相关实验视频

Updated: Mar 17, 2026

Engineering 'Golden' Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence

Engineering 'Golden' Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence

Published on: April 27, 2018

通过in vivo定向进化对高水平的l-氨酸生产进行工程氨酸合成

Xingyu Zhu^1,2, Hengwei Zhang^1,3, Di Zhang¹

¹Key Laboratory of Industrial Biotechnology of the Ministry of Education, School of Biotechnology, Laboratory of Applied Microorganisms and Metabolic Engineering, Jiangnan University, Wuxi 214122, China.

Journal of agricultural and food chemistry

|March 15, 2026

概括

研究人员使用酸合成酶 (TrpS) 改进了l-氨酸的工业生产. 定向进化产生了优越的TrpS突变,实现了高的l-氨酸位和高效的全细胞固定,以增强生物合成.

科学领域:

生物催化剂是一种生物催化剂.
酵素工程是什么? 酶工程是什么
代谢工程是代谢工程.

背景情况:

从l-serine和硫化物直接合成l-cysteine是由酸盐合成酶 (TrpS) 催化.
目前的TrpS效率在高基质度下是有限的,阻碍了工业应用.
提高TrpS的催化效率对于具有成本效益的l-氨酸生产至关重要.

研究的目的:

开发一种定向进化平台,用于改善l-氨酸生物合成的氨酸合成酶 (TrpS).
为了识别具有增强催化活性和稳定性的TrpS变体.
建立一个工业上可行的生产l-氨酸的工艺.

主要方法:

建立了一个定向进化平台,结合了体内连续的突变发生.
一个高度敏感的l-氨酸生物传感器和光激活细胞分类 (FACS) 用于高通量选.
采用全细胞固定化策略,以提高生物催化剂的稳定性和可重复使用性.

主要成果:

成功生成多个TrpS变体,V139M/A302P突变体显著提高了活性和稳定性.
在优化条件下,工程TrpS突变物达到116.4g/L的l-氨酸标位.
固定细胞产生最大的l-氨酸位为113.05g/L,单一批的转化率为93.3%.

结论:

开发的定向进化平台对选改进的生物催化剂非常有效.
工程TrpS突变和固定化策略为工业l-氨酸生物合成提供了一个有前途的途径.
这项研究提供了一种强大且可扩展的生产l-氨酸的方法.

关键词:

指导进化是指导进化的.固定化固定化固定化一种类型的L-氨酸.传感器传感器传感器托芬合成酶的使用方法

更多相关视频

PCR Mutagenesis, Cloning, Expression, Fast Protein Purification Protocols and Crystallization of the Wild Type and Mutant Forms of Tryptophan Synthase

PCR Mutagenesis, Cloning, Expression, Fast Protein Purification Protocols and Crystallization of the Wild Type and Mutant Forms of Tryptophan Synthase

Published on: September 26, 2020

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

Amino Acid Biosynthetic Pathways

Amino Acid Biosynthetic Pathways

Amino acid biosynthesis is essential for cell growth, protein synthesis, and metabolic regulation. Cells generate essential and non-essential amino acids from metabolic intermediates to sustain vital biological functions. These intermediates originate from key metabolic pathways: glycolysis, the tricarboxylic acid (TCA) cycle, and the pentose phosphate pathway. Important precursors include α-ketoglutarate, pyruvate, oxaloacetate, phosphoenolpyruvate, and erythrose-4-phosphate, which...