Implementing a Design, Build, Test, Learn P. kudriavzevii engineering cycle for production of an organic acid product

Lygos is an industrial biotechnology company spun out of the U.S. Department of Energy-funded Joint BioEnergy Institute. Lygos uses Pichia kudriavzevii, an acid tolerant BSL-1 yeast, to produce organic acids, chemicals that are generally expensive to manufacture petrochemically but that can be produced at high yields and for low cost biologically.  While P. kudriavzevii’s high tolerance for organic acids provides an advantage over E. coli, S. cerevisiae, and many other microbes, working with P. kudriavzevii presents unique challenges. For example, there is little published transcript and protein expression data for this organism, which could be used to guide strain engineering or to build new genetic tools. While Lygos has developed protocols and tools for designing, building, and testing P. kudriavzevii strains, there remains room to improve the engineering cycle, providing an opportunity to partner with the Agile BioFoundry to further advance this microorganism.

This project has at least two high-level goals.

  • Apply the Agile BioFoundry DBTL tools and technologies to establish and optimize production of a new, valuable, organic acid product in P. kudriavzevii
  • Onboard P. kudriavzevii as a production host within the Agile BioFoundry.

The organic acid selected for development is a commercial product that is difficult and costly to make petrochemically. As an added benefit, this organic acid is also a metabolic intermediate useful for the production of other commercially important chemicals, making it an ideal candidate to demonstrate the commercial applicability of Agile BioFoundry DBTL technologies.

Relevant ABF Capabilities:

Project Partners:
Lygos, SNL, LBNL, and PNNL

Jeffrey Dietrich (Lygos), John Gladden (SNL), Jon Magnuson (PNNL), Taek Soon Lee (LBNL)

Performance period: 04/2019 to 03/2021