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Heterologous production of a feruloyl esterase from Pleurotus sapidus synthesizing feruloyl‐saccharide esters
Author(s) -
Kelle Sebastian,
Nieter Annabel,
Krings Ulrich,
Zelena Katerina,
Linke Diana,
Berger Ralf G.
Publication year - 2015
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1002/bab.1430
Subject(s) - heterologous , esterase , biochemistry , chemistry , pectinase , enzyme , gene
Abstract The feruloyl esterase (FAE) gene EST1 from the basidiomycete Pleurotus sapidus was heterologously expressed in Escherichia coli and Pichia pastoris . Catalytically active recombinant Est1 was secreted using P. pastoris as a host. For expression in P. pastoris , the expression vector pPIC9K was applied. The EST1 gene was cloned with an N‐terminal α‐mating factor pre–pro sequence and expressed under the control of a methanol inducible alcohol oxidase 1 promotor. Est1 was purified to homogeneity using ion exchange and hydrophobic interaction chromatography. The recombinant Est1 showed optima at pH 5.0 and 50 °C, and released ferulic acid from saccharide esters and from the natural substrate destarched wheat bran. Substrate specificity profile and descriptor‐based analysis demonstrated unique properties, showing that Est1 did not fit into the current FAE classification model. Transferuloylation synthesis of feruloyl‐saccharide esters was proven for mono‐ and disaccharides.