DNA Plasmids for Production of Trans-cinnamic Acid

At a Glance

Researchers at Colorado State University have developed a novel method to produce trans-cinnamic acid (tCA) using engineered cyanobacteria. This technology offers a renewable and scalable alternative to fossil fuel-derived or plant-extracted tCA. Utilizing a rhamnose-inducible system, the engineered strain demonstrates up to a four-fold increase in tCA production. It advances sustainable bioproduction, addressing the growing demand for bioactive compounds in pharmaceuticals, cosmetics, and materials science.

Background

Trans-cinnamic acid (tCA) is a valuable precursor in pharmaceuticals, polymers, and cosmetics. Current production relies on fossil fuel-based synthesis or cost-prohibitive plant extraction. Cyanobacteria, capable of converting CO₂ and sunlight into valuable compounds, present a sustainable alternative. By engineering cyanobacteria to produce tCA efficiently, this technology provides a scalable and renewable solution to meet the rising global demand for high-value bioactive compounds.

Overview

This innovation leverages a model cyanobacterium to produce tCA using a plant-derived phenylalanine ammonia lyase (PAL) gene regulated by a rhamnose-inducible promoter. The engineered strain achieved up to 500 mg/L of tCA in 14 days under constant light in shake flasks, a four-fold improvement over previous studies. In environmental photobioreactors (ePBRs), the system exhibited 30-50% increased production efficiency under fluctuating light conditions compared to existing methods. This work validates the robustness and scalability of using inducible systems for bioproduction in cyanobacteria, setting the stage for industrial applications. Three constructs have been validated in both E. coli and a model cyanobacteria and are available for research only use from Addgene. Though the use and efficacy of these constructs has yet to be established specifically in additional species, the broad host range plasmid vector they are built on has been known to replicate in other species of Cyanobacteria and Pseudomonas, at a minimum.

Figure 1. trans-Cinnamic acid (tCA) production and growth of iNTP mutant shake flask cultures in 12- or 24-h light. (a) Average tCA production (mg L−1) under 24-h light (green) and 12:12 h light:dark cycles (blue) over 14 days. (b) Average production of tCA per culture density as approximated by optical density measured at 730 nm (mg L−1 · OD730−1). 50 mL S. 6803 iNTP mutant cultures were grown in shake flasks under 24-h light (green) or 12:12 hour light:dark cycles (blue) over 14 days. (c) Average culture growth of S. 6803 iNTP induced (purple), iNTP non-induced (teal), and empty vector-only control (yellow) as measured by optical density at 730 nm (OD730). 50 mL cultures were grown in shake flasks under either 12:12 h light:dark cycles (left) or 24-h light (right) over 14 days. Error bars indicate standard deviation of n = 3 biological replicates.

Benefits

• Renewable production: Converts CO₂ and sunlight into tCA, reducing reliance on fossil fuels.
• Improved efficiency: Up to four-fold higher tCA production compared to prior methods.
• Scalability: Demonstrated production in photobioreactors under realistic environmental conditions.
• Non-toxic induction: Utilizes a cost-effective, non-toxic rhamnose-inducible promoter.
• Versatility: Cross-species functionality demonstrated in both cyanobacteria and E. coli.

Applications

• Pharmaceutical precursors (e.g., anti-cancer, anti-viral, and anti-bacterial compounds).
• Cosmetic additives (e.g., fragrances, skin care products).
• Biodegradable polymers and plastics.
• Food additives (FDA-approved as Generally Regarded As Safe).
• Sustainable chemical manufacturing.

Publications

Hunstiger, et al (2024) “Improving trans-cinnamic acid production in a model cyanobacterium.” Biotechnology Progress. https://doi.org/10.1002/btpr.3512

Last Updated: August 2026

Opportunity

Material is available through Addgene.

IP Status

Biological Materials

Inventors

Christie Peebles
Hayley Ma
Darcy Hunstiger

Reference Number

2024-095

Licensing Manager

Jessy McGowan
Jessy.McGowan@colostate.edu
970-491-7100