Engineering of escherichia coli for high-level production of p-coumaric acid from glucose

Abstract

p-Coumaric acid is industrial desirable plant origin phenolic acid having anti-oxidant and anti-inflammatory effects, which can be used as a precursor for various valuable flavonoids. p-Coumaric acid successfully biosynthesized from glucose by co-expressing Arabidopsis thaliana origin trans-cinnamate 4-hydroxylase (C4H) and cytochrome P450 reductase (CPR) in trans-cinnamic acid producing Escherichia coli strain. With culture temperature and media optimization, up to 484 mg/L of titer was achieved in fed-batch cultivation. To increase the activity of heterologous C4H in bacteria, directed evolution of enzyme approach was additionally introduced. p-Coumaric acid specific bacterial biosensor was used for the high-throughput fluorescence-based enzyme library screening and capable of isolating mutations associated with increased p-coumaric acid titer.