Lignin as a byproduct can be obtained
from agricultural wastes
and biorefinery. In this study, we sought to develop Escherichia
coli-based whole-cell biocatalysis for converting lignin-derived
isoeugenol into cis,cis-muconic acid (ccMA). Recombinant E. coli expressing aromatic dioxygenase from Thermothelomyces
thermophila and aldehyde dehydrogenase from Saccharomyces
cerevisiae could efficiently convert isoeugenol to vanillic
acid (VAN). However, we found that the demethylation of VAN by VanAB
from Pseudomonas putida was severely inhibited by
a high concentration of isoeugenol, which prompted us to develop a
two-step approach to the production of ccMA from isoeugenol. Upon
adjustment of the feeding strategy and optimization of the pH conditions,
43.8 mM ccMA was produced from 50 mM isoeugenol-derived VAN by recombinant E. coli expressing VanAB from P. putida, GDC from Talaromyces atroroseus, and CatA from Acinetobacter radioresistens. This work is a pioneering
study in the evaluation of E. coli-based whole-cell
biocatalysis for lignin valorization.