New Effective Method for Analysis of the Component Composition of Enzyme Complexes from Trichoderma reesei

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A. V. Markov^1*, A. V. Gusakov¹, E. G. Kondratyeva¹, O. N. Okunev², A. O. Bekkarevich², and A. P. Sinitsyn¹

¹Faculty of Chemistry, Lomonosov Moscow State University, 119992 Moscow, Russia; fax: (7-095) 939-0997; E-mail: avmarkov@enzyme.chem.msu.ru

²Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, 142292 Pushchino, Moscow Region, Russia; fax: (7-095) 923-3602; E-mail: oleg@ibpm.serpukhov.su

^* To whom correspondence should be addressed.

Received May 21, 2004; Revision received June 25, 2004

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A method for analysis of the component composition of multienzyme complexes secreted by the filamentous fungus Trichoderma reesei was developed. The method is based on chromatofocusing followed by further identification of protein fractions according to their substrate specificity and molecular characteristics of the proteins. The method allows identifying practically all known cellulases and hemicellulases of T. reesei: endoglucanase I (EG I), EG II, EG III, cellobiohydrolase I (CBH I), CBH II, xylanase I (XYL I), XYL II, beta-xylosidase, alpha-L-arabinofuranosidase, acetyl xylan esterase, mannanase, alpha-galactosidase, xyloglucanase, polygalacturonase, and exo-beta-1,3-glucosidase. The component composition of several laboratory and commercial T. reesei preparations was studied and the content of the individual enzymes in these preparations was quantified. The influence of fermentation conditions on the component composition of secreted enzyme complexes was revealed. The characteristic features of enzyme preparations obtained in “cellulase” and “xylanase” fermentation conditions are shown.

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KEY WORDS: Trichoderma reesei, chromatofocusing, component composition, cellulases, hemicellulases, fermentation

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