School of Chemical Engineering & Technology, Tianjin University, Tianjin;ey Laboratory of Systems Bioengineering Ministry of Education, Tianjin;Collaborative Innovation Center of Chemical Science and Engineering(Tianjin), Tianjin,School of Chemical Engineering & Technology, Tianjin University, Tianjin;ey Laboratory of Systems Bioengineering Ministry of Education, Tianjin;Collaborative Innovation Center of Chemical Science and Engineering(Tianjin), Tianjin,School of Chemical Engineering & Technology, Tianjin University, Tianjin;ey Laboratory of Systems Bioengineering Ministry of Education, Tianjin;Collaborative Innovation Center of Chemical Science and Engineering(Tianjin), Tianjin,School of Chemical Engineering & Technology, Tianjin University, Tianjin;ey Laboratory of Systems Bioengineering Ministry of Education, Tianjin;Collaborative Innovation Center of Chemical Science and Engineering(Tianjin), Tianjin,School of Chemical Engineering & Technology, Tianjin University, Tianjin;ey Laboratory of Systems Bioengineering Ministry of Education, Tianjin;Collaborative Innovation Center of Chemical Science and Engineering(Tianjin), Tianjin
This work was supported by grants from The National Basic Research Program of China (2011CBA00804, 2012CB725203), The National Natural Science Foundation of China (21206112, 21390201), Hi-Tech Research and Development Program of China (2012AA022103, 2012AA02A702) and The Innovation Foundation of Tianjin University (1308)
Protein directed evolution is widely used to improve enzymes, particularly for industrial biocatalytic processes and construction of cell factories. It is an efficient and powerful tool to improve and optimize natural proteins in order to generate robust biocatalysts for practical applications. In addition, optimization of metabolic pathways, regulation of functional regulatory systems, and development of desired complex phenotypes in industrial host organisms have all been achieved by way of protein directed evolution. Numerous in vivo and in vitro methods have been developed for the efficient evolutionary effects, especially in high mutation rate and rapidly high-throughput screening capabilities. Some of the methods have only recently been applied for general use and are just beginning to find greater application. In this review, we summarize some of the new methods for mutant libraries generation, including random evolution, semi-rational evolution and rational evolution. And current state-of-the-art screening techniques in protein directed evolution are also reviewed. Advancements are discussed with respect to the state of the art in diversity generation and high-throughput screening capabilities. Meanwhile, limitations and remaining challenges are also pronounced.
WANG Xiao-Yue, WANG Bai-Yun, WANG Zhi-Wen, CHEN Tao, ZHAO Xue-Ming. The Research Progress of Protein Directed Evolution[J]. Progress in Biochemistry and Biophysics,2015,42(2):123-131
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