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Secretomic Analysis Reveals Multi-Enzymatic Complexes in Trichoderma reesei Grown in Media Containing Lactose or Galactose

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Abstract

The fungus Trichoderma reesei is a major producer of plant cell wall-degrading enzymes, such as cellulases and hemicellulases. These enzymes have been used to hydrolyze agro-industrial wastes in processes of second-generation biofuel production. The disaccharide lactose, the main by-product of dairy industry, is considered the most attractive substrate for the commercial production of cellulases by T. reesei. In the present work, it was shown that T. reesei, cultivated in media supplemented with either lactose or galactose, secrete proteins which behave as multi-enzymatic complexes. Blue Native electrophoresis (BN-PAGE) displayed two major bands which were specific or present in higher amounts in each secretome sample, complex I in galactose medium and complex II in lactose medium. The protein components of complexes I and II were identified by means of LC-MS/MS. The complexes were composed of different glycosyl hydrolases and predicted proteins related to biopolymer metabolism. Zymography coupled to BN-PAGE showed that T. reesei secretome presents higher cellulolytic and xylanolytic activities in lactose than in galactose-induced medium. The results presented here corroborate the existence of hydrolysis based on multi-enzymatic complexes in T. reesei secretome, as observed previously in other fungi, such as Trichoderma harzianum and Penicillium purpurogenum.

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Acknowledgments

This project was supported by the FAPDF (Federal District Foundation for Science and Research Supporting—grant nos. 193.000.584/2009 and 193.000.470/2011). FINEP (Design and Project Financing Agency), Petrobras (Brazilian Oil Corporation, CENPES 11007), CNPq (National Council for Scientific and Technological Development grant nos. 478066/2010-4 MCT/CNPq–Universal, 470265/2011-6, 563260/2010-6, and 563823/2010-0), University of Brasilia (UnB), Universidade Federal do Rio de Janeiro (UFRJ), and the National Institute of Science and Technology of Bioethanol. CNPq provided scholarship for DPGM.

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Authors and Affiliations

  1. Laboratório de Citogenética e Biologia Molecular, Departamento de Biologia e Zoologia, Universidade Federal de Mato Grosso, Cuiabá, 78060-900, MT, Brazil

    Adelson Joel da Silva

  2. Laboratório de Bioquímica e Química de Proteínas, Departamento de Biologia Celular, Universidade de Brasília, Brasília, 70910-900, DF, Brazil

    Diana Paola Gómez-Mendoza, Rayner Myr Lauterjung Queiroz, Marcelo Valle de Sousa & Carlos André Ornelas Ricart

  3. Laboratório de Enzimologia, Departamento de Biologia Celular, Universidade de Brasília, Brasília, 70910-900, DF, Brazil

    Edivaldo Ximenes Ferreira Filho

  4. Laboratório de Química de Proteínas – Unidade Proteômica, Instituto de Química, Universidade Federal do Rio de Janeiro, Rio de Janeiro, 21941-909, RJ, Brazil

    Magno Junqueira & Gilberto Barbosa Domont

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  1. Adelson Joel da Silva
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Correspondence to Carlos André Ornelas Ricart.

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da Silva, A.J., Gómez-Mendoza, D.P., Junqueira, M. et al. Secretomic Analysis Reveals Multi-Enzymatic Complexes in Trichoderma reesei Grown in Media Containing Lactose or Galactose. Bioenerg. Res. 8, 1906–1911 (2015). https://doi.org/10.1007/s12155-015-9648-4

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