Purification and partial characterization of two feruloyl esterases from the anaerobic fungus Neocallimastix strain MC-2.
Two extracellular feruloyl esterases (FAE-I and FAE-II) produced by the anaerobic fungus Neocallimastix strain MC-2 which cleave ferulic acid from O-(5-O-[(E)-feruloyl]-alpha-L- arabinofuranosyl)-(1-->3)-O-beta-D-xylopyranosyl-(1-->4)-D-xylopyranose (FAXX) were purified. The molecular masses of FAE-I and FAE-II were 69 and 24 kDa, respectively, under both denaturing and nondenaturing conditions. Apparent Km and maximum rate of hydrolysis with FAXX were 31.9 microM and 2.9 mumol min-1 mg-1 for FAE-I and 9.6 microM and 11.4 mumol min-1 mg-1 for FAE-II. FAE-II was specific for FAXX, but FAE-I hydrolyzed FAXX and PAXX, the equivalent p-coumaroyl ester, at a maximum rate of metabolism ratio of 3:1.
Borneman
W S
WS
Biotechnology Section, Merck & Co., Inc., Elkton, Virginia 22827.
Ljungdahl
L G
LG
Hartley
R D
RD
Akin
D E
DE
D002240:Carbohydrate Sequence; D002265:Carboxylic Ester Hydrolases / Q000302:isolation & purification* / Q000378:metabolism; D003373:Coumaric Acids / Q000378:metabolism*; D005658:Fungi / Q000201:enzymology*; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000378:metabolism
D003373:Coumaric Acids; D009844:Oligosaccharides; C004999:ferulic acid; D002265:Carboxylic Ester Hydrolases; C070196:feruloyl esterase