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Joosten, V. (author), Roelofs, M.S. (author), van den Dries, N. (author), Goosen, T. (author), Verrips, C.T. (author), van den Hondel, C.A.M.J.J. (author), Lokman, B.C. (author), TNO Kwaliteit van Leven (author)The Arthromyces ramosus peroxidase gene (arp) was genetically fused to either the 5′- or 3′-terminal ends of the gene encoding llama variable heavy chain antibody fragment VHH R9, resulting in the fusion expression cassettes ARP-R9 or R9-ARP. Aspergillus awamori transformants were obtained which produced up to 30 mg l-1 fusion protein in the...article 2005
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Michielse, C.B. (author), Hooykaas, P.J.J. (author), van den Hondel, C.A.M.J.J. (author), Ram, A.F.J. (author), TNO Kwaliteit van Leven (author)In the era of functional genomics, the need for tools to perform large-scale targeted and random mutagenesis is increasing. A potential tool is Agrobacterium-mediated fungal transformation. A. tumefaciens is able to transfer a part of its DNA (transferred DNA; T-DNA) to a wide variety of fungi and the number of fungi that can be transformed by...article 2005
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Joosten, V. (author), Gouka, R.J. (author), van den Hondel, C.A.M.J.J. (author), Verrips, C.T. (author), Lokman, B.C. (author), TNO Kwaliteit van Leven TNO Voeding (author)We report the expression and production of llama variable heavy-chain antibody fragments (VHHs) by Aspergillus awamori. Fragments encoding VHHs were cloned in a suitable Aspergillus expression vector and transformants secreting VHH fragments were analysed for integrated gene copy-numbers, mRNA levels and protein production. Functional V HHs were...article 2005
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Damveld, R.A. (author), van Kuyk, P.A. (author), Arentshorst, M. (author), Klis, F.M. (author), van den Hondel, C.A.M.J.J. (author), Ram, A.F.J. (author), TNO Kwaliteit van Leven TNO Voeding (author)1,3-α-d-Glucan is an important component of the cell wall of filamentous fungi. We have identified a family of five 1,3-α-d-glucan synthase-encoding genes in Aspergillus niger. The agsA gene was sequenced and the predicted protein sequence indicated that the overall domain structure of 1,3-α-d-glucan synthases is conserved in fungi. Using RT-PCR...article 2005
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Ram, A.F.J. (author), Arentshorst, M. (author), Damveld, R.A. (author), van Kuyk, P.A. (author), Klis, F.M. (author), van den Hondel, C.A.M.J.J. (author), TNO Voeding (author)Perturbation of cell wall synthesis in Saccharomyces cerevisiae, either by mutations in cell wall synthesis-related genes or by adding compounds that interfere with normal cell wall assembly, triggers a compensatory response to ensure cell wall integrity. This response includes an increase in chitin levels in the cell wall. Here it is shown that...article 2004
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Sigoillot, C. (author), Record, E. (author), Belle, V. (author), Robert, J.L. (author), Levasseur, A. (author), Punt, P.J. (author), van den Hondel, C.A.M.J.J. (author), Fournel, A. (author), Sigoillot, J.C. (author), Asther, M. (author), TNO Voeding (author)Three laccases, a natural form and two recombinant forms obtained from two different expression hosts, were characterized and compared for paper pulp bleaching. Laccase from Pycnoporus cinnabarinus, a well known lignolytic fungus, was selected as a reference for this study. The corresponding recombinant laccases were produced in Aspergillus...article 2004
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Michielse, C.B. (author), Ram, A.F.J. (author), van den Hondel, C.A.M.J.J. (author), TNO Voeding (author)The Aspergillus nidulans amdS selection marker was used for the identification of multicopy T-DNA insertions in Agrobacterium-mediated transformation of Asp. awamori. The selection of transformants on agar plates containing acetamide as sole nitrogen source and hygromycin resulted in a six-fold decrease in the transformation frequency, compared...article 2004
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Nelson, G. (author), Kozlova-Zwinderman, O. (author), Collis, A.J. (author), Knight, M.R. (author), Fincham, J.R.S. (author), Stanger, C.P. (author), Renwick, A. (author), Hessing, J.G.M. (author), Punt, P.J. (author), van den Hondel, C.A.M.J.J. (author), Read, N.D. (author), TNO Voeding Centraal Instituut voor Voedingsonderzoek TNO (author)Calcium signalling is little understood in filamentous fungi largely because easy and routine methods for calcium measurement in living hyphae have previously been unavailable. We have developed the recombinant aequorin method for this purpose. High levels of aequorin expression were obtained in Neurospora crassa, Aspergillus niger and...article 2004
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Al-Sheikh, H. (author), Watson, A.J. (author), Lacey, G.A. (author), Punt, P.J. (author), MacKenzie, D.A. (author), Jeenes, D.J. (author), Pakula, T. (author), Penttilä, M. (author), Alcocer, M.J.C. (author), Archer, D.B. (author), TNO Voeding (author)We describe a new endoplasmic reticulum (ER)-associated stress response in the filamentous fungus Aspergillus niger. The inhibition of protein folding within the ER leads to cellular responses known collectively as the unfolded protein response (UPR) and we show that the selective transcriptional downregulation of the gene encoding glucoamylase,...article 2004
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TNO Voeding Centraal Instituut voor Voedingsonderzoek TNO (author), Levasseur, A. (author), Pagès, S. (author), Fierobe, H.P. (author), Navarro, D. (author), Punt, P. (author), Belaïch, J.P. (author), Asther, M. (author), Record, E. (author)A chimeric enzyme associating feruloyl esterase A (FAEA) from Aspergilhis niger and dockerin from Clostridium thermocellum was produced in A. niger. A completely truncated form was produced when the dockerin domain was located downstream of the FAEA (FAEA-Doc), whereas no chimeric protein was produced when the bacterial dockerin domain was...article 2004
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TNO Bouw (author), Sailer, M.F. (author), van Etten, B.D. (author)Due to the increasing restrictions in the use of wood preserving biocides a number of potential biocide free wood preserving alternatives are currently assessed. Wood degrading fungi require certain conditions in the wood in order to be able to use wood as a food source. This paper discusses the physiological requirements of wood degrading fungi...article 2004
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Joosten, V. (author), Lokman, C. (author), van den Hondel, C.A.M.J.J. (author), Punt, P.J. (author), TNO Voeding (author)In this review we will focus on the current status and views concerning the production of antibody fragments and antibody fusion proteins by yeasts and filamentous fungi. We will focus on single-chain antibody fragment production (scFv and VHH) by these lower eukaryotes and the possible applications of these proteins. Also the coupling of...article 2003
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Record, E. (author), Asther, M. (author), Sigoillot, C. (author), Pagès, S. (author), Punt, P.J. (author), Delattre, M. (author), Haon, M. (author), van den Hondel, C.A.M.J.J. (author), Sigoillot, J.C. (author), Lesage-Meessen, L. (author), Asther, M. (author), TNO Voeding (author)A well-known industrial fungus for enzyme production, Aspergillus niger, was selected to produce the feruloyl esterase FAEA by homologous overexpression for pulp bleaching application. The gpd gene promoter was used to drive FAEA expression. Changing the nature and concentration of the carbon source nature (maltose to glucose; from 2.5 to 60 g l...article 2003
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Punt, P.J. (author), Drint-Kuijvenhoven, A. (author), Lokman, B.C. (author), Spencer, J.A. (author), Jeenes, D. (author), Archer, D.A. (author), van den Hondel, C.A.M.J.J. (author)We have characterized growth and protein processing characteristics of Aspergillus niger strains carrying a disrupted allele of the previously cloned and characterized kexB gene [Appl. Environ. Microbiol. 66 (2000) 363] encoding a furin-type endoprotease. Deletion of the single-copy gene confirms it to be non-essential but disruptant strains...article 2003
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Lokman, B.C. (author), Joosten, V. (author), Hovenkamp, J. (author), Gouka, R.J. (author), Verrips, C.T. (author), van den Hondel, C.A.M.J.J. (author)The heterologous production of Arthromyces ramosus peroxidase (ARP) was analysed in the filamentous fungus Aspergillus awamori under control of the inducible endoxylanase promoter. Secretion of active ARP was achieved up to 800 mg l-1 in shake flask cultures. Western blot analysis showed that an rARP product of the correct molecular weight was...article 2003
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TNO Voeding Centraal Instituut voor Voedingsonderzoek TNO (author), Yi, X. (author), Conesa, A. (author), Punt, P.J. (author), Hager, L.P. (author)Site-directed mutagenesis has been used to investigate the role of glutamic acid 183 in chloroperoxidase catalysis. Based on the x-ray crystallographic structure of chloroperoxidase, Glu-183 is postulated to function on distal side of the heme prosthetic group as an acid-base catalyst in facilitating the reaction between the peroxidase and...article 2003
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te Biesebeke, R. (author), Ruijter, G. (author), Rahardjo, Y.S.P. (author), Hoogschagen, M.J. (author), Heerikhuisen, M. (author), Levin, A. (author), van Driel, K.G.A. (author), Schutyser, M.A.I. (author), Dijksterhuis, J. (author), Zhu, Y. (author), Weber, F.J. (author), de Vos, W.M. (author), van den Hondel, K.A.M.J.J. (author), Rinzema, A. (author), Punt, P.J. (author)We report the progress of a multi-disciplinary research project on solid-state fermentation (SSF) of the filamentous fungus Aspergillus oryzae. The molecular and physiological aspects of the fungus in submerged fermentation (SmF) and SSF are compared and we observe a number of differences correlated with the different growth conditions. First,...article 2002
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Wolken, W.A.M. (author), van Loo, W.J.V. (author), Tramper, J. (author), van der Werf, M.J. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)A novel lyase, combining hydratase and aldolase activity, that converts citral into methylheptenone and acetaldehyde, was purified from spores of Penicillium digitatum. Remarkably, citral lyase activity was induced 118-fold by incubating nongerminating spores with the substrate, citral. This cofactor independent hydratase/aldolase, was purified...article 2002
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Punt, P.J. (author), van Biezen, N. (author), Conesa, A. (author), Albers, A. (author), Mangnus, J. (author), van den Hondel, C. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)Filamentous fungi have been used as sources of metabolites and enzymes for centuries. For about two decades, molecular genetic tools have enabled us to use these organisms to express extra copies of both endogenous and exogenous genes. This review of current practice reveals that molecular tools have enabled several new developments. But it has...article 2002
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Conesa, A. (author), Punt, P.J. (author), Hondel, C.A.M.J.J. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)Peroxidases are oxidoreductases that utilize hydrogen peroxide to catalyze oxidative reactions. A large number of peroxidases have been identified in fungal species and are being characterized at the molecular level. In this manuscript we review the current knowledge on the molecular aspects of this type of enzymes. We present an overview of the...article 2002