相关实验视频

Updated: Jun 15, 2025

Measuring Enzymatic Stability by Isothermal Titration Calorimetry

Measuring Enzymatic Stability by Isothermal Titration Calorimetry

Published on: March 26, 2019

环动力学工程超越了活性稳定性权衡的障碍,并提高了酶的热稳定性

Wenya Chong¹, Zihan Zhang¹, Zhongyu Li²

¹State Key Laboratory of Materials-Oriented Chemical Engineering, School of Pharmaceutical Sciences, Nanjing Tech university, Nanjing 210009, Jiangsu Province, People's Republic of China.

International journal of biological macromolecules

|August 24, 2024

概括

工程师开发了一种"环动力学工程" (HLoD) 策略,以提高酶的热稳定性. 这种方法提高了Candida Antarctica脂酶B的稳定性七倍,同时保持其活性,克服了典型的活性-稳定性权衡.

科学领域:

酶工程是什么?酶工程是什么?
蛋白质的结构动态蛋白质的结构动态
生物催化剂是一种生物催化剂.

背景情况:

提高酶的热稳定性往往会降低活性,因为关键区域的灵活性下降.
了解循环动态对于开发有效的酶稳定策略至关重要.
酶的结构动态与它们的功能密切相关.

研究的目的:

提出一种新的策略,即"环动力学工程" (HLoD),用于同时提高酶的热稳定性和活性.
调查修改特定"子循环"对Candida Antarctica B.脂酶性质的影响.
探索循环动力学,分子间相互作用和酶稳定性之间的关系.

主要方法:

设计和构建了一个小型突变库,针对Candida Antarctica lipase B"圈"中的五个关键残留物.
系统地研究了产生的突变体的热稳定性和活性.
采用分子动力学模拟来分析野生类型和突变酶的结构和动态变化.

主要成果:

与野生型 (WT) 相比,一个五点多重突变 (M1) 在60°C的温度稳定性增加了7.0倍.
突变M1保持了与WT相似的活性,有效地克服了活动稳定性权衡.
分子动力学模拟表明,通过盐桥和键的调节环动力学,减少了关键区域的过度灵活性,提高了热稳定性.

结论:

HLoD策略是一种强大的方法,可以同时提高酶的热稳定性和活性.
改进灵活区域的动态,为设计高度稳定和活性酶提供了通用策略.
这项研究为酶中的结构-动力学-功能关系提供了新的见解.

关键词:

活动稳定性的权衡.计算机辅助设计是指计算机辅助设计.环动力学工程环动力学工程循环的动态循环的动态热稳定性热稳定性

更多相关视频

A New Screening Method for the Directed Evolution of Thermostable Bacteriolytic Enzymes

A New Screening Method for the Directed Evolution of Thermostable Bacteriolytic Enzymes

Published on: November 7, 2012

Unraveling Entropic Rate Acceleration Induced by Solvent Dynamics in Membrane Enzymes

Unraveling Entropic Rate Acceleration Induced by Solvent Dynamics in Membrane Enzymes

Published on: January 16, 2016

相关概念视频

Introduction to Mechanisms of Enzyme Catalysis

Introduction to Mechanisms of Enzyme Catalysis

For many years, scientists thought that enzyme-substrate binding took place in a simple "lock-and-key" fashion. This model stated that the enzyme and substrate fit together perfectly in one instantaneous step. However, current research supports a more refined view scientists call induced fit. The induced-fit model expands upon the lock-and-key model by describing a more dynamic interaction between enzyme and substrate. As the enzyme and substrate come together, their interaction causes...

Enzymes

Enzymes

Inside living organisms, enzymes act as catalysts for many biochemical reactions involved in cellular metabolism. The role of enzymes is to reduce the activation energies of biochemical reactions by forming complexes with its substrates. The lowering of activation energies favor an increase in the rates of biochemical reactions.
Enzyme deficiencies can often translate into life-threatening diseases. For example, a genetic abnormality resulting in the deficiency of the enzyme G6PD...

Catalytically Perfect Enzymes

Catalytically Perfect Enzymes

The theory of catalytically perfect enzymes was first proposed by W.J. Albery and J. R. Knowles in 1976. These enzymes catalyze biochemical reactions at high-speed. Their catalytic efficiency values range from 108-109 M-1s-1. These enzymes are also called 'diffusion-controlled' as the only rate-limiting step in the catalysis is that of the substrate diffusion into the active site. Examples include triose phosphate isomerase, fumarase, and superoxide dismutase.

Most enzymes...

Induced-fit Model

Induced-fit Model

Most chemical reactions in cells require enzymes—biological catalysts that speed up the reaction without being consumed or permanently changed. They reduce the activation energy needed to convert the reactants into products. Enzymes are proteins, that usually work by binding to a substrate—a reactant molecule that they act upon.
Enzymes exhibit substrate specificity, meaning that they can only bind to certain substrates. This is mainly determined by the shape and chemical...

Enzyme Kinetics

Enzyme Kinetics

Enzymes speed up reactions by lowering the activation energy of the reactants. The speed at which the enzyme turns reactants into products is called the rate of reaction. Several factors impact the rate of reaction, including the number of available reactants. Enzyme kinetics is the study of how an enzyme changes the rate of a reaction.
Scientists typically study enzyme kinetics with a fixed amount of enzyme in the controlled environment of a test tube. When more reactant, or substrate, is...

Allosteric Regulation

Allosteric Regulation

Allosteric regulation of enzymes occurs when the binding of an effector molecule to a site that is different from the active site causes a change in the enzymatic activity. This alternate site is called an allosteric site, and an enzyme can contain more than one of these sites. Allosteric regulation can either be positive or negative, resulting in an increase or decrease in enzyme activity. Most enzymes that display allosteric regulation are metabolic enzymes involved in the degradation or...