Sophorose incubated

Figure 11. Polyacrylamide disc gel electrophoretic patterns of extracellular proteins produced by T. reesei QM 9414. The sample applied to the gel on the left was 130 fig extracellular protein from T. reesei my-celia grown on 1% Avicel (29), that applied to the gel on the right was 120 fig extracellular protein produced from sophorose-incubated mycelia. The bands shown here were stained for protein with Coomassie Blue and could, in all cases, also be stained for carbohydrate with the periodic acid-Schiff reagent.

Since essentially all of the protein in the cellulose-grown culture or the sophorose-incubation mixture can be accounted for by the three principal proteins and the / -glucosidase, it seems reasonable to conclude that the multiplicity of enzymes seen after several days of culture growth are caused by degradation. This may be attributable either to inherent instability (3) or to the presence of hydrolytic enzymes (16). There seems to be no evidence for the Ci enzyme as a nonhydrolytic or affinity factor, since all known enzymes of the system are hydrolytic. That the sophorose-stimulated system (with no possible loss of the Ci ... 

Figure 9. Enzyme production by T. reesei QM 9414 incubated in the presence of ImM sophorose. The incubation medium (27) included 17mM potassium phosphate buffer, pH 6.0, at 28°C. The appearance of aryl-p-D-glucosidase activity (A) in the extracellular medium is delayed in comparison to endoglucanase activity (O) and Avicelase activity ( ).

Figure 10. Product distribution from the incubation of enzymes (produced by T. reesei QM 9414 in response to sophorose) with phosphoric-acid swollen cellulose (PSC). Incubation time in this figure indicates the length of incubation of T. reesei mycelia in the presence of ImM sophorose. Enzymes produced at those times were incubated with PSC for 15 hr, and the products analyzed by HPLC (18/...

Pseudomonas fluorescens produced two extracellular (A and B) and one cell-bound (C) cellulase components, the latter being released by treatment with EDTA-lysozyme in isotonic sucrose. Culture with 0.5% glucose formed little cellulase. Cellobiose stimulated only the synthesis of C. The formation of A and B was strikingly enhanced in cultures with cellulose, sophorose, or continuous low concentration of cellobiose. The absence of extracellular cellulase synthesis in 0.5% cellobiose culture may be caused by catabolite repression. The three cellulases were purified and characterized. None of them split cellobiose, but all hydrolyzed various cellodextrins and celluloses. C easily attacked cellotriose and cellotriosyl sorbitol, but A and B had no effect. When pure B was incubated with broken spheroplasts of sophorose-grown cells, a cellulase component indistinguishable from A was formed. 

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