Industrial production of natural low-calorie sweetener neohesperidin dihydrochalcone

Neohesperidin dihydrochalcone (NHDC), a natural, low-calorie, high-intensity sweetener derived from plants, faces sustainable supply challenges due to its limited natural abundance. Here we developed a biochemical synthetic process for NHDC production at 5000-L bioreactor system. First, a heterologous synthetic pathway for neohesperidin, which is the precursor of NHDC, was constructed in Yarrowia lipolytica through the screening and assembly of candidate genes sourced from 25 species. Through key gene mining, metabolic flux regulation and fermentation optimization, we constructed 106 engineering strains and improved the titers of neohesperidin by 6,481-fold reaching 28.52 g/L. Finally, the precursor neohesperidin was integrated with a chemical conversion system leading to the establishment of an efficient biochemical process for high-yield NHDC production at an industrial scale. Our findings provide a roadmap for sustainable production of high-value flavonoids and their derivatives.