title: A feruloyl esterase (FAE) characterized by relatively high thermostability from the edible mushroom Russula virescens.
abstract: A monomeric feruloyl esterase (FAE) with a molecular mass of 62 kDa was acquired from fresh fruiting bodies of the edible mushroom Russula virescens. The isolation procedure involved ion exchange chromatography on CM-cellulose, Q-Sepharose, and SP-Sepharose and finally fast protein liquid chromatography-gel filtration on Superdex 75. Two amino acid sequences were obtained after tryptic digestion, and they both showed some homology with the esterase of some fungi. Maximal activity was observed at pH 5.0 and at 50 °C. The enzyme displayed relatively high thermostability as evidenced by over 70 % residual activity at 70 °C and about 34 % residual activity at 80 °C. The K m and V max for this enzyme on methyl ferulate were 0.19 mM and 1.65 U/mg proteins, respectively. The purified FAE prefers methyl ferulate over methyl caffeate and is least active on methyl p-coumarate. The FAE activity was not significantly affected by the presence of cations such as Mn(2+), Ca(2+), Cd(2+), Zn(2+), Mg(2+), Cu(2+), and K(+) ions but inhibited by Al(3+), Hg(2+), Fe(2+), and Pb(2+) ions at a tested concentration of 2. 5 mM.
authors.affiliation: State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University, Beijing, 100193, China.
authors.forename: Li
authors.initials: L
authors.lastname: Wang
authors.forename: Rui
authors.initials: R
authors.lastname: Zhang
authors.forename: Zengqiang
authors.initials: Z
authors.lastname: Ma
authors.forename: Hexiang
authors.initials: H
authors.lastname: Wang
authors.forename: Tzibun
authors.initials: T
authors.lastname: Ng
mesh_terms: D000363:Agaricales / Q000187:drug effects / Q000201:enzymology*; D000595:Amino Acid Sequence; D002265:Carboxylic Ester Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003373:Coumaric Acids / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability / Q000187:drug effects; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D007477:Ions; D007700:Kinetics; D008670:Metals / Q000494:pharmacology; D008970:Molecular Weight; D013379:Substrate Specificity / Q000187:drug effects; D013696:Temperature
chemical_list: D003373:Coumaric Acids; D007477:Ions; D008670:Metals; C004999:ferulic acid; D002265:Carboxylic Ester Hydrolases; C070196:feruloyl esterase