Design of chromogenic probes for efficient screening and evaluation of feruloyl esterase-like activities
Abstract
Feruloyl esterases (FAEs) constitute an important sub-group of hydrolytic enzymes involved in the deconstruction of plant cell wall polysaccharides. However, in the current era of genomics and metagenomics, finding and characterizing FAEs is not straightforward, mainly due to a lack of suitable compounds for high-throughput assays. To remedy this, indolyl and 4-nitrocatechol hydroxycinnamates (i.e. transferulate and p-coumarate derivatives) were synthesized in good overall yields-between 46 and 56% after 4 steps- and their usefulness as substrates for FAEs was ascertained. The hydrolysis of the ester bond of these chromogenic compounds leads to a colour change, which can be readily monitored. Overall, these compounds considerably improve upon the current situation and enable the measurement of FAE activities in both qualitative solid medium-based and quantitative liquid assays.