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  1. Home
  3. Semi-automated Biofoundry Workflows For Sequence Coevolution-guided Isoprene Synthase Engineering.
  1. Home
  3. Semi-automated Biofoundry Workflows For Sequence Coevolution-guided Isoprene Synthase Engineering.

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Semi-automated biofoundry workflows for sequence coevolution-guided isoprene synthase engineering.

Georgii Emelianov1, Dong-Uk Song2, Aporva Kamath1

  • 1Synthetic Biology Research Center and Korea Biofoundry, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 34141, Republic of Korea; Department of Biosystems and Bioengineering, KRIBB School of Biotechnology, University of Science and Technology (UST), Daejeon 34113, Republic of Korea.

Trends in Biotechnology
|September 13, 2025

View abstract on PubMed

Summary
This summary is machine-generated.

Biofoundries accelerate biomanufacturing through scalable enzyme engineering. This study enhanced isoprene synthase (IspS) activity and stability, improving methane-to-isoprene bioconversion by 4.5-fold.

Keywords:
biofoundryenzyme engineeringisopreneisoprene synthase

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Area of Science:

  • Biotechnology and Synthetic Biology
  • Enzyme Engineering
  • Biomanufacturing

Background:

  • Biofoundries are crucial for advancing enzyme and microorganism engineering.
  • Isoprene synthase (IspS) is a key enzyme in isoprene biosynthesis, often acting as a rate-limiting step.
  • Efficient enzyme engineering is vital for optimizing biomanufacturing processes.

Purpose of the Study:

  • To develop scalable enzyme engineering workflows for biofoundry applications.
  • To improve the catalytic efficiency and thermostability of isoprene synthase (IspS).
  • To enhance methane-to-isoprene bioconversion using engineered IspS in Methylococcus capsulatus Bath.

Main Methods:

  • Integrated computational mutation design using sequence coevolution analysis.
  • Employed laboratory automation for high-throughput site-directed mutagenesis and screening.
  • Conducted three rounds of genetic mutant synthesis and evaluation, demonstrating scalability to thousands of mutants.

    • Main Results:

    • Identified IspS variants with up to a 4.5-fold increase in catalytic efficiency.
    • Achieved enhanced thermostability in engineered IspS variants.
    • Improved methane-to-isoprene bioconversion in Methylococcus capsulatus Bath to a titer of 319.6 mg/l.

    Conclusions:

    • Developed robust and scalable enzyme engineering workflows suitable for biofoundries.
    • Demonstrated significant improvements in IspS function and bioconversion efficiency.
    • Established a framework for future biotechnological advancements through enzyme engineering.