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Glycosyltransferase engineering for carbohydrate synthesis.

John B McArthur1, Xi Chen2

  • 1Department of Chemistry, University of California, Davis, CA 95616, U.S.A.

Biochemical Society Transactions
|February 11, 2016
PubMed
Summary

Enzyme engineering of glycosyltransferases (GTs) enhances their utility for synthesizing complex carbohydrates. Structure-guided directed evolution (SGDE) optimizes GTs for biotechnological applications, improving carbohydrate production.

Keywords:
carbohydratedirected evolutionenzymatic synthesisglycosyltransferasemutagenesisprotein engineering

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Area of Science:

  • Biochemistry
  • Enzymology
  • Carbohydrate Chemistry

Background:

  • Glycosyltransferases (GTs) are crucial enzymes for synthesizing complex carbohydrates with significant biological roles.
  • Wild-type GTs often require modifications for efficient large-scale production of natural and non-natural carbohydrates.
  • Advances in GT structural biology provide opportunities for enzyme engineering.

Purpose of the Study:

  • To review progress in engineering glycosyltransferases (GTs) for improved functionality.
  • To highlight the success of structure-guided rational design and directed evolution in GT engineering.
  • To discuss future directions for GT engineering and assay development.

Main Methods:

  • Utilizing crystal structures of GTs with substrate analogues for rational design.
  • Employing directed evolution to generate diverse GT mutants.
  • Implementing structure-guided directed evolution (SGDE) by targeting substrate-binding sites.
  • High-throughput screening of engineered GT variants.

Main Results:

  • Crystal structure-guided rational design has successfully yielded GT mutants with enhanced properties.
  • Directed evolution provides a complementary approach for discovering novel GT functionalities.
  • SGDE effectively explores the biotechnological potential of GTs through targeted mutagenesis.
  • Significant progress has been made in engineering GTs for diverse applications.

Conclusions:

  • Enzyme engineering, particularly SGDE, is vital for unlocking the full biotechnological potential of glycosyltransferases.
  • Combining rational design with directed evolution accelerates the development of tailored GTs.
  • Future efforts should focus on improving structure-activity relationship understanding and assay development for GT engineering.