between readings (Marx et al., 2001). A final fluorescence (Tf) was recorded by setting the fluorimeter to end point analysis at the end of the run. Various amounts of substrates were added to the samples to establish enzyme saturation (Chr6st, 1991). Incubations were carried out at 5, 10, 30, 60, 125, 250, and 500 pM for MUF-cellobioside; 5, 10, 20, 30, 50, 125, 250, and 500 [tM for MUF- phosphate; 5, 10, 20, 30, 50, 70, 100, 250, 500 pM for MUF-glucoside and 5, 10, 20, 50, 100, 300, 400, 800, and 1600 pM for L-Leucine amidomethylcoumarin. These substrate concentrations reflect in-well measurements as a consequence of dilution and are similar to the range of substrate concentrations used in another methodological study (Marx et al, 2001). In most cases fluorescence readings were taken over 2 hours. After evaluating the kinetic curves, the initial fluorescence (Ti) was subtracted from the final fluorescence (Tf) across incubation time to determine the potential enzyme activity. The largest difference in fluorescence (Tf-Ti) should yield the best resolution of activity for a given site. Alkalinization of the samples prior to fluorescence measurement was found unnecessary as all sample fluoresences were resolvable due to the high sensitivity of the spectrofluorimeter. Further conversion of potential enzyme activity to pmols MUF or AMC released g-1 AFDM h-1 was not necessary to meet the goals of this study. All figures represent averaged data with standard errors. Data from selected substrate concentrations are presented in figures to represent the extremes associated with each enzyme.