Published Feb. 11, 2026

test tubes marked biofuel

Applications

  • Development of modular workflows to build new genetic circuits as biosensors 
  • Demonstrated improvement to jet-fuel precursors, like isoprenol, using P. putida
  • High-throughput strain engineering for improved bioproduct yields
  • Biosensor-based selection platform for optimizing metabolic pathways 

Advantages/Benefits

  • Increased isoprenol titers 36-fold using rewired Pseudomonas putida strains
  • Adaptive, versatile platform to other biosensors/metabolic pathways in P. putida
  • Enables discovery of new genetic targets through unbiased, high-throughput methods

Background

Isoprenol (3-methyl-3-buten-1-ol CAS: 763-32-6) is an important building block for sustainable aviation fuel and a beachhead molecule for many commodity chemicals. However, traditional methods that optimize microbial strains for high-yield and efficient microbial production typically require time-intensive analytical methods, like gas chromatography. This technology addresses that challenge by linking isoprenol production directly to cellular growth through a biosensor-based genetic circuit, enabling rapid and scalable screening of beneficial genetic mutations.

Technology Overview

Researchers at Berkeley Lab have developed a novel strain engineering platform using a biosensor that helps bacteria grow and activate the expression of an essential gene only in the presence of isoprenol. By testing a CRISPRi gRNA library (with 16,000 P. putida gene targets) introduced into P. putida, scientists quickly identified genetic changes that have enhanced isoprenol production from 25 mg/L to 900 mg/L, which is about a thirty-five fold improvement in production. Strains with increased growth (and thus increased isoprenol output) were regenerated with recombineering using an iterative, data-driven process, and their improved titers were validated by gas chromatography analysis. This combined approach, linking growth to fuel production and genome-scale screening, revealed previously unknown genetic targets and significantly boosted biofuel precursor production.

Development Stage

Proof of Concept; TRL between 3-4

For More Information:

Menasalvas, J. et al. 24 Oct 2025. Biosensor-driven strain engineering reveals key cellular processes for maximizing isoprenol production in Pseudomonas putida. Science Advances, Vol 11, Issue 43. https://www.science.org/doi/10.1126/sciadv.ady2677

Inventors:

  • Thomas T. Eng
  • Javier Menasalvas
  • Aindrila Mukhopadhyay

Status

Patent pending 

Opportunities

Available for licensing or collaborative research