Abstract
Cell walls of Coastal Bermuda grass ( Cynodon dactylon) were treated with polysaccharide hydrolases to release O-[5- O-( trans-p-coumaroyl)-α- l-arabinofuranosyl]-(1→3)- O-β- d-xylopyranosyl-(1→4)- d-xylopyranose (PAXX) which was isolated by liquid chromatography. The isolated PAXX was greater than 95% pure as determined by 1H NMR and was used as substrate for a sensitive assay of trans-p-coumaroyl esterase. PAXX was hydrolyzed by culture filtrates from the anaerobic fungus Neocallimastix MC-2. The trans-p-coumaric acid released by enzymatic hydrolysis was assayed by reverse-phase HPLC, and as little as 100 ng of acid could be determined. Steady-state velocities for the release of the acid obeyed Michaelis-Menten kinetics. V max was determined to be 1.17 μmol min −1 mg −1 and K m 13.2 μ m at pH 7.5 and 30°C.
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