相关实验视频

Updated: Jul 2, 2025

Functional Complementation Analysis FCA: A Laboratory Exercise Designed and Implemented to Supplement the Teaching of Biochemical Pathways

Functional Complementation Analysis FCA: A Laboratory Exercise Designed and Implemented to Supplement the Teaching of Biochemical Pathways

Published on: June 24, 2016

来自Geobacillus thermopakistaniensis的一个L-asparaginase的结构和功能分析

Ayesha Sania¹, Majida Atta Muhammad¹, Muhammad Sajed¹

¹School of Biological Sciences, University of the Punjab, Quaid-e-Azam Campus, Lahore 54590, Pakistan.

International journal of biological macromolecules

|February 26, 2024

概括

鉴定了Geobacillus thermopakistaniensis L-asparaginase (ASNaseGt) 的特征,并发现它是一种高度恒温的细胞质I型酶,而不是预测的II型酶. 它的四重体形式在52°C和pH值9.5.5时表现出最佳活性.

科学领域:

酶学是一种酶学.
结构生物学结构生物学
微生物生物化学微生物生物化学

背景情况:

在Geobacillus thermopakistaniensis的基因组中编码了一个假定的II型L-asparaginase (ASNaseGt).
最初的结构分析表明ASNaseGt可能作为I型L-asparaginase而起作用.

研究的目的:

确定ASNaseGt的确切分类为I型或II型.
描述重组ASNaseGt.的结构和功能性质.
为了研究ASNaseGt在G. thermopakistaniensis中的细胞局部化.

主要方法:

再组合蛋白的表达和净化.
酶活性在不同温度和pH值下进行测定.
动力参数 (Vmax,Km) 的确定.
热稳定性测试. 热稳定性测试.
使用β-mercaptoethanol进行寡合化状态分析.
通过G. thermopakistaniensis进行细胞局部化研究.

主要成果:

ASNaseGt在52°C和pH值9.5时表现出最佳活性,在β-mercaptoethanol的存在下活性增加.
该酶表现出高温稳定性,在55°C下6小时后保持超过80%的活性.
亚酶Gt的四重体形式显示出最高的特异活性 (4300 U/mg).
局部化研究证实ASNaseGt是一种细胞质蛋白.

结论:

结构和功能特征,以及局部化数据,最终确定ASNaseGt为I型L-asparaginase.
ASNaseGt是一种热稳定的细胞酶,在生理条件下具有显著的活性.

关键词:

G. thermopakistaniensisensis 这种植物是印度的.在L-asparaginase的使用中.寡合化是指一种氧化物.结构分析结构分析热稳定性热稳定性

更多相关视频

The Use of a β-lactamase-based Conductimetric Biosensor Assay to Detect Biomolecular Interactions

The Use of a β-lactamase-based Conductimetric Biosensor Assay to Detect Biomolecular Interactions

Published on: February 1, 2018

Homogeneous Glycoconjugate Produced by Combined Unnatural Amino Acid Incorporation and Click-Chemistry for Vaccine Purposes

Homogeneous Glycoconjugate Produced by Combined Unnatural Amino Acid Incorporation and Click-Chemistry for Vaccine Purposes

Published on: December 19, 2020

相关概念视频

Ligand Binding and Linkage

Ligand Binding and Linkage

Allosteric proteins have more than one ligand binding site; the binding of a ligand to any of these sites influences the binding of ligands to the other sites. When a protein is allosteric, its binding sites are called coupled or linked. In the case of enzymes, the site that binds to the substrate is known as the active site and the other site is known as the regulatory site. When a ligand binds to the regulatory site, this leads to conformational changes in the protein that can influence...