Proteome and transcriptome analysis of the cellular responses to production of the heterologous protein, Melanocarpus albomyces laccase, in the filamentous fungus Trichoderma reesei

Tiina Pakula (Corresponding author), Jari Rautio, Bart Smit, Mikko Arvas, Markku Saloheimo, Merja Penttilä

    Research output: Chapter in Book/Report/Conference proceedingConference abstract in proceedingsScientific


    Trichoderma reesei Rut-C30 and its transformant producing Melanocarpus
    laccase were cultivated in lactose-containing minimal medium in
    chemostats and subjected to proteome and transcriptome analysis to identify
    differentially expressed genes under these conditions. The cultures were carried
    out using a dilution rate (0.03/h) previously shown to be optimal for production
    of endogenous extracellular proteins under these conditions. The TRAC method
    for multiplexed transcriptional analysis was first applied to analyse the
    expression levels of a selected set of genes and to monitor the steady state in
    the chemostat cultures. The results indicated that UPR (unfolded protein
    response) pathway was not activated as a response to laccase production. On
    the contrary, the expression levels of e.g. the foldase and chaperone genes pdi1
    and bip1 were expressed at a slightly higher level in the host. Also the
    expression levels of the genes encoding the endogenous secreted cellulases,
    egl1 and cbh1, were slightly higher in the host strain. Both proteome and
    genome wide transcriptional analysis using oligonucleotide microarrays revealed
    only a modest number of cellular responses to laccase production. Altogether 32
    genes were differentially expressed between the strains, only five of the genes
    being upregulated in the laccase producing strain, and the rest having a higher
    expression level in the host strain and including many genes encoding
    extracellular enzymes. Proteome analysis using 2D gel electrophoresis with DIGE
    showed increased expression of a group of proteins including e.g. heat shock
    proteins as well ubiquitin associated proteins and proteins involved in ERassociated protein degradation. The result indicates that although laccase
    expression does not induce apparent UPR, other type stress responses are
    induced e.g. activated protein degradation. The results were also compared to
    those obtained in previous studies on the production human tissue plasminogen
    activator (tPA) in T. reesei.
    Original languageEnglish
    Title of host publication8th European Conference on Fungal Genetics
    Subtitle of host publicationBook of Abstracts
    EditorsIrina S. Druzhnina, Alexey G. Kopchinskiy, Christian P. Kubicek
    PublisherVienna University of Technology
    ChapterVII Cell-Factories and Biotechnology
    Publication statusPublished - 2006
    Event8th European Conference on Fungal Genetics - Vienna, Austria
    Duration: 8 Apr 200611 Apr 2006


    Conference8th European Conference on Fungal Genetics


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