Effective biosynthesis of 2,5-furandicarboxylic acid from 5-hydroxymethylfurfural via a bi-enzymatic cascade system using bacterial laccase and fungal alcohol oxidase.

Abstract

As a cost-effective and eco-friendly approach, biocatalysis has great potential for the transformation of 5-hydroxymethylfurfural (HMF) into 2,5-furandicarboxylic acid (FDCA). However, the compatibility of each enzyme in the cascade reaction limits the transformation efficiency of HMF to FDCA. Coupled with an alcohol oxidase from Colletotrichum gloeosporioides (CglAlcOx), this study aims to study the potential of bacterial laccase from Bacillus pumilus (BpLac) in an enzymatic cascade for 2,5-furandicarboxylic acid (FDCA) biosynthesis from 5-hydroxymethylfurfural (HMF). BpLac showed 100% selectivity for HMF oxidation and generated 5-hydroxymethyl-2-furancarboxylic acid (HMFCA). CglAlcOx was capable of oxidizing HMFCA to 2-formyl-5-furancarboxylic acid (FFCA). Both BpLac and CglAlcOx could oxidize FFCA to FDCA. At the 5mM scale, a complete transformation of HMF with a 97.5% yield of FDCA was achieved by coupling BpLac with CglAlcOx in the cascade reaction. The FDCA productivity in the reaction was 5.3mg/L/h. Notably, BpLac could alleviate the inhibitory effect of FFCA on CglAlcOx activity and boost the transformation efficiency of HMF to FDCA. Moreover, the reaction was scaled up to 40 times the volume, and FDCA titer reached 2.6mM with a yield of 58.77% at 168h. This work provides a candidate and novel insight for better design of an enzymatic cascade in FDCA production.