Biosynthesis of pterostilbene in <i>Escherichia coli</i> from resveratrol on macroporous adsorption resin using a two‐step substrate addition strategy

Pterostilbene (PST), a 3',5'-O-methylated derivative of resveratrol (RSV), is a potent natural antioxidant produced by some plants in trace amounts as defense compound. It exhibits various health-promoting activities, such as anticancer, antiviral, and antimicrobial effects. Large-scale biosynthesis of PST is crucial due to the challenges associated with extracting it from plants. This study aims to develop an efficient method for PST production using an engineered Escherichia coli strain by feeding RSV as a precursor. We introduced a two-step substrate addition strategy combined with immobilized RSV (IMRSV) on macroporous adsorption resin (MAR) to enhance PST production. Five MARs were selected for RSV immobilization, and the substrate addition strategy and fermentation parameters for PST synthesis were optimized. A maximum PST concentration of 403 ± 9 mg/L was achieved, representing a 239% increase over the control, which in a one-step addition of free RSV. The PST titer reached 395 ± 24 mg/L in a 3-L bioreactor. In conclusion, the combination of a two-step substrate addition system and IMRSV is a promising approach for the economical and industrial-scale production of PST.