Preliminary investigation of fiber ultrastructure after enzymatic digestion
Knowledge on the chemical and physical structure of fiber is essential in determining forage quality. Current methods provide empirical results on fiber using chemical extractants. This paper is a preliminary report on methods to redefine fiber based on physiological (biochemical) procedures. TEM was employed to evaluate the relationship of fiber with specific enzymatic treatments.A modified AOAC Total Dietary Fiber Determination was used to prepare one group of insoluble fibers from two legumes of different nutritional quality. Three millimeter wedged-shaped leaflet sections of alfalfa (Medicago sativa L.) and 1-76 lespedeza [Sericea cuneata (Dumont)] were incubated stepwise with: (1) heat stable α amylase, pH 6.0, for h at 100°C; (2) protease, pH 7.5, for h at 60°C; and (3) amyloglucosidase, pH 4.5, for h at 60°C. All leaflet sections were retrieved and incubated for 24 h at 37°C with either: (1) 1% (w/v) cellulase in 0.05 M sodium acetate buffer, pH 5.0; (2) 10% (w/v) hemicellulase in 0.05 M acetate buffer, pH 5.5; or (3) 1% cellulase and 10% hemicel1ulase in the acetate buffer.