相关实验视频

Updated: May 12, 2026

Expression, Isolation, and Purification of Soluble and Insoluble Biotinylated Proteins for Nerve Tissue Regeneration

Expression, Isolation, and Purification of Soluble and Insoluble Biotinylated Proteins for Nerve Tissue Regeneration

Published on: January 22, 2014

蛋白质工程,表达的优化,以及从Alkalihalobacillus clausii FYX FYX中性蛋白酶的应用

Yuxi Fu¹, Yi Rao¹, Yongqing Liao¹

¹State Key Laboratory of Biocatalysis and Enzyme Engineering, Environmental Microbial Technology Center of Hubei Province, College of Life Sciences, Hubei University, Wuhan 430062, PR China.

International journal of biological macromolecules

|March 10, 2025

概括

研究人员设计了一种性蛋白酶 (AE0) 以提高性能. 突变体显示出增强的热稳定性和催化效率,其中一个三重突变体在工业应用中实现了178.5%的活性增加.

科学领域:

生物技术和工业微生物学
酶工程和蛋白质科学酶工程和蛋白质科学

背景情况:

蛋白酶是重要的工业酶,用于洗剂,食品和农业.
提高性蛋白酶的性能是扩大其市场应用的关键.
新型性蛋白酶的表征对于进一步发展至关重要.

研究的目的:

来自Alkalihalobacillus clausii FYX的性蛋白酶AE0的特征.
通过半理性设计和推动器工程来提高AE0的热稳定性和催化效率.
为工业应用开发一个高效的表达系统.

主要方法:

性蛋白酶AE0在Bacillus licheniformisDW2△aprE中进行了表征.
半理性设计被用来创造六个单个突变.
引入了进一步的突变,产生了双重和三重突变 (AE0DM1,AE0MM2).
促进器工程被用来增强酶活性.
评估了热稳定性和运动参数.

主要成果:

野生型AE0在60°C和pH值10.5.5时表现出最佳活动.
单个突变G113I表现出最高的特异活性 (28,150.64 U/mg).
三重突变AE0MM2的特定活性达到31,868.56U/mg.
突变G113I,H118D和N167S增强了热稳定性和催化效率.
推动者工程使AE0MM2活动增加了178.5% (36,685.33U/mL).
AE0MM2证明了有效的豆粉水解.

结论:

工程性蛋白酶AE0表现出更好的热稳定性和催化效率.
开发的表达系统显著提高了工业用途的酶生产.
这项研究为各种工业应用提供了强大的性蛋白酶.

关键词:

阿尔卡利哈洛巴西勒斯克劳西 (Alkalihalobacillus clausii) FYXX FYX FYX FYX FYX FYX FYX FYX 性蛋白酶是一种性蛋白酶.表达方式优化表达方式优化一个半理性的设计.大豆粉蛋白质降解

更多相关视频

Generic Protocol for Optimization of Heterologous Protein Production Using Automated Microbioreactor Technology

Generic Protocol for Optimization of Heterologous Protein Production Using Automated Microbioreactor Technology

Published on: December 15, 2017

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

相关概念视频

Upstream Processing

Upstream Processing

Upstream processing represents a critical phase in biomanufacturing, wherein biological systems such as microorganisms, mammalian cells, or insect cells are cultivated to produce therapeutic proteins, vaccines, enzymes, or other biologically derived products. This phase encompasses all steps from the selection and genetic manipulation of the production organism to the cultivation of cells in bioreactors under tightly controlled environmental conditions.Host Selection and Genetic OptimizationThe...

Production of Pharmaceuticals

Production of Pharmaceuticals

Industrial insulin production uses genetically engineered E. coli expressing a proinsulin gene controlled by a tryptophan promoter and containing a methionine linker for later cleavage. The cells also carry ampicillin resistance for selective growth. Seed cultures are stored at −80 °C and production begins by thawing a small amount to inoculate starter cultures, which are progressively scaled to a 50,000-L bioreactor. In the bioreactor, E. coli grow in nutrient-rich media under sterile, tightly...