# Engineering a Carboxyl Methyltransferase for the Formation of a Furan‐Based Bioplastic Precursor

## TL;DR

Scientists engineered a new enzyme that efficiently converts a bio-based acid into a bioplastic precursor, enabling sustainable plastic production and recycling.

## Contribution

A rationally designed FtpM mutant (R166M) enables high-efficiency dimethylation of FDCA for bioplastic precursor synthesis.

## Key findings

- The R166M mutant achieves near-quantitative conversion of FDCA to FDME.
- The mutant was integrated into two enzymatic cascades for FDME synthesis from HMF or PEF plastic.
- The mutant shows improved activity for various diacids and monoacids.

## Abstract

FtpM from Aspergillus fumigatus was the first carboxyl methyltransferase reported to catalyse the dimethylation of dicarboxylic acids. Here the creation of mutant R166M that can catalyse the quantitative conversion of bio‐derived 2,5‐furandicarboxylic acid (FDCA) to its dimethyl ester (FDME), a bioplastics precursor, was reported. Wild type FtpM gave low conversion due to its reduced catalytic efficiency for the second methylation step. An AlphaFold 2 model revealed a highly electropositive active site, due to the presence of 4 arginine residues, postulated to favour the binding of the dicarboxylic acid over the intermediate monoester. The R166M mutation improved both binding and turnover of the monoester to permit near quantitative conversion to the target dimethyl ester product. The mutant also had improved activity for other diacids and a range of monoacids. R166M was incorporated into 2 multienzyme cascades for the synthesis of the bioplastics precursor FDME from bioderived 5‐hydroxymethylfurfural (HMF) as well as from poly(ethylene furanoate) (PEF) plastic, demonstrating the potential to recycle waste plastic.

Rationally designed carboxyl methyltransferase FtpM mutant R166M catalyses the dimethylation of 2,5‐furandicarboxylic acid (FDCA) in very high conversion. FtpM R166M was integrated into two different one‐pot enzymatic cascades for conversion of bioderived 5‐hydroxymethylfurfural or poly(ethylene furanoate) plastic to FDCA dimethyl ester (FDME), using oxidases or cutinase respectively.