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Multidimensionally engineered Escherichia coli for efficient gadusol biosynthesis with high-throughput quantitative

Ping Zhang1, Weijiao Zhang1, Haibo Xiong1

  • 1Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China; Science Center for Future Foods, Jiangnan University, Wuxi 214122, China; Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.

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Summary

Researchers engineered *Escherichia coli* for efficient biosynthesis of the sunscreen compound gadusol. This microbial production strategy achieved a final titer of 4.2 g/l, overcoming previous availability limitations.

Keywords:
antioxidantcofactor engineeringgadusolhigh-throughput quantitative analysissunscreen

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Area of Science:

  • Biotechnology
  • Synthetic Biology
  • Metabolic Engineering

Background:

  • Gadusol, a cyclohexenone, possesses valuable sunscreen and antioxidant properties.
  • Limited availability of gadusol, particularly from microbial sources, restricts its widespread application.
  • Microbial cell factories offer a promising avenue for sustainable gadusol production.

Purpose of the Study:

  • To engineer *Escherichia coli* for efficient biosynthesis of gadusol.
  • To develop high-throughput methods for gadusol quantification and strain screening.
  • To establish a robust microbial production platform for gadusol.

Main Methods:

  • Reprogramming of the gadusol biosynthesis pathway in *E. coli*.
  • Development of a colorimetric assay for high-throughput screening.
  • Small RNA library screening to identify regulatory targets.
  • Directed evolution of transketolase A and cofactor optimization.
  • Fed-batch fermentation for large-scale production.

Main Results:

  • Initial gadusol titer of 45.2 mg/l achieved after pathway reprogramming.
  • Titer increased to 700 mg/l through small RNA screening and regulatory target identification.
  • Further optimization led to a titer of 965 mg/l via enzyme evolution and cofactor balancing.
  • Final production reached 4.2 g/l using a fed-batch fermentation strategy.

Conclusions:

  • Systematic engineering of *E. coli* enabled efficient gadusol biosynthesis.
  • The developed framework facilitates rapid screening and construction of microbial cell factories.
  • This study provides a scalable method for producing gadusol, addressing its availability challenges.