研究双点表面附着对酶稳定性和活性的影响
Journal of the American Chemical Society
|November 8, 2018
概括
在固体支上固定时,具有两个表面结合点的工程酶变体表现出增强的稳定性. 这种合理的设计方法提高了生物技术应用中的酶性能.
科学领域:
- 生物催化和酶工程
- 表面化学和纳米技术
- 计算生物学和分子建模
背景情况:
- 在固体支上的酶固定增强了稳定性,并简化了生物技术应用的处理.
- 表面固定可以对酶结构产生负面影响,导致催化效率和稳定性降低.
- 需要合理的设计来优化酶固定策略.
研究的目的:
- 在表面固定后改进稳定性的酶变体.
- 展示一种结合模拟和实验方法的系统方法,以优化酶固定.
- 研究战略位置的结合点对酶稳定性和定向的影响.
主要方法:
- 分子动力学 (MD) 模拟设计表面绑定站点.
- 具有特定基基残留的基还原酶 (NfsB) 变体的工程.
- 在maleimide终端的自组装单层 (SAM) 表面上进行共价固定.
- 总频率生成 (SFG) 和减弱的全反射里埃变换红外光谱 (ATR-FTIR) 来确定酶方向.
- 热稳定性分析以量化改进.
主要成果:
- MD模拟成功指导了有效的绑定站点的设计.
- 设计的NfsB变种具有两个绑定点,在固定时显示出更好的热稳定性.
- 不动化的NfsB变体呈现出与模拟预测一致的方向.
- 与单位固定相比,通过两个位点固定酶的稳定性至少增加了60%.
结论:
- 经过战略设计的表面结合点在固定过程中提高了酶的稳定性.
- 一种结合模拟和实验方法为优化酶固定提供了合理的方法.
- 这种策略在各种应用中广泛适用于提高表面固定酶的性能.
相关概念视频
Enzymes and Activation Energy
23.6K
The activation energy (or free energy of activation), abbreviated as Ea, is the small amount of energy input necessary for all chemical reactions to occur. During chemical reactions, certain chemical bonds break, and new ones form. For example, when a glucose molecule breaks down, bonds between the molecule's carbon atoms break. Since these are energy-storing bonds, they release energy when broken. However, the molecule must be somewhat contorted to get into a state that allows the bonds to...
23.6K
Enzymes
94.7K
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...
Enzyme deficiencies can often translate into life-threatening diseases. For example, a genetic abnormality resulting in the deficiency of the enzyme G6PD...
94.7K
Attachment
522
Attachment is vital for infant development, as warm social interactions support growth and well-being. In a classic 1958 study by Harry Harlow, the significance of warmth and comfort in forming attachments was examined. Harlow separated newborn monkeys from their mothers and provided two artificial "mothers": one made of cold wire and the other covered in soft cloth. Despite the wire mother offering food, the infant monkeys preferred the comfort of the cloth mother, demonstrating that...
522
RNA Stability
35.7K
Intact DNA strands can be found in fossils, while scientists sometimes struggle to keep RNA intact under laboratory conditions. The structural variations between RNA and DNA underlie the differences in their stability and longevity. Because DNA is double-stranded, it is inherently more stable. The single-stranded structure of RNA is less stable but also more flexible and can form weak internal bonds. Additionally, most RNAs in the cell are relatively short, while DNA can be up to 250 million...
35.7K
Enzyme-linked Receptors
86.6K
Enzyme-linked receptors are proteins that act as both receptor and enzyme, activating multiple intracellular signals. This is a large group of receptors that include the receptor tyrosine kinase (RTK) family. Many growth factors and hormones bind to and activate the RTKs.
Neurotrophin (NT) receptors are a family of RTKs, including trkA, trkB, and trkC (tropomyosin-related kinase) receptors. TrkA is specific for nerve growth factor (NGF), neurotrophin-6, and neurotrophin-7. TrkB binds...
Neurotrophin (NT) receptors are a family of RTKs, including trkA, trkB, and trkC (tropomyosin-related kinase) receptors. TrkA is specific for nerve growth factor (NGF), neurotrophin-6, and neurotrophin-7. TrkB binds...
86.6K
Enzyme Kinetics
104.1K
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...
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...
104.1K


