Development of a Komagataella phaffii cell factory for sustainable production of ( +)-valencene.

Summary

Using CRISPR/Cas9, enzyme fusion, pathway flux enhancement, promoter modulation, and gene copy optimization, the authors engineered K. phaffii to produce (+)-valencene at 173.6 mg/L (82-fold over the starting strain). This advances sustainable fragrance supply for food, beverage, and cosmetics with a modular strategy generalizable to other terpenoids.

Key Findings

• CRISPR/Cas9-enabled introduction of (+)-valencene synthase yielded an initial 2.1 mg/L producer strain.
• Fusion of farnesyl pyrophosphate synthase to valencene synthase increased titers to 8.2 mg/L; overexpression of IDI1, tHMG1, ERG12, and ERG19 further boosted yield by 27%.
• Promoter deletion of ERG9 and optimization to three copies of the fusion construct achieved 173.6 mg/L in shake flasks, an 82-fold increase over the starting strain.

Clinical Implications

While not a clinical trial, this work can stabilize fragrance ingredient supply chains for dermatologic and cosmetic formulations, potentially improving consistency, cost, and sustainability of topical products.

Limitations

• Results are at shake-flask scale; bioreactor optimization and downstream processing not reported.
• Productivity, yield on substrate, and cost models versus plant extraction are not analyzed.

Future Directions

Scale-up in bioreactors, process intensification, pathway balancing for higher yields, and techno-economic/life-cycle analyses compared with agricultural sourcing.