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The intracellular galactoglycome in Trichoderma reesei during growth on lactose

  • Applied genetics and molecular biotechnology
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Abstract

Lactose (1,4-0-β-d-galactopyranosyl-d-glucose) is used as a soluble carbon source for the production of cellulases and hemicellulases for—among other purposes—use in biofuel and biorefinery industries. The mechanism how lactose induces cellulase formation in T. reesei is enigmatic, however. Previous results from our laboratory raised the hypothesis that intermediates from the two galactose catabolic pathway may give rise to the accumulation of intracellular oligogalactosides that could act as inducer. Here we have therefore used high-performance anion-exchange chromatography–mass spectrometry to study the intracellular galactoglycome of T. reesei during growth on lactose, in T. reesei mutants impaired in galactose catabolism, and in strains with different cellulase productivities. Lactose, allo-lactose, and lactulose were detected in the highest amounts in all strains, and two trisaccharides (Gal-β-1,6-Gal-β-1,4-Glc/Fru and Gal-β-1,4-Gal-β-1,4-Glc/Fru) also accumulated to significant levels. Glucose and galactose, as well as four further oligosaccharides (Gal-β-1,3/1,4/1,6-Gal; Gal-β-1,2-Glc) were only detected in minor amounts. In addition, one unknown disaccharide (Hex-β-1,1-Hex) and four trisaccharides were also detected. The accumulation of the unknown hexose disaccharide was shown to correlate with cellulase formation in the improved mutant strains as well as the galactose pathway mutants, and Gal-β-1,4-Gal-β-1,4-Glc/Fru and two other unknown hexose trisaccharides correlated with cellulase production only in the pathway mutants, suggesting that these compounds could be involved in cellulase induction by lactose. The nature of these oligosaccharides, however, suggests their formation by transglycosylation rather than by glycosyltransferases. Based on our results, the obligate nature of both galactose catabolic pathways for this induction must have another biochemical basis than providing substrates for inducer formation.

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Acknowledgments

This work was supported by grants from the Austrian Science Foundation (FWF P-19143 and FWF P24219) to CPK and BS, respectively. Research at the University of Debrecen has been supported by the OTKA (Hungarian Scientific Research Fund; grants K67667 and K1006600) and by the TÁMOP-4.2.2/B-10/-1-2010-0024 Project. EF is a recipient of a Bólyai János Research Scholarship (BO/00519/09/8).

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Authors and Affiliations

  1. Department of Biochemical Engineering, Faculty of Science and Technology, University of Debrecen, 4032, Egyetem tér 1, Debrecen, Hungary

    Levente Karaffa & Erzsébet Fekete

  2. TNO, P.O. Box 360, 3700 AJ, Zeist, the Netherlands

    Leon Coulier, Karin M. Overkamp & Peter J. Punt

  3. Research Area Biotechnology and Microbiology, Institute of Chemical Engineering, TU Wien, Gumpendorferstrasse 1a, 1060, Vienna, Austria

    Irina S. Druzhinina, Marianna Mikus, Bernhard Seiboth & Christian P. Kubicek

  4. Austrian Center of Industrial Biotechnology (ACIB), c/o TU Wien, Gumpendorferstrasse 1a, 1060, Vienna, Austria

    Irina S. Druzhinina, Marianna Mikus, Bernhard Seiboth & Christian P. Kubicek

  5. Department of Colloid and Environmental Chemistry, Faculty of Science and Technology, University of Debrecen, 4032, Egyetem tér 1, Debrecen, Hungary

    Levente Novák

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  1. Levente Karaffa
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  2. Leon Coulier
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  10. Christian P. Kubicek
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Correspondence to Levente Karaffa.

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Karaffa, L., Coulier, L., Fekete, E. et al. The intracellular galactoglycome in Trichoderma reesei during growth on lactose. Appl Microbiol Biotechnol 97, 5447–5456 (2013). https://doi.org/10.1007/s00253-012-4667-y

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  • DOI: https://doi.org/10.1007/s00253-012-4667-y

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