We assessed the e †ect of ferulate substitution and diferulate crosslinking of xylans on the degradation of cell walls by two fungal enzyme mixtures, one of which contained feruloyl esterase and high xylanase activities. Non-ligniÐed cell suspensions of maize (Zea mays) were grown with 0 or 40 lM 2aminoindan-2-phosphonic acid to produce walls with normal (17É2 mg g~1) or reduced (5É1 mg g~1) ferulate concentrations. Walls were incubated with mercaptoethanol to inhibit diferulate formation or with hydrogen peroxide to stimulate diferulate formation by wall bound peroxidases. Varying the ferulate substitution of xylans did not a †ect cell wall hydrolysis. In contrast, increasing ferulate dimerisation from 18 to 40% reduced carbohydrate release by 94È122 mg g~1 after 3 h and by 0È48 mg g~1 after 54 h of enzymatic hydrolysis. Diferulate crosslinks impeded the release of xylans, cellulose and pectins from walls. These results provide compelling evidence that diferulate cross-links reduce the rate and, to a lesser degree, the extent of wall hydrolysis by fungal enzymes. Our results also suggest that enzyme mixtures containing high xylanase activity but not feruloyl esterase activity can partially overcome the inhibitory e †ects of diferulate cross-linking on wall hydrolysis.
1998 SCI.