Multi-enzymatic cascade reactions with Escherichia coli-based modules for synthesizing various bioplastic monomers from fatty acid methyl esters

Multi-enzymatic cascade reaction systems were designed to generate biopolymer monomers using Escherichia coli-based cell modules, capable of carrying out one-pot reactions. Three cell-based modules, including a omega-hydroxylation module (Cell-H-m) to convert fatty acid methyl esters (FAMEs) to omega-hydroxy fatty acids (omega-HFAs), an amination module (Cell-A(m)) to convert terminal alcohol groups of the substrate to amine groups, and a reduction module (Cell-R-m) to convert the carboxyl groups of fatty acids to alcohol groups, were constructed. The product-oriented assembly of these cell modules involving multi-enzymatic cascade reactions generated omega-ADAs (up to 46 mM), alpha,omega-diols (up to 29 mM), omega-amino alcohols (up to 29 mM) and alpha,omega-diamines (up to 21 mM) from 100 mM corresponding FAME substrates with varying carbon chain length (C8, C10, and C12). Finally 12-ADA and 1,12-diol were purified with isolated yields of 66.5% and 52.5%, respectively. The multi-enzymatic cascade reactions reported herein present an elegant 'greener' alternative for the biosynthesis of various biopolymer monomers from renewable saturated fatty acids.N