相关实验视频

Updated: Jun 28, 2025

Concanavalin A-Based Sedimentation Assay to Measure Substrate Binding of Glucan Phosphatases

Concanavalin A-Based Sedimentation Assay to Measure Substrate Binding of Glucan Phosphatases

Published on: December 23, 2022

粉加工α-氨酶的结构和功能洞察力来自超热友的archaeon Pyrococcusabyssisi

Mohsin Shad¹, Hafiz Muzzammel Rehman², Muhammad Waheed Akhtar³

¹School of Biological Sciences, University of the Punjab, Quaid-e-Azam Campus, P.O. 54590, Lahore, Pakistan; Structural Biology, The Rosalind Franklin Institute, Harwell Science & Innovation Campus, Didcot, OX11 0QS, United Kingdom.

Carbohydrate research

|April 24, 2024

概括

一种来自Pyrococcus abyssi的新型超热友性α-氨酶 (PaAFG) 已被识别和特征. 这种热稳定酶在90°C和pH 6处表现出最佳活性,显示出对工业应用的前景.

科学领域:

生物化学生物化学
酶学是一种酶学.
结构生物学结构生物学

背景情况:

对超热友性Pyrococcus abyssi的基因组查揭示了新的α-amylase序列.
已识别的α-amylase (PaAFG) 具有一个N端的GH57催化域,一个中心域和一个C端域.

研究的目的:

进行新型alpha-amylase的in-silico结构和功能分析.
为了实现酶的重组表达和特征.
为了研究其结构性质和带结合能力.

主要方法:

用于结构分析的同质模型.
在大肠杆菌罗塞塔TM (DE3) pLysS中的重组表达.
描述最佳温度,pH值和运动参数 (Km,Vmax).
循环二重化 (CD) 光谱用于结构稳定性.
分子动力学模拟用于稳定性和移动性的评估.

主要成果:

PaAFG成功地以可溶性形式表达和净化.
在90°C和pH 6.0下观察到最佳活性.
动力分析显示,小麦粉的Km为0.20 ± 0.053毫克,Vmax为25.00 ± 0.67μmol-1毫克-1.1.
CD光谱和分子动力学模拟证实了酶的热稳定性和结构完整性.

结论:

来自P. abyssi的新型α-氨酶PaAFG是一种具有广泛基质特异性的热稳定酶.
它的可溶性生产,高最佳温度和稳定性使其成为各种工业应用的强有力的候选者.

关键词:

电磁盘光谱学CD光谱学超热爱的人是超热爱的人.模拟MD的模拟方法分子对接是分子对接.这是一种Pyrococcus abyssi.α-氨基酶的使用.

更多相关视频

Transverse Sectioning of Mature Rice Oryza sativa L. Kernels for Scanning Electron Microscopy Imaging Using Pipette Tips as Immobilization Support

Transverse Sectioning of Mature Rice Oryza sativa L. Kernels for Scanning Electron Microscopy Imaging Using Pipette Tips as Immobilization Support

Published on: January 25, 2022

Determination of Glucan Chain Length Distribution of Glycogen Using the Fluorophore-Assisted Carbohydrate Electrophoresis FACE Method

Determination of Glucan Chain Length Distribution of Glycogen Using the Fluorophore-Assisted Carbohydrate Electrophoresis FACE Method

Published on: March 31, 2022

相关概念视频

Carbohydrate Digestion

Carbohydrate Digestion

Carbohydrate digestion and metabolism break down simple and complex carbohydrates from food into saccharides (i.e., sugars) for the body to use as energy. Carbohydrate digestion starts in the mouth during mastication, or chewing. The masticated carbohydrates remain intact in the stomach. Digestion resumes in the duodenum of the small intestine, where pancreatic alpha-amylase and brush border enzymes of the microvilli convert complex carbohydrates to monosaccharides. Finally, the monosaccharides...

Allosteric Proteins-ATCase

Allosteric Proteins-ATCase

Binding sites linkages can regulate a protein's function. For example, enzyme activity is often regulated through a feedback mechanism where the end product of the biochemical process serves as an inhibitor.
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis...

Introduction to Carbohydrates

Introduction to Carbohydrates

Carbohydrates, proteins, and fats are the primary macronutrients in the human diet. However, carbohydrates are the most favored source of energy in the body. They can be found in a wide variety of foods, including whole grains, fruit, and vegetables, in various forms, such as sugars, starch, and dietary fiber. Based on their structure, carbohydrates are classified into three main classes— monosaccharides, disaccharides, and polysaccharides. The body's cells can only utilize simple...