Cost-effective production of enzymes for industrial processes makes the appropriate selection of the host/vector expression system critical. We have tested two fungal systems for the bulk production of enzymes from thermophiles. The yeast Kluyveromyces lactis has been developed as a secretion host employing expression vectors based on the 2u-like plasmid pKD1 of Kluyveromyces drosophilarium. Our second system involves the filamentous fungus Trichoderma reesei. Signal and protein fusion vectors have been constructed using the strong cellobiohydrolase 1 (cbh1) promoter and recombinant plasmid DNAs introduced into various high-secreting T. reesei strains using biolistic particle delivery. In some cases (e.g. the xynB gene of Dictyoglomus thermophilum) we have reconstructed the genes according to Trichoderma codon preferences and demonstrated a dramatic increase in the production of the enzymes. The heterologous XynB enzyme is glycosylated differently in different Trichoderma strains. A proteomics approach has been taken to identify strongly expressed proteins produced by T. reesei under various cultivation conditions in order to identify condition-specific promoters driving the production of these proteins. Analyses indicated that HEX1, the major protein of the fungal Woronin body, is a dominant protein under both cellulase-inducing and -repressing conditions. The hex1 gene together with its promoter and terminator sequences has been isolated and the promoter function studied relative to cultivation time and medium.
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April 2004
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Conference Article|
April 01 2004
Recombinant enzymes from thermophilic micro-organisms expressed in fungal hosts
P.L. Bergquist;
P.L. Bergquist
1
*Department of Biological Sciences, and Research Institute for Biotechnology, Macquarie University, Sydney, New South Wales 2109, Australia
†Department of Molecular Medicine & Pathology, University of Auckland Medical School, Auckland, New Zealand
1To whom correspondence should be addressed, at the Research Institute for Biotechnology, Macquarie University (e-mail peter.bergquist@mq.edu.au).
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V.S.J. Te'o;
V.S.J. Te'o
*Department of Biological Sciences, and Research Institute for Biotechnology, Macquarie University, Sydney, New South Wales 2109, Australia
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M.D. Gibbs;
M.D. Gibbs
*Department of Biological Sciences, and Research Institute for Biotechnology, Macquarie University, Sydney, New South Wales 2109, Australia
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N.C. Curach;
N.C. Curach
*Department of Biological Sciences, and Research Institute for Biotechnology, Macquarie University, Sydney, New South Wales 2109, Australia
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K.M.H. Nevalainen
K.M.H. Nevalainen
*Department of Biological Sciences, and Research Institute for Biotechnology, Macquarie University, Sydney, New South Wales 2109, Australia
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Publisher: Portland Press Ltd
Online ISSN: 1470-8752
Print ISSN: 0300-5127
© 2004 Biochemical Society
2004
Biochem Soc Trans (2004) 32 (2): 293–297.
Citation
P.L. Bergquist, V.S.J. Te'o, M.D. Gibbs, N.C. Curach, K.M.H. Nevalainen; Recombinant enzymes from thermophilic micro-organisms expressed in fungal hosts. Biochem Soc Trans 1 April 2004; 32 (2): 293–297. doi: https://doi.org/10.1042/bst0320293
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