相关实验视频

Updated: Aug 6, 2026

Engineering Cell-permeable Protein

Engineering Cell-permeable Protein

Published on: December 28, 2009

由IPTG诱导的高蛋白表达,用于L-phosphinothricin的全细胞生物合成

Jian-Miao Xu^1,2,3, Zhou-Sheng Wu^1,2,3, Ke-Ji Zhao^1,2,3

¹Institute of Bioengineering, Zhejiang University of Technology, Hangzhou, P. R. China.

Biotechnology journal

|June 2, 2023

概括

这项研究优化了使用工程Escherichia coli菌株的L-phosphinothricin (L-PPT) 生物催化生产. 在酶活性和生物质方面,IPTG诱导被证明优于乳糖,使有效的L-PPT合成成为可能.

科学领域:

生物技术是生物技术.
酶工程是什么? 酶工程是什么?
工业微生物学工业微生物学

背景情况:

通过生物催化剂生产L-phosphinothricin (L-PPT) 是一种有希望的可持续方法.
这项研究使用了两步的酶过程,涉及工程Escherichia coli菌株.

研究的目的:

为了优化L-PPT的生物催化生产.
为了比较IPTG和乳糖作为诱导酶表达和活性在工程化大肠杆菌菌株中的疗效.

主要方法:

在大肠杆菌中,D-氨基酸氧化酶和催化酶 (DAAO-CAT) 的同时表达,用于D-PPT氧化.
在大肠杆菌中,谷氨酸脱酶和形式脱酶 (GluDH-FDH) 的同时表达,用于将PPO降低到L-PPT.
在5L发酵器中优化诱导条件 (IPTG度,时间,温度).

主要成果:

大肠杆菌DAAO-CAT的最佳条件:0.05mM IPTG,28°C的18小时,DAAO的特异活性为153.20U/g,CAT的特异活性为896.23U/g.
大肠杆菌GluDH-FDH的最佳条件:0.2 mM IPTG,28°C的19小时,产生GluDH的41.72 U/g和FDH的109.70 U/g的特异活性.
实现了高转换率 (>99%):D-PPT转为PPO的9.0 g·L−1·h−1和PPO转为L-PPT的14.5 g·L−1·h−1.

结论:

在两种大肠杆菌菌株中,IPTG在酶活性和生物质方面比乳糖具有优势.
IPTG是一种更环保和经济可行的诱导剂,用于扩大工业L-PPT发酵.
开发的生物催化工艺是迄今为止L-PPT生产中最有效的方法.

关键词:

D-氨基酸氧化酶的作用在IPTG中使用IPTG.这是一种L-phosphinothricin.发酵发酵发酵发酵谷氨酸脱酶的使用

更多相关视频

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

Targeting Cysteine Thiols for in Vitro Site-specific Glycosylation of Recombinant Proteins

Targeting Cysteine Thiols for in Vitro Site-specific Glycosylation of Recombinant Proteins

Published on: October 4, 2017

相关概念视频

Stringent Response in E. coli

Stringent Response in E. coli

Bacterial growth is closely tied to nutrient availability, with cells proliferating exponentially under favorable conditions and entering a stationary phase when resources become scarce. This transition is mediated by a regulatory mechanism known as the stringent response, which allows bacteria to adapt to nutrient deprivation by modulating gene expression and metabolic activity.During nutrient scarcity, intracellular amino acid levels decline. It results in the accumulation of uncharged tRNAs...

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...