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Updated: Jun 26, 2025

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Biocatalysis with In-Situ Product Removal Improves p-Coumaric Acid Production.

Alexander Virklund1, Alex Toftgaard Nielsen2, John M Woodley1

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|May 14, 2024
PubMed
Summary

Biocatalysis for p-coumaric acid production was enhanced using in situ product removal. This method, employing an aqueous two-phase system, improved yields by 1.9-fold, overcoming enzyme inhibition challenges.

Keywords:
BiocatalysisBiphasic catalysisIn-situ product removalammonia lyaseimmobilization

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Area of Science:

  • Biotechnology
  • Biochemical Engineering
  • Enzyme Catalysis

Background:

  • Natural p-coumaric acid has valuable applications but bioprocessing yields are insufficient.
  • Biocatalysis offers promise but is limited by product inhibition of tyrosine ammonia lyase.
  • Decoupling growth and production is a key strategy for improving biocatalytic efficiency.

Purpose of the Study:

  • To revisit and improve a biocatalytic process for p-coumaric acid production.
  • To address product inhibition of tyrosine ammonia lyase (TAL).
  • To implement in situ product removal (ISPR) using an aqueous two-phase system (ATPS).

Main Methods:

  • A polymer/salt aqueous two-phase system was identified for p-coumaric acid extraction.
  • The ATPS was optimized for TAL activity, considering salt concentration and polymer type.
  • An immobilized TAL biocatalyst was used within the selected ATPS for ISPR.

Main Results:

  • An ATPS using 5% potassium phosphate and 12.5% poly(ethylene glycol-ran-propylene glycol) was selected.
  • The optimized ATPS allowed for efficient extraction of p-coumaric acid with a partition coefficient up to 13.
  • The immobilized TAL biocatalyst in ATPS achieved 33.5 g/L p-coumaric acid in 24 hours, a 1.9-fold increase.

Conclusions:

  • In situ product removal using an optimized aqueous two-phase system effectively overcomes TAL product inhibition.
  • This enhanced biocatalytic process significantly improves p-coumaric acid production metrics.
  • The developed method presents a viable strategy for efficient and scalable production of p-coumaric acid.