Abstract
(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 related pathway intermediate p-coumaric acid, which is an important factor for efficient production. Here, we show that fine-turning of p-coumaric acid synthesis enables alleviated cell burden and improved (2S)-naringenin production in yeast. First, we systematically 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 amounts of p-coumaric acid still accumulated outside the cell. We further established a dual dynamic control system through combing a malonyl-CoA biosensor regulator and an RNAi strategy, to autonomously control the synthesis of p-coumaric acid and downregulate a pathway competing for malonyl-CoA. The optimized strains remarkably decreased extracellular accumulation of p-coumaric acid and simultaneously improved (2S)- naringenin production. Finally, production of 933 mg/L of (2S)-naringenin could be achieved by using minimal medium with negligible accumulation of p-coumaric acid. Our work highlights the importance of systematic control of pathway intermediates for efficient microbial production of plant natural products.
Competing Interest Statement
The authors have declared no competing interest.
