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Hydrolysis of Cellulose by Saturating and Non–Saturating Concentrations of Cellulase: Implications for Synergism

Abstract

The adsorption of purified cellobiohydrolases (CBH I and II) and endoglucanases (EG I and II) from Trichoderma reesei strain L27 to microcrystalline cellulose (Avicel) has been studied. Scatchard analysis of the adsorption data indicated that Avicel possessed high– and low–affinity binding sites for the cellulase components and gave the maximum amount of each component that bound to Avicel at saturation. Hydrolysis of Avicel was thus carried out by saturating and non–saturating concentrations of purified cellulase components alone and in combination with each other. Synergism between them was greatest when Avicel was incubated with non–saturating concentrations of EG I or EG II with CBH I and CBH II. This finding could lead to a dramatic reduction in the enzyme requirement for cellulose utilization.

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References

  1. Shoemaker, S., Watt, K., Tsitovsky, G. and Cox, R. 1983. Characterization and properties of cellulases purified from Trichoderma reesei strain L27. Bio/Technology 1:687–690.

    CAS  Google Scholar 

  2. Fägerstam, L.G. and Pettersson, L.G. 1980. The 1,4-β-glucan cello-biohydrolases of Trichoderma reesei QM 9414. A new type of cellulolytic synergism. FEES Lett. 119:97–100.

    Article  Google Scholar 

  3. Beldman, G., Searle-Van Leewven, M.F., Rombouts, F.M. and Voragen, F.G.J. 1985. The cellulase of Trichoderma viride. Eur. J. Biochem. 146:301–308.

    CAS  Article  Google Scholar 

  4. Chernoglazov, V.M., Ermolova, O.V. and Klësov, A.A. 1985. Production of highly purified multiple forms of endo-1,4-β-glucanases from Trichoderma viride, differing in adsorption capacity by affinity chromatography on cellulose and highly effective chromatofocusing. Biokhimiya 50:1108–1119.

    CAS  Google Scholar 

  5. Wood, T.M. 1985. Properties of cellulolytic enzyme systems. Biochem. Soc. Trans. 13:407–410.

    CAS  Article  Google Scholar 

  6. Wood, T.M. and McCrae, S.I. 1982. Purification and some properties of the extracellular (β-D-glucosidase of the cellulolytic fungus Trichoderma koningii. J. Gen. Microbiol. 128:2973–2982.

    CAS  Google Scholar 

  7. Wood, T.M. and McCrae, S.I. 1986. The cellulase of Penicillium pinophilium. Biochem. J. 234:93–99.

    CAS  Article  Google Scholar 

  8. Stuart, J.Y. and Ristroph, D.L. 1985. Analysis of cellulosc-cellulase adsorption data: a fundamental approach. Biotechnol. Bioeng. 27:1056–1059.

    CAS  Article  Google Scholar 

  9. Converse, A.O. and Grethlein, H.E. 1987. On the use of an adsorption model to represent the effect of steam explosion pretreatment on the enzymatic hydrolysis of lignocellulosic substances. Enzyme Microb. Technol. 9:79–82.

    CAS  Article  Google Scholar 

  10. Peitersen, N., Medeiros, J. and Mandels, M. 1977. Adsorption of Trichoderma cellulase on cellulose. Biotechnol. Bioeng. 19:1091–1094.

    CAS  Article  Google Scholar 

  11. Ryu, D.D.Y., Kim, C. and Mandels, M. 1984. Competitive adsorption of cellulose components and its significance in a synergistic mechanism. Biotechnol. Bioeng. 26:488–496.

    CAS  Article  Google Scholar 

  12. Reese, E.T. 1982. Elution of cellulase from cellulose. Process Biochem. 17:2–8.

    CAS  Google Scholar 

  13. Grethlein, H.E. 1985. The effect of pore size distribution on the rate of enzymatic hydrolysis of cellulosic substrates. Bio/Technology 3:155–160.

    CAS  Article  Google Scholar 

  14. Klysov, A.A., Mitkevich, O.V. and Sinitsyn, A.P. 1986. Role of the activity and adsorption of cellulases in the efficiency of the enzymatic hydrolysis of amorphous and crystalline cellulose. Biochemistry 25:540–542.

    Article  Google Scholar 

  15. Henrissat, B., Driguez, H., Viet, C. and Shuülein, M. 1985. Synergism of cellulases from Trichoderma reesei in the degradation of cellulose. Bio/Technology 3:722–726.

    CAS  Article  Google Scholar 

  16. Rabinovich, M.L., Van Viet, N. and Klesov, A.A. 1982. Adsorption of cellulolytic enzymes on cellulose and kinetics of the action of adsorbed enzymes. Two types of interaction of the enzymes with an insoluble substrate. Biokhimiya 47:465–477.

    CAS  Google Scholar 

  17. Chanzy, H., Henrissat, B. and Vuong, R. 1984. Colloidal gold labelling of 1,4-β-D-glucan cellobiohydrolase adsorbed on cellulose substrates. FEES Lett. 172:193–197.

    CAS  Article  Google Scholar 

  18. Woodward, J., Marquess, H.J. and Picker, C.S. 1986. Affinity chromatography of β-glucosidase and endo-β-glucanase from Aspergillus niger on concanavalin A-Sepharose: Implications for cellulase component purification and immobilization. Prep. Biochem. 16:337–352.

    CAS  PubMed  Google Scholar 

  19. Segel, I.H. 1975. Enzyme Kinetics. John Wiley, New York.

    Google Scholar 

  20. Woodward, J. and Arnold, S.A. 1981. The inhibition of β-glucosidase activity in Trichoderma reesei C30 cellulase by derivatives and isomers of glucose. Biotechnol. Bioeng. 23:1553–1562.

    CAS  Article  Google Scholar 

  21. Miller, G.L. 1959. Use of denitrosalicyclic acid reagent for determination of reducing sugar. Anal. Chem. 31:426–428.

    CAS  Article  Google Scholar 

  22. Bradford, M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72:248–254.

    CAS  Article  Google Scholar 

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Woodward, J., Hayes, M. & Lee, N. Hydrolysis of Cellulose by Saturating and Non–Saturating Concentrations of Cellulase: Implications for Synergism. Nat Biotechnol 6, 301–304 (1988). https://doi.org/10.1038/nbt0388-301

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