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. They have been widely applied to biomass degradation, pulping, food and pharmaceutical industries. Recently, feruloyl esterases have attracted much attention of researchers. To facilitate mining novel enzyme mining from environmental microorganisms and the directed evolution of FAEs, an efficient high-throughput screening system was demanded.
Prof. WU Zhongliu and coworkers from Chengdu Institute of Biology, Chinese Academy of Sciences established a practical high-throughput screening system of feruoyl esterases, two new substrates with modified chromo- or fluoro-phore, 2-chloro-4-nitrophenyl ferulate and umbelliferyl 5- O-feruolyl-a-L-arabinofuranoside, 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.
WU’s research entitled “A practical high-throughput screening system for feruloyl esterases: Substrate design and evaluation” has been published in Journal of Molecular Catalysis B: Enzymatic (74 (2012) 36– 40)..This work was supported by the Program of 100 Distinguished Young Scientists and the Knowledge Innovation Program of the Chinese Academy of Sciences (No. KSCX1-YW-11B2), the National Natural Science Foundation of China (20802073 and 21072183), the Province Science Foundation of Sichuan, China (Nos. 08ZQ026-023 and 2010SZ0128).
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