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Cell-free styrene biosynthesis at high titers.

William S Grubbe1, Blake J Rasor1, Antje Krüger1

  • 1Department of Chemical and Biological Engineering, Northwestern University, Evanston, IL, 60208, USA; Chemistry of Life Processes Institute, Northwestern University, Evanston, IL, 60208, USA; Center for Synthetic Biology, Northwestern University, Evanston, IL, 60208, USA.

Metabolic Engineering
|June 6, 2020
PubMed
Summary

Developing a cell-free system for styrene biosynthesis overcomes limitations of cellular production. This method enhances styrene yield, offering a sustainable alternative to petroleum-based plastics.

Keywords:
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Area of Science:

  • Biotechnology
  • Chemical Engineering
  • Synthetic Biology

Background:

  • Styrene is a key petroleum-derived monomer for plastics, facing sustainability challenges due to fossil fuel dependence and climate change.
  • Current biological styrene production methods in engineered *Escherichia coli* are hindered by product toxicity and volatility.
  • Developing sustainable, bio-based routes for styrene is crucial for reducing reliance on fossil fuels.

Purpose of the Study:

  • To engineer a cell-free biosynthesis platform for styrene production.
  • To overcome the limitations of cellular toxicity and volatility in biological styrene synthesis.
  • To optimize reaction conditions for maximizing styrene titer in a cell-free system.

Main Methods:

  • Utilized cell-free protein synthesis to generate the two required biosynthetic enzymes for styrene.
  • Combined enzymes with L-phenylalanine and buffer in an open reaction system.
  • Optimized reaction parameters including time, temperature, pH, and enzyme concentrations.

Main Results:

  • Achieved a significant increase in cell-free styrene titer, from 5.36 ± 0.63 mM to 40.33 ± 1.03 mM.
  • Demonstrated the highest styrene yield via biosynthesis without product removal or process modifications.
  • Established a robust cell-free platform for styrene production.

Conclusions:

  • Cell-free systems provide an advantageous alternative to cellular biosynthesis for producing toxic and volatile molecules like styrene.
  • This platform offers enhanced control over reaction conditions and improved carbon flux towards product formation.
  • The developed cell-free system represents a significant advancement in sustainable styrene production methods.