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Jesús E Rodríguez Gamero1, Lorenzo Favaro1, Valentino Pizzocchero1

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Engineered bacteria expressing a Staphylococcus aureus nuclease gene significantly reduce the viscosity of polyhydroxyalkanoates (PHAs) lysates. This innovation offers a cost-effective solution for PHA downstream processing in industrial manufacturing.

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Area of Science:

  • Biotechnology
  • Microbial Engineering
  • Biopolymer Production

Background:

  • Industrial polyhydroxyalkanoates (PHAs) production involves purifying PHAs granules from high-cell-density cultures.
  • Cell lysis during PHA purification releases DNA, increasing medium viscosity and complicating downstream processing.
  • Current viscosity reduction methods (heat, hypochlorite, commercial nucleases) are costly.

Purpose of the Study:

  • To develop a cost-effective method for reducing viscosity in PHA downstream processing.
  • To engineer PHAs-producing bacteria with a nuclease gene for in-situ viscosity reduction.

Main Methods:

  • A nuclease gene from Staphylococcus aureus was integrated into Cupriavidus necator DSM 545 and Delftia acidovorans DSM 39.
  • The expression of staphylococcal nuclease in engineered microbial hosts was confirmed.
  • Viscosity measurements of cell lysates from recombinant and wild-type strains were compared.

Main Results:

  • Staphylococcal nuclease was successfully expressed in both C. necator and D. acidovorans without impacting PHA production.
  • The viscosity of lysates from recombinant C. necator cells was significantly reduced.
  • The engineered strains demonstrate potential for substantial cost reduction in PHA downstream processing.

Conclusions:

  • Engineering PHAs-producing bacteria with a Staphylococcus aureus nuclease gene is a viable strategy for reducing lysate viscosity.
  • This approach offers a more economical alternative to conventional methods for improving PHA purification efficiency.
  • The developed microbial strains are promising for large-scale, cost-effective industrial polyhydroxyalkanoates manufacturing.