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Development and optimization of an engineered E. coli platform for nitrotryptophan biosynthesis.

Haiying Lin1, Man Zhang2, Wenbin Kang2

  • 1College of Biological Science and Engineering, Fuzhou University, Fuzhou 350108, China; The Key Laboratory of Novel Enzyme Design and Creation of Fujian Province, Fuzhou 350116, China.

Enzyme and Microbial Technology
|November 23, 2025
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Summary

This study developed a novel microbial bioreactor for direct nitrotryptophan synthesis. The engineered Escherichia coli efficiently produces nitrotryptophan isomers using a P450 enzyme and nitric oxide synthetase.

Keywords:
BiosynthesisMetabolic pathwayNitrotryptophanP450 enzymeSodium nitroprusside

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Area of Science:

  • Biocatalysis and metabolic engineering
  • Synthetic biology
  • Chemical synthesis

Background:

  • Nitrotryptophan and its derivatives are key intermediates in synthesizing bioactive compounds and functional materials.
  • Direct aromatic nitration of tryptophan is challenging using traditional chemical methods.

Purpose of the Study:

  • To engineer an efficient biocatalytic bioreactor in Escherichia coli for direct synthesis of nitrotryptophan isomers.
  • To establish a microbial platform for producing valuable nitroaromatic compounds.

Main Methods:

  • Engineered Escherichia coli with a P450 enzyme (TB14) and nitric oxide synthetase (BsNOS) or sodium nitroprusside (SNP) for direct nitration.
  • Incorporated glucose dehydrogenase (GDH) for NADPH regeneration.
  • Optimized pathway balancing, fermentation conditions, and eliminated competing metabolic pathways.

Main Results:

  • Achieved an initial nitrotryptophan titer of 133.2 mg/L.
  • Increased the final titer to 209.9 mg/L within 48 hours through systematic optimization.
  • Demonstrated efficient microbial production of nitrotryptophan isomers.

Conclusions:

  • Developed a robust biocatalytic platform for direct microbial nitration of L-tryptophan.
  • Provided insights for future biocatalytic nitration strategies of aromatic compounds.
  • Successfully synthesized nitrotryptophan isomers using an engineered Escherichia coli strain.