Applications

• Enhanced metabolism and expression of complex metabolites in plants
• Improved yields of complex molecules in target host cells 
• Production of valuable compounds, such as pharmaceuticals and biofuels

Advantages/Benefits

• Increased transformation efficiency and target molecule production 
• Predictive quantitative model to aid in experimental design and metabolic engineering 
• Scalable production of diverse genetic pathways 

Background

In recent years, advancements in biotechnology platforms have helped to improve metabolic engineering in plants, offering potential opportunities for production of useful compounds. Agroinfiltration is one such method that temporarily expresses plant genes and specific proteins, widely used in basic and applied plant science. Competition among individual bacteria often poses limitations on the genetic complexity achievable in plants and new systems must be utilized to overcome this and allow for improved metabolic production of genetically complex pathways. 

Technology Overview

Researchers at the Joint BioEnergy Institute have introduced a system to combat competition between Agrobacterium strains and thus improve the titer of target molecules. They engineered bacterial strains, termed “BiBi” strains, using Agrobacterium with two binary vectors (pVS1 and BBR1) to enhance efficiency of plant metabolism in Nicotiana benthamiana (Tobacco) leaf cells. The implementation of the BiBi system demonstrated a significant improvement in plant cell transformation efficiency, with 44% of epidermis nuclei exhibiting detectable levels of green or red fluorescent protein (GFP, RFP) expression, surpassing conventional methods. They also introduced a Poisson model framework, previously unused in agroinfiltration, to assess performance, bacterial concentration, and the likelihood of plant cell infection. 

By co-delivering multiple T-DNAs from a single strain, BiBi bypassed bacterial competition, particularly evident at lower bacterial densities. This breakthrough not only showcases the BiBi system’s ability to synthesize complex metabolites, but also highlights its value in commercial applications and the field of plant metabolic engineering. 

Development Stage

Proof of concept

Principal Investigator(s)

• Mitchell G. Thompson 
• Patrick M. Shih 
• Simon Alamos 
• Matthew Szarzanowicz 
• Liam Kirkpatrick 

Status

Patent pending

Opportunities

Available for licensing or collaborative research