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Precursor-directed biosynthesis of 5-hydroxytryptophan using metabolically engineered E. coli.

Xinxiao Sun1, Yuheng Lin2, Qipeng Yuan1

  • 1†State Key Laboratory of Chemical Resource Engineering, Beijing University of Chemical Technology, Beijing 100029, China.

ACS Synthetic Biology
|October 3, 2014
PubMed
Summary
This summary is machine-generated.

Researchers developed a novel biosynthetic pathway for producing 5-hydroxytryptophan (5-HTP) from glucose. This method utilizes enzyme promiscuity to create 5-HTP efficiently without unstable enzymes.

Keywords:
5-hydroxyanthranilate5-hydroxytryptophanprecursor-directed biosynthesissalicylate 5-hydroxylase

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Area of Science:

  • Synthetic Biology
  • Metabolic Engineering
  • Enzyme Engineering

Background:

  • 5-hydroxytryptophan (5-HTP) is a crucial precursor for serotonin and melatonin.
  • Existing methods for 5-HTP production face challenges due to enzyme instability and pathway complexity.

Purpose of the Study:

  • To design and validate a novel biosynthetic pathway for de novo 5-HTP production from glucose.
  • To leverage enzyme promiscuity for creating efficient and stable metabolic pathways.

Main Methods:

  • Reverse pathway design starting from 5-HTP.
  • Utilized E. coli TrpDCBA for 5-HTP production from 5-hydroxyanthranilate (5-HI).
  • Employed a novel salicylate 5-hydroxylase for converting anthranilate to 5-HI.
  • Modular optimization for 5-HI production from glucose.
  • Two-stage strategy for de novo 5-HTP synthesis.

Main Results:

  • Achieved high-titer production of 5-HTP from 5-HI.
  • Successfully converted anthranilate to 5-HI using a novel enzyme.
  • Optimized and demonstrated 5-HI production from glucose.
  • Established a complete de novo 5-HTP production pathway from glucose.

Conclusions:

  • Demonstrated the successful application of enzyme promiscuity in designing non-natural biosynthetic pathways.
  • Established a novel and efficient microbial cell factory for 5-HTP production.
  • This pathway offers a promising alternative for sustainable 5-HTP manufacturing.