Trichoderma reesei cellobiohydrolases from

Figure 5 shows the results of inhibition experiments involving the D-gluco-sides 37 (n = 1-3) and a cellobiohydrolase from Trichoderma reesei. The best inhibition was achieved with the epoxypentyl /J-D-C-glucoside, a result which corresponds nicely to the 0-glycoside series where the epoxybutyl -o-gluco-side 6 (n = 2) was the most effective inhibitor. 

Compound (44 g, NHAc form) (Scheme 14) was found to be a competitive inhibitor for CBHI cellulase (family 7) from Trichoderma reesei, when 4-methyl-umbelliferyl / -lactoside was used as substrate. Therefore (44 g, NHj form) was coupled to CH-Sepharose 4B, and the affinity gel was very effective for the purification of cellobiohydrolases from a crude commercial cellulolytic extract of T. reesei [40c]. Using the same approach aryl 1,4-dithioxylobioside and l,4,4 -trithioxylotrioside (44 h, NH2 form) were coupled to CH-Sepharose 4B to give affinity gels which were used for the purification of xylanases [40a,b]. 

Divne, C., Stahlberg.J., Reinikainen, T., Ruohonen, L., Pettersson, G., KnowlesJ., et al. (1994) The three-dimensional crystal stractme of the catalytic core of cellobiohydrolase I from Trichoderma reesei. Science, 265, 524-528. 

Divne, C., Stahlberg, J., Teeri, T. and Jones, T. (1998) High-resolution crystal structures reveal how a cellulose chain is bound in the 50 Angstrom long tuimel of cellobiohydrolase I from Trichoderma reesei. J. Mol. Biol., 275, 309-325. 

Medve, J., Karlsson, J., Lee, D., and Tjerneld, F. 1998. Hydrolysis of microcrystalline cellulose by cellobiohydrolase I and endoglucanase II from Trichoderma reesei Adsorption, sugar production pattern and synergism of the enzymes. Biotech. Bioeng.,59, 621-634. 

Reinikainen, T. 1994. The Cellulose-Binding Domain of Cellobiohydrolase I from Trichoderma reesei (pg. 206). Espoo, Finland VTT publications. 

Reinikainen, T., Teleman, O., and Teeri, T. T. 1995. Effects of Ph and High Ionic-Strength on the Adsorption and Activity of Native and Mutated Cellobiohydrolase-I from Trichoderma-Reesei. Proteins-Structure Function and Genetics, 22, 392-403. 

Vrsanska, M. and Biely, P., The cellobiohydrolase I from Trichoderma reesei QM 9414 Action on cello-oligosaccharides. Carbohydrate Res 1992, 227, 19 27. 

Lemos, M. A., Teixeira, J. A., Domingues, M. R. M., Mota, M., and Gama, F. M., The enhancement of the cellulolytic activity of cellobiohydrolase I and endoglucanase by the addition of cellulose binding domains derived from Trichoderma reesei. Enzyme Microbial Technol 2003, 32, (1), 35—40. 

Reinikainen T, Teleman O, Teeri TA (1995) Effects of pH and high ionic strength on the adsorption and activity of native and mutated cellobiohydrolase 1 from Trichoderma reesei. Proteins 22 392- 403... 

Shoemaker S, Schweickart V, Ladner M, Gelfand D, Kwok S, Myambo K, Innis M (1983) Molecular cloning of exo-cellobiohydrolase I derived from Trichoderma reesei strain L27. Bio/Technology 1 691-699... 

Medve J, Stahlberg J, Tjernel F (1994) Adsorption and synergism of cellobiohydrolase I and II from Trichoderma reesei during hydrolysis of microcrystalline cellulose. Biotechnology and Bioengineering 44 1064 -1073... 

Abuja PM, Schmuck M, Pilz 1, Tomme P, Claeyssens M, Esterbauer H (1988) Structural and functional domains of cellobiohydrolase I from Trichoderma reesei. European Biophysics Journal 15 339-342... 

Van Tilbeurgh H, Tomme P, Claeyssens M, Bikhabai R, Pettersson G (1986) Limited proteolysis of the cellobiohydrolase I from Trichoderma reesei-. separation of the functional domains. FEBS Letters 204 223-227... 

Rouvinen J, Bergfors T, Teeri T, Knowles JKC, Jones TA (1990) Three-dimensional structure of cellobiohydrolase II from Trichoderma reesei. Science 249 380-385 Srisodsuk M, Reinikainen T, PenttUa M, Teeri TT (1993) Role of the interdomain linker peptide of Trichoderma reesei cellobiohydrolase I in its interaction with crystalline cellulose. Journal of Biological Chemistry 268 20,756-20,761... 

Figure 6. Fractionation of tiyptic peptides of carboxymethylated 1,4-glucan cellobiohydrolase fromll Trichoderma reesei on Mono Q, monitored at 214 nm. Sanriple, 80 min tryptic digest flow-rate, 1 ml/min eluant, 20 mM Tris—HCI. pH 8.0 with NaCl gradient 2.5 mM—0.5 M. (From Pharmacia Fine Chemicals AB, Uppsala, Sweden.) i...

Woodward, J. Tate, J. Herrmann, P. C. Evans, B. R. Comparison of Ellman s reagent with N-(l-pyrenyl)maleimide for the determination of free sulfhydryl groups in reduced cellobiohydrolase I from Trichoderma reesei. J. Biochem. Biophys. Methods 1993,26,121-129. 

Cellulase-Negative Xylanase-Positive Mutants. There are two reports concerning the selection of such mutants from filamentous fungi, one on Pofyporus adm-tus (32) and the other cmi Trichoderma reesei (Durand, H. et al. Society CAYLA, Toulouse, France, unpublished results). An analysis of the eliminated cellulase genes has not been done, so it is not known if the mutants negative in endo(l- 4)-p-glucan-ase were deficient also in cellobiohydrolases. 

Figure 1. Fractionation of proteins in the culture filtrate of Trichoderma reesei according to their pi values Xyl, xylanase Ara, arabinosidase AE, acetyl esterase / X, /3-xylosidase aG, a-glucuronidase / G, / -glucosidase CBH, cellobiohydrolase EG, endoglucanase. Chromatofocusing was performed in a PBE-94 anion exchange resin (Pharmacia) with a pH-gradient created by ampholyte buffers (Pharmacia). Solid line, A dotted line, pH. (Reproduced with permission from ref. 24. Copyright 1988.)...

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