Heterologous production of a feruloyl esterase from Pleurotus sapidus synthesizing feruloyl‐saccharide esters

Sebastian Kelle; Annabel Nieter; Ulrich Krings; Katerina Zelena; Diana Linke; Ralf G. Berger

doi:10.1002/bab.1430

journal article Oct 20, 2015

Heterologous production of a feruloyl esterase from Pleurotus sapidus synthesizing feruloyl‐saccharide esters

Sebastian Kelle

Annabel Nieter Ulrich Krings Katerina Zelena Diana Linke Ralf G. Berger

Biotechnology and Applied Biochemistry Vol. 63 No. 6 pp. 852-862 · Wiley

View at Publisher Save 10.1002/bab.1430

Abstract

AbstractThe 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.

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Details

Published: Oct 20, 2015
Vol/Issue: 63(6)
Pages: 852-862
License: View

Authors

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Sebastian Kelle