Prenylated stilbenoids possess a range of biological activities, including anti-inflammatory, antioxidant, antidiabetic, anticancer, neuroprotective and vasorelaxant properties. However, their potential is limited by the scarcity of these compounds in nature. In this study, we present an eco-friendly biosynthetic method for producing prenylated resveratrol in Escherichia coli. By co-expressing the MVA pathway for DMAPP synthesis with a prenyltransferase, we successfully achieved in vivo prenylation of resveratrol. We then reconstructed a novel biosynthetic pathway for 2-C-prenyl resveratrol by integrating the resveratrol pathway. Through careful metabolic flux engineering to accumulate acetyl-CoA and malonyl-CoA, we created a high-yield strain that produced 2-C-prenyl resveratrol, achieving a significant titer of 68.4 mg L−1 in flask cultures. This research marks the first microbial production of prenylated stilbenoids, providing a foundation for sustainable green production of these compounds.
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