Metabolic engineering of Escherichia coli for upcycling of polyethylene terephthalate waste to vanillin

Polyethylene terephthalate (PET) waste presents a significant environmental challenge due to its durability and resistance to degradation. Innovative approaches for upcycling PET waste into high-value chemicals can mitigate these issues while contributing to a circular economy. In this study, we developed a multi-enzyme cascade system in E. coli to convert PET-derived monomer terephthalic acid (TPA) into vanillin (VAN). The metabolic engineering approach was then employed to increase VAN production, including 1) inhibition of VAN degradation by knocking out endogenous aldehyde reductases and alcohol dehydrogenases and 2) enhancement of TPA uptake by modifying membrane proteins to increase cell permeability. The engineered E. coli demonstrated a VAN production of 658.55 mg/L from 1992 mg/L of TPA with a molar conversion rate of 71.1 %, representing the highest production of VAN using TPA as the substrate. Additionally, the engineered E. coli effectively converted post-consumer PET waste into VAN under mild conditions, with the highest production of 259.2 mg/L in 20× diluted PET hydrolysates, highlighting its potential for application in PET waste upcycling. This approach not only provides an environmentally friendly alternative to traditional chemical synthesis but also offers substantial economic benefits by transforming low-value waste into high-value chemicals.