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Karnaukhova, E. (author), Ophir, Y. (author), Trinh, L. (author), Dalal, N. (author), Punt, P.J. (author), Golding, B. (author), Shiloach, J. (author), TNO Kwaliteit van Leven (author)Background: Human α1-proteinase inhibitor (α1-PI), also known as antitrypsin, is the most abundant serine protease inhibitor (serpin) in plasma. Its deficiency is associated with development of progressive, ultimately fatal emphysema. Currently in the United States, α1-PI is available for replacement therapy as an FDA licensed plasma-derived (pd...article 2007
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Goosen, C. (author), Yuan, X.L. (author), van Munster, J.M. (author), Ram, A.F.J. (author), van der Maarel, M.J.E.C. (author), Dijkhuizen, L. (author), TNO Kwaliteit van Leven (author)A novel subfamily of putative intracellular invertase enzymes (glycoside hydrolase family 32) has previously been identified in fungal genomes. Here, we report phylogenetic, molecular, and biochemical characteristics of SucB, one of two novel intracellular invertases identified in Aspergillus niger. The sucB gene was expressed in Escherichia...article 2007
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van der Kaaij, R.M. (author), Yuan, X.L. (author), Franken, A. (author), Ram, A.F.J. (author), Punt, P.J. (author), van der Maarel, M.J.E.C. (author), Dijkhuizen, L. (author), TNO Kwaliteit van Leven (author)In the genome sequence of Aspergillus niger CBS 513.88, three genes were identified with high similarity to fungal α-amylases. The protein sequences derived from these genes were different in two ways from all described fungal α-amylases: they were predicted to be glycosylphosphatidylinositol anchored, and some highly conserved amino acids of...article 2007
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van der Kaaij, R.M. (author), Janeček, Š. (author), van der Maarel, M.J.E.C. (author), Dijkhuizen, L. (author), TNO Kwaliteit van Leven (author)Currently known fungal α-amylases are well-characterized extracellular enzymes that are classified into glycoside hydrolase subfamily GH13_1. This study describes the identification, and phylogenetic and biochemical analysis of novel intracellular fungal α-amylases. The phylogenetic analysis shows that they cluster in the recently identified...article 2007
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Yuan, X.L. (author), Goosen, C. (author), Kools, H. (author), van der Maarel, M.J.E.C. (author), van den Hondel, C.A.M.J.J. (author), Dijkhuizen, L. (author), Ram, A.F.J. (author), TNO Kwaliteit van Leven (author)As a soil fungus, Aspergillus niger can metabolize a wide variety of carbon sources, employing sets of enzymes able to degrade plant-derived polysaccharides. In this study the genome sequence of A. niger strain CBS 513.88 was surveyed, to analyse the gene/enzyme network involved in utilization of the plant storage polymer inulin, and of sucrose,...article 2006
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Weenink, X.O. (author), Punt, P.J. (author), van den Hondel, C.A.M.J.J. (author), Ram, A.F.J. (author), TNO Kwaliteit van Leven (author)Although filamentous fungi have a unique property of secreting a large amount of homologous extracellular proteins, the use of filamentous fungi as hosts for the production of heterologous proteins is limited because of the low production levels that are generally reached. Here, we report a general screening method for the isolation of mutants...article 2006
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te Biesebeke, R. (author), Boussier, A. (author), van Biezen, N. (author), van den Hondel, C.A.M.J.J. (author), Punt, P.J. (author), TNO Kwaliteit van Leven (author)Filamentous growth of Aspergillus oryzae on solid cereal substrates involves secretion of substrate converting enzymes and a solid substrate specific polarised hyphal growth phenotype. To identify proteins produced under these specific conditions, the extracts of A. oryzae grown on wheat-based media were analysed using N-terminal sequence...article 2006
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Damveld, R.A. (author), Arentshorst, M. (author), Franken, A. (author), van Kuyk, P.A. (author), Klis, F.M. (author), van den Hondel, C.A.M.J.J. (author), Ram, A.F.J. (author), TNO Kwaliteit van Leven (author)In Aspergillus niger, the genes coding for glutamine:fructose-6-phosphate amidotransferase (gfaA) and α-1,3-glucan synthase (agsA) are induced in response to cell wall stress. In silico analysis of the promoter region of the two genes revealed the presence of putative DNA binding sites for transcription factors involved in stress responses,...article 2005
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te Biesebeke, R. (author), van Biezen, N. (author), de Vos, W.M. (author), van den Hondel, C.A.M.J.J. (author), Punt, P.J. (author), TNO Kwaliteit van Leven (author)Solid-state fermentation (SSF) with Aspergillus oryzae results in high levels of secreted protein. However, control mechanisms of gene expression in SSF have been only poorly studied. In this study we show that both glucoamylase (glaB) and protease (alpA, nptB) genes are highly expressed during surface cultivation on wheat-based solid medium,...article 2005
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te Biesebeke, R. (author), Record, E. (author), van Biezen, N. (author), Heerikhuisen, M. (author), Franken, A. (author), Punt, P.J. (author), van den Hondel, C.A.M.J.J. (author), TNO Kwaliteit van Leven (author)To study the relation between the number of hyphal tips and protein secretion during growth on a solid substrate, we have constructed two mutant strains of Aspergillus oryzae with increased hyphal branching. We have analysed hydrolytic enzyme activities during growth on wheat kernels (WK) of A. oryzae strains carrying the disrupted allele of the...article 2005
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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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te Biesebeke, R. (author), Levin, A. (author), Sagt, C. (author), Bartels, J. (author), Goosen, T. (author), Ram, A. (author), van den Hondel, C. (author), Punt, P. (author), TNO Kwaliteit van Leven (author)Aspergillus oryzae requires polarized growth for colonization of solid substrates, and this growth phenotype differs from that seen in liquid medium. Various experimental approaches were used to identify genes that are differentially expressed when A. oryzae is grown on wheat kernels and in a wheat-based liquid medium. Hybridization of A. oryzae...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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- TNO Kwaliteit van Leven (author), Joosten, V. (author), Gouka, R.J. (author), van den Hondel, C.A.M.J.J. (author), Verrips, C.T. (author), Lokman, B.C. (author) article 2005
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TNO Kwaliteit van Leven (author), Damveld, R.A. (author), Arentshorst, M. (author), van Kuyk, P.A. (author), Klis, F.M. (author), van den Hondel, C.A.M.J.J. (author), Ram, A.F.J. (author)Glycosylphosphatidylinositol (GPI)-anchored proteins in fungi are found at the cell surface, either as plasma membrane proteins (GPI-PMPs) or attached by a remnant of the GPI-anchor to the cell wall (GPI-CWPs). GPI-CWPs can be extracted from the cell wall by treatment with hydrofluoric acid (HF), which cleaves the phosphodiester bond that is...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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Michielse, C.B. (author), Salim, K. (author), Ragas, P. (author), Ram, A.F.J. (author), Kudla, B. (author), Jarry, B. (author), Punt, P.J. (author), van den Hondel, C.A.M.J.J. (author), TNO Voeding (author)Two transformation systems, based on the use of CaCl2/PEG and Agrobacterium tumefaciens, respectively, were developed for the zygomycete Rhizopus oryzae. Irrespective of the selection marker used, a pyr4 marker derived from R. niveus or a dominant amdS+ marker from Aspergillus nidulans, and irrespective of the configuration of the transforming...article 2004