title: A halotolerant type A feruloyl esterase from Pleurotus eryngii.
abstract: An extracellular feruloyl esterase (PeFaeA) from the culture supernatant of Pleurotus eryngii was purified to homogeneity using cation exchange, hydrophobic interaction, and size exclusion chromatography. The length of the complete coding sequence of PeFaeA was determined to 1668 bp corresponding to a protein of 555 amino acids. The catalytic triad of Ser-Glu-His demonstrated the uniqueness of the enzyme compared to previously published FAEs. The purified PeFaeA was a monomer with an estimated molecular mass of 67 kDa. Maximum feruloyl esterase (FAE) activity was observed at pH 5.0 and 50 °C, respectively. Metal ions (5 mM), except Hg(2+), had no significant influence on the enzyme activity. Substrate specificity profiling characterized the enzyme as a type A FAE preferring bulky natural substrates, such as feruloylated saccharides, rather than small synthetic ones. Km and kcat of the purified enzyme for methyl ferulate were 0.15 mM and 0.85 s(-1). In the presence of 3 M NaCl activity of the enzyme increased by 28 %. PeFaeA alone released only little ferulic acid from destarched wheat bran (DSWB), whereas after addition of Trichoderma viride xylanase the concentration increased more than 20 fold.
authors.affiliation: Institut für Lebensmittelchemie, Leibniz Universität Hannover, Callinstraße 5, D-30167 Hannover, Germany. Electronic address: annabel.nieter@lci.uni-hannover.de.
authors.forename: Annabel
authors.initials: A
authors.lastname: Nieter
authors.affiliation: Institut für Lebensmittelchemie, Leibniz Universität Hannover, Callinstraße 5, D-30167 Hannover, Germany.
authors.forename: Paul
authors.initials: P
authors.lastname: Haase-Aschoff
authors.affiliation: Institut für Lebensmittelchemie, Leibniz Universität Hannover, Callinstraße 5, D-30167 Hannover, Germany.
authors.forename: Diana
authors.initials: D
authors.lastname: Linke
authors.affiliation: Helmholtz Zentrum für Infektionsforschung, Inhoffenstraße 7, D-38124 Braunschweig, Germany.
authors.forename: Manfred
authors.initials: M
authors.lastname: Nimtz
authors.affiliation: Institut für Lebensmittelchemie, Leibniz Universität Hannover, Callinstraße 5, D-30167 Hannover, Germany.
authors.forename: Ralf G
authors.initials: RG
authors.lastname: Berger
mesh_terms: D002265:Carboxylic Ester Hydrolases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D002853:Chromatography, Liquid; D003373:Coumaric Acids / Q000378:metabolism; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D004043:Dietary Fiber / Q000378:metabolism; D020536:Enzyme Activators / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000378:metabolism; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008670:Metals / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D020076:Pleurotus / Q000201:enzymology*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012965:Sodium Chloride / Q000378:metabolism; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D003373:Coumaric Acids; D004271:DNA, Fungal; D004043:Dietary Fiber; D020536:Enzyme Activators; D004791:Enzyme Inhibitors; D008670:Metals; D012965:Sodium Chloride; C004999:ferulic acid; D002265:Carboxylic Ester Hydrolases; C070196:feruloyl esterase
keywords: Basidiomycete; Ferulic acid esterase; Feruloyl saccharides; Fungi; Halotolerance