Cellulase components induced by sophorose and cell...
Cellulose and sophorose supplementation of the glycerol medium induced the extracellular and intracellular cellulase components Cxand cellobiase（see Figure 3）.C1 cellulase induction was also shown in another experiment.
The spore suspension of EA3-768 was inoculated on glycerol medium with and without cellulose（0.5%，added at 0 h）or sophorose（10-4 M，added after two days in culture）supplementation，cultured at 33℃to reach maximum cellulase production.The extracellular and intracellular cellulases were extracted from the culture filtrates and mycelia，respectively，precipitated by acetone at-20℃，dissolved in acetate buffer（0.1 M，pH=4.8），and separated by electrophoresis on polyacrylamide disc gel（see Figure 4）.All gels showed the very similar pattern of proteins（Zones Ⅰ—Ⅵ）.According to the assay of cellulase activity of each gel band，bands Ⅲ，Ⅳ and Ⅴ were identified as C1（cotton ceUulase），Cx（CM cellulase）andβ-glucosidase，respectively.Clearly，C1，Cxandβglucosidase were strongly induced by cellulose，and especially by sophorose.In the glycerol control，a very small amount of C1，Cxand β-glucosidase，presumed to be constitutive cellulase，was detected.It is of interest that the level of intracellular cellobiase in the glycerol control was distinctly higher than the extracellular one.
Figure 3 Comparison of distribution of extracellular（a，c）and intracellular（b，d）cellulase Cx，cellobiase of EA3-867 induced by sophorose and cellulose added to glycerol medium.●，10-4 Ms sephorose（SP）added after 2 days；o，0.5%cellulose powder（CP）added at zero time［5］
Figure 4 Schematic comparison of polyaceylamide gel electrophoresis patterns of Trichoderma EA3-867 induced by sophorose（SR）.cellulose powder（CP）and no inducer（CK）.［5］Electrophoresis conditions：cellulase 400 μg.current 2mA/tube，time 1.5 h.Ⅲ.Cotton cellulase；Ⅳ.CM cellulase；Ⅴ.β-glucosidase
