Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
70127 | Journal of Molecular Catalysis B: Enzymatic | 2012 | 5 Pages |
Feruloyl esterases (FAEs) are a group of important industrial enzymes, which could hydrolyze the ester bonds between hydroxycinnamic acids and arabinose or galactose in hemicellulose present in plant cell walls. To establish a practical high-throughput screening system of feruoyl esterases, two new substrates with modified chromo- or fluoro-phore, 2-chloro-4-nitrophenyl ferulate (CNPF; 1b) and umbelliferyl 5-O-feruolyl-α-L-arabinofuranoside (UFA; 2a), were designed and synthesized. Both substrates provided significantly improved signal-to-noise ratio compared with previously known structural analogues. The chromogenic substrate CNPF could be readily adapted to the high-throughput screening of feruloyl esterase A from Aspergillus niger, one of the most studied FAEs, with the coefficient of variance of as low as 10.3% as determined for a single sequence library.
Graphical abstractFigure optionsDownload full-size imageDownload as PowerPoint slideHighlights► Two new mimic substrates with modified chlorophore or fluorophore were synthesized. ► Each new substrate provides significantly improved signal-to-noise ratio. ► A high-throughput screening system was developed for feruloyl esterases.