Metabolic engineering of Escherichia coli for production of acetaminophen from glucose based on high selectivity of N-acetyltransferase

Acetaminophen, also known as paracetamol, is one of the most commonly used medicines and is widely accepted as a safe and effective analgesic/antipyretic for mild to moderate pain and fever. Several efforts have been made on engineered E. coli for the biosynthesis of acetaminophen. However, the yield of de novo biosynthesis of acetaminophen from glucose still needs to be improved. In this study, we identified an N-acetyltransferase svNAT2 from Streptomyces venezuelae FBUA 1571with high substrate specificity for p-aminophenol. Subsequently, based on this N-acetyltransferase, we constructed a biosynthetic pathway for acetaminophen from glucose in E. coli. The results show that using high selectivity of N-acetyltransferase svNAT2 can effectively improve the production of acetaminophen. Subsequently, we further increased the acetaminophen production to 1268 mg/L by the increasing availability of erythrose-4-phosphate and phosphoenolpyruvate.