De Novo Synthesis of Resveratrol from Sucrose by Metabolically Engineered Yarrowia lipolytica

Resveratrol, a phenylpropanoid compound, exhibits diverse pharmacological properties, making it a valuable candidate for health and disease management. However, the demand for resveratrol exceeds the capacity of plant extraction methods, necessitating alternative production strategies. Microbial synthesis offers several advantages over plant-based approaches and presents a promising alternative.Yarrowia lipolyticastands out among microbial hosts due to its safe nature, abundant acetyl-CoA and malonyl-CoA availability, and robust pentose phosphate pathway. This study aimed to engineerY. lipolyticafor resveratrol production. The resveratrol biosynthetic pathway was integrated intoY. lipolyticaby adding genes encoding tyrosine ammonia lyase fromRhodotorula glutinis, 4-coumarate CoA ligase fromNicotiana tabacum, and stilbene synthase fromVitis vinifera. This resulted in the production of 14.3 mg/L resveratrol. A combination of endogenous and exogenous malonyl-CoA biosynthetic modules was introduced to enhance malonyl-CoA availability. This included genes encoding acetyl-CoA carboxylase 2 fromArabidopsis thaliana, malonyl-CoA synthase, and a malonate transporter protein fromBradyrhizobium diazoefficiens. These strategies increased resveratrol production to 51.8 mg/L. The further optimization of fermentation conditions and the utilization of sucrose as an effective carbon source in YP media enhanced the resveratrol concentration to 141 mg/L in flask fermentation. By combining these strategies, we achieved a titer of 400 mg/L resveratrol in a controlled fed-batch bioreactor. These findings demonstrate the efficacy ofY. lipolyticaas a platform for the de novo production of resveratrol and highlight the importance of metabolic engineering, enhancing malonyl-CoA availability, and media optimization for improved resveratrol production.