Separation，purification and properties of cellulas...
Since sophorose coordinately induces cellulase C1，Cxandβ-glucosidase of washed mycelium of Trichoderma to high levels（see Figure 8），this preparation was used as starting material for purification.
After DEAE-Sephadex A-50 chromatography C1，Cx，andβ-glucosidase were separated.The C1 protein was further purified by preparative gel electrophoresis.The Cx component andβ-glucosidase were separated by passing through Sephadex G-100.Selective inactivation was also used.［9］(https://www.daowen.com)
Figure 8 Changes in extracellular cellulase，soluble protein，and RNA of mycelium during induction of washed mycelium of N2-78 by sophorose.2 Mycelium：1.5 mg/ml；sophorose：10-4 M.C1is expressed by cotton cellulase，Cx（R）and Cx（V）represent CM cellulase measured by reducing sugar increase or by viscosity decrease，respectively.○，Cx（V）；●，Cx（R）；■，C1；▲，βglucosidase；×，extracellular soluble protein；△，RNA
The molecular weights of C1，Cxandβ-glucosidase were estimated to be 67，000，62，000 and 42，000，respectively，on polyacrylamide gel［9］The C1 component was shown to be homogenous by polyacrylamide gel and cellulose acetate film electrophoresis，immunoelectrophoresis and ultracentrifugal analysis（see Figure 9）.Component C1 was a glycoprotein containing 16 amino acids（mainly glycine，aspartic acid，threonine，serine and glutamic acid）.The C1 component showed a strong synergistic action with Cxcomponent in the degradation of cotton，Avicel and Walseth cellulose.The hydrolysates of Walseth cellulose by C1 action were mainly cellobiose and a trace of glucose.This supports the concept that C1action is mainly as a cellobiohydrolase.The stability of these components to high temperature was lowered when they were separated.
Figure 9 Identification of C1-cellulase by polyacrylamide gel electrophoresis，cellulase acetate film electrophoresis，immuno electrophoresis and ultracentrifugal analysis［9］
