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[Enzyme ancestral sequence reconstruction and directed evolution].

Kun Zhang1, Yifei Dai1, Jindi Sun1

  • 1College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, Jiangsu, China.

Sheng Wu Gong Cheng Xue Bao = Chinese Journal of Biotechnology
|January 5, 2022
PubMed
Summary

Ancestral sequence reconstruction (ASR) computationally deduces extinct enzyme sequences. These reconstructed ancestral enzymes often exhibit enhanced stability, making them valuable for protein engineering and directed evolution studies.

Keywords:
ancestral enzymeancestral sequence reconstructionbiocatalysisdirected evolutionenzyme structure-function relationships

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Area of Science:

  • Biochemistry
  • Evolutionary Biology
  • Computational Biology

Background:

  • Enzymes are crucial biocatalysts involved in numerous biological processes.
  • Understanding enzyme evolution and adaptation is key to deciphering molecular mechanisms.
  • Extinct organisms' enzyme sequences can be inferred using computational methods.

Purpose of the Study:

  • To summarize the methodology, applications, and software for ancestral sequence reconstruction (ASR).
  • To highlight the utility of ASR in studying enzyme evolution and adaptation.
  • To discuss the potential of ancestral enzymes in directed evolution.

Main Methods:

  • Ancestral Sequence Reconstruction (ASR) involves six key steps.
  • These steps include sequence collection, multiple sequence alignment, phylogenetic tree construction, computational sequence deduction, gene cloning, and property characterization.
  • In silico approaches are central to deducing ancestral enzyme sequences.

Main Results:

  • ASR enables the study of molecular adaptation to environmental changes over geological timescales.
  • Reconstructed ancestral enzymes frequently display superior temperature and mutation stability.
  • This enhanced stability makes ancestral enzymes promising scaffolds for protein engineering.

Conclusions:

  • ASR is a powerful tool for investigating enzyme sequence-structure-function relationships.
  • Ancestral enzymes offer improved stability, ideal for directed evolution applications.
  • This approach has significant potential for enzyme engineering and biotechnological advancements.