Figure 9.
Enzymatic hydrolysis of 4-methylumbelliferyl-ß-D-cellobioside (4MUC) with cellulase from T. reseei (Celluclast) applying a temperature profile in a MTP. (A) Progresses of 4-methyl-umbelliferyl (4MU) fluorescence (λex = 365 nm, λem = 455 nm) during the hydrolysis of 4MUC at different temperatures. Online data of 12 (from 48) exemplary wells. (B) Calibration curve for fluorescence intensity at varied 4MU concentration. (C) Temperature dependent initial reaction rate and (D) resulting Arrhenius plot of Celluclast. Dotted lines indicate Arrhenius fit. Experimental conditions: 96well MTP, VL = 200 μL, n = 995 rpm, d0 = 3 mm, 1 g/L Celluclast and 0.6 mM 4MUC in 0.1 M acetate buffer, pH = 4.8. Temperature profile: RT = 37°C, Tset,low = 10°C, Tset,high = 95°C (comp. Figure 3E).