence the rate.

Carboxylic Acids to Esters: Acid-Catalyzed (Fischer) Esterification Mechanism

Carboxylic Acids to Esters: Acid-Catalyzed (Fischer) Esterification Mechanism

Carboxylic acids react with alcohols to yield esters via an acid-catalyzed condensation reaction called Fischer esterification. This is a nucleophilic acyl substitution reaction that proceeds via a tetrahedral intermediate, where a water molecule is eliminated as the leaving group.

E2 Reaction: Kinetics and Mechanism

E2 Reaction: Kinetics and Mechanism

SN2 substitutions and E2 eliminations of alkyl halides proceed via a concerted pathway. While the nucleophile attacks the alpha carbon in SN2 reactions, it functions as a strong base and abstracts a beta hydrogen in the E2 mechanism. The rate-limiting transition state in E2 elimination reactions is characterized by partially broken carbon–hydrogen and carbon–halogen bonds and a partially formed pi bond between the alpha and beta carbons. The beta hydrogen and halide are eliminated...

Esters to β-Ketoesters: Claisen Condensation Mechanism

Esters to β-Ketoesters: Claisen Condensation Mechanism

Regular Claisen condensation involves the synthesis of β-ketoesters by combining identical ester molecules bearing two α hydrogens in the presence of an alkoxide base. The reaction commences with the deprotonation of the acidic α hydrogen by the base to form a resonance stabilized ester enolate. This nucleophilic ion then attacks the carbonyl center of another ester molecule to generate a tetrahedral alkoxide intermediate. Next, the expulsion of the alkoxide group from the...