Fine-tuning of p-coumaric acid synthesis to increase (2S)-naringenin production in yeast.

Mao J, Mohedano MT, Fu J, Li X, Liu Q, Nielsen J, Siewers V, Chen Y

Metab. Eng. 79 (-) 192-202 [2023-09-00; online 2023-08-22]

(2S)-Naringenin is a key precursor for biosynthesis of various high-value flavonoids and possesses a variety of nutritional and pharmaceutical properties on human health. Systematic optimization approaches have been employed to improve (2S)-naringenin production in different microbial hosts. However, very few studies have focused on the spatiotemporal distribution of (2S)-naringenin and the related pathway intermediate p-coumaric acid, which is an important factor for efficient production. Here, we first optimized the (2S)-naringenin biosynthetic pathway by alleviating the bottleneck downstream of p-coumaric acid and increasing malonyl-CoA supply, which improved (2S)-naringenin production but significant accumulation of p-coumaric acid still existed extracellularly. We thus established a dual dynamic control system through combining a malonyl-CoA biosensor regulator and an RNAi strategy, to autonomously control the synthesis of p-coumaric acid with the supply of malonyl-CoA. Furthermore, screening potential transporters led to identification of Pdr12 for improved (2S)-naringenin production and reduced accumulation of p-coumaric acid. Finally, a titer of 2.05 g/L (2S)-naringenin with negligible accumulation of p-coumaric acid was achieved in a fed batch fermentation. Our work highlights the importance of systematic control of pathway intermediates for efficient microbial production of plant natural products.

Chalmers Mass Spectrometry Infrastructure [Service]

PubMed 37611820

DOI 10.1016/j.ymben.2023.08.003

Crossref 10.1016/j.ymben.2023.08.003

pii: S1096-7176(23)00117-9

Publications 9.5.0