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Advances in in vivo continuous evolution technologies for strain development and enzyme engineering.

Yuan Liu1, Cong Gao1, Xiaomin Li1

  • 1School of Biotechnology and Key Laboratory of Industrial Biotechnology of the Ministry of Education, Jiangnan University, Wuxi, China; Key Laboratory of Industrial Synthetic Biology of Jiangsu Province, Jiangnan University, Wuxi, China.

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In vivo continuous evolution accelerates microbial strain and enzyme engineering by increasing mutation rates within cells. This review explores key platforms, offering guidance for enhanced industrial applications.

Keywords:
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Area of Science:

  • Microbiology
  • Biotechnology
  • Synthetic Biology

Background:

  • Evolutionary engineering is crucial for developing industrial microbial strains with improved stress tolerance and productivity.
  • Current limitations include low natural mutation rates, small mutation libraries, and slow screening processes.

Purpose of the Study:

  • To systematically review in vivo continuous evolution platforms for microbial engineering.
  • To compare different mutational mechanisms, their design principles, applications, and optimization strategies.

Main Methods:

  • Examination of three in vivo continuous evolution platforms: error-prone DNA replication, DNA base modification, and DNA recombination.
  • Analysis of underlying molecular mechanisms, advantages, and limitations of each platform.

Main Results:

  • In vivo continuous evolution significantly elevates mutation rates, enabling rapid strain and gene evolution.
  • Discussion of strategies for optimizing mutational spectra and selection regimes for specific applications.

Conclusions:

  • In vivo continuous evolution offers a powerful approach to overcome limitations in traditional evolutionary engineering.
  • This review provides a framework for understanding these technologies and guiding future strain development and enzyme engineering efforts.