title: Substrate and positional specificity of feruloyl esterases for monoferuloylated and monoacetylated 4-nitrophenyl glycosides.
abstract: 4-Nitrophenyl glycosides of 2-, 3-, and 5-O-(E)-feruloyl- and 2- and 5-O-acetyl-alpha-L-arabinofuranosides and of 2-, 3-, and 4-O-(E)-feruloyl- and 2-, 3- and 4-O-acetyl-beta-D-xylopyranosides, compounds mimicking natural substrates, were used to investigate substrate and positional specificity of type-A, -B, and -C feruloyl esterases. All the feruloyl esterases behave as true feruloyl esterases showing negligible activity on sugar acetates. Type-A enzymes, represented by AnFaeA from Aspergillus niger and FoFaeII from Fusarium oxysporum, are specialized for deferuloylation of primary hydroxyl groups, with a very strong preference for hydrolyzing 5-O-feruloyl-alpha-L-arabinofuranoside. On the contrary, type-B and -C feruloyl esterases, represented by FoFaeI from F. oxysporum and TsFaeC from Talaromyces stipitatus, acted on almost all ferulates with exception of 4- and 3-O-feruloyl-beta-D-xylopyranoside. 5-O-Feruloyl-alpha-L-arabinofuranoside was the best substrate for both TsFaeC and FoFaeI, although catalytic efficiency of the latter enzyme toward 2-O-feruloyl-alpha-L-arabinofuranoside was comparable. In comparison with acetates, the corresponding ferulates served as poor substrates for the carbohydrate esterase family 1 feruloyl esterase from Aspergillus oryzae. The enzyme hydrolyzed all alpha-L-arabinofuranoside and beta-D-xylopyranoside acetates. It behaved as a non-specific acetyl esterase rather than a feruloyl esterase, with a preference for 2-O-acetyl-beta-D-xylopyranoside.
authors.affiliation: Institute of Chemistry, Slovak Academy of Sciences, Dúbravská cesta 9, SK-845 38 Bratislava, Slovak Republic.
authors.forename: Vladimír
authors.initials: V
authors.lastname: Puchart
authors.forename: Mária
authors.initials: M
authors.lastname: Vrsanská
authors.forename: Mária
authors.initials: M
authors.lastname: Mastihubová
authors.forename: Evangelos
authors.initials: E
authors.lastname: Topakas
authors.forename: Christina
authors.initials: C
authors.lastname: Vafiadi
authors.forename: Craig B
authors.initials: CB
authors.lastname: Faulds
authors.forename: Maija
authors.initials: M
authors.lastname: Tenkanen
authors.forename: Paul
authors.initials: P
authors.lastname: Christakopoulos
authors.forename: Peter
authors.initials: P
authors.lastname: Biely
mesh_terms: D000107:Acetylation; D000115:Acetylesterase / Q000378:metabolism; D001089:Arabinose / Q000737:chemistry / Q000378:metabolism; D001234:Aspergillus niger / Q000201:enzymology; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D005658:Fungi / Q000201:enzymology*; D005670:Fusarium / Q000201:enzymology; D006026:Glycoside Hydrolases / Q000378:metabolism; D006027:Glycosides / Q000737:chemistry / Q000378:metabolism*; D006868:Hydrolysis; D007700:Kinetics; D013379:Substrate Specificity; D032901:Talaromyces / Q000201:enzymology
chemical_list: D006027:Glycosides; D001089:Arabinose; D002265:Carboxylic Ester Hydrolases; D000115:Acetylesterase; C070196:feruloyl esterase; D006026:Glycoside Hydrolases; C019713:alpha-N-arabinofuranosidase