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Conesa, A. (author), van den Hondel, C.A.M.J.J. (author), Punt, P.J. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)To get insight into the limiting factors existing for the efficient production of fungal peroxidase in filamentous fungi, the expression of the Phanerochaete chrysosporium lignin peroxidase H8 (lipA) and manganese peroxidase (MnP) H4 (mnp1) genes in Aspergillus niger has been studied. For this purpose, a protease-deficient A. niger strain and...article 2000
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Krasevec, N. (author), van de Hondel, C.A.M.J.J. (author), Komel, R. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)A gene-fusion expression strategy was applied for heterologous expression of human lymphotoxin alpha (LTα) in the Aspergillus niger AB1.13 protease-deficient strain. The LTα gene was fused with the A. niger glucoamylase GII-form as a carrier-gene, behind its transcription control and secretion signals. Special attention was paid to the influence...article 2000
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van den Brink, J.M. (author), Punt, P.J. (author), van Gorcom, R.F.M. (author), van den Hondel, C.A.M.J.J. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)Cytochrome P450 enzyme systems are found throughout nature and are involved in many different, often complex, bioconversions. In the endoplasmic reticulum of the filamentous fungus Aspergillus niger a cytochrome P450 enzyme system is present that is capable of the para-hydroxylation of benzoate. The expression of the two genes encoding the...article 2000
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Krasevec, N. (author), van den Hondel, C.A.M.J.J. (author), Komel, R. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)A gene-fusion expression strategy was applied for the heterologous expression of hTNF-α in A. niger AB1.13. The TNF-α gene was fused with the A. niger glucoamylase GII form as a carrier-gene, behind its transcription control and secretion signal. The protein was expressed in the cells in the form of a glucoamylase-fusion protein, but was not...article 2000
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TNO Voeding (author), Gordon, C.L. (author), Khalaj, V. (author), Ram, A.F.J. (author), Archer, D.B. (author), Brookman, J.L. (author), Trinci, A.P.J. (author), Jeenes, D.J. (author), Doonan, J.H. (author), Wells, B. (author), Punt, P.J. (author), van den Hondel, C.A.M.J.J. (author), Robson, G.D. (author)A glucoamylase::green fluorescent protein fusion (GLA::sGFP) was constructed which allows the green fluorescent protein to be used as an in vivo reporter of protein secretion in Aspergillus niger. Two secretory fusions were designed for secretion of GLA::sGFP which employed slightly different lengths of the glucoamylase protein (GLA499 and...article 2000
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Gordon, C.L. (author), Khalaj, V. (author), Ram, A.F.J. (author), Archer, D.B. (author), Brookman, J.L. (author), Trinci, A.P.J. (author), Jeenes, D.J. (author), Doonan, J.H. (author), Wells, B. (author), Punt, P.J. (author), van den Hondel, C.A.M.J.J. (author), Robson, G.D. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)A glucoamylase: :green fluorescent protein fusion (GLA: :sGFP) was constructed which allows the green fluorescent protein to be used as an in vivo reporter of protein secretion in Aspergillus niger. Two secretory fusions were designed for secretion of GLA: :sGFP which employed slightly different lengths of the glucoamylase protein (GLA499 and...article 2000
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Centraal Instituut voor Voedingsonderzoek TNO TNO Voeding (author), Ngiam, C. (author), Jeenes, D.J. (author), Punt, P.J. (author), van den Hondel, C.A.M.J.J. (author), Archer, D.B. (author)Protein disulfide isomerase (PDI) is important in assisting the folding and maturation of secretory proteins in eukaryotes. A gene, pdiA, encoding PDIA was previously isolated from Aspergillus niger, and we report its functional characterization here. Functional analysis of PDIA showed that it catalyzes the refolding of denatured and reduced...article 2000
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Santerre Henriksen, A.L. (author), Even, S. (author), Müller, C. (author), Punt, P.J. (author), van den Hondel, C.A.M.J.J. (author), Nielsen, J. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)An Aspergillus niger strain expressing a red-shifted green fluorescent protein (GFP) in the cytoplasm under the control of the glucoamylase promoter (PglaA) was characterized with respect to its physiology and morphology. Although xylose acted as a repressor carbon source during batch cultivations, PglaA-driven GFP expression by the glucoamylase...article 1999
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Zhu, Y. (author), Knol, W. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)To explore the possibilities of using lupine as a soybean replacement in fermented foods, fermentation profiles of lupine and soybean by Aspergillus oryzae and A. sojae, respectively, in a solid-state culture were compared. Biomass, spore concentration, oxygen consumption rate, carbon dioxide production rate, water activity, and production of...article 1998
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Randerath, K. (author), Sriram, P. (author), Moorthy, B. (author), Aston, J.P. (author), Baan, R.A. (author), van den Berg, P.T.M. (author), Booth, E.D. (author), Watson, W.P. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)32P-postlabelling analysis for detecting DNA adducts formed by polycyclic aromatic compounds is one of the most widely used techniques for assessing genotoxicity associated with these compounds. In cases where the formation of adducts is extremely low, a crucial step in the analysis is an enrichment procedure for adducts prior to the...article 1998
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van Gemeren, I.A. (author), Beijersbergen, A. (author), van den Hondel, C.A.M.J.J. (author), Verrips, C.T. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)Several cutinase variants derived by molecular modelling and site- directed mutagenesis of a cutinase gene from Fusarium solani pisi are poorly secreted by Saccharomyces cerevisiae. The majority of these variants are successfully produced by the filamentous fungus Aspergillus awamori. However, the L51S and T179Y mutations caused reductions in...article 1998
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van Zeijl, C.M.J. (author), van de Kamp, E.H.M. (author), Punt, P.J. (author), Selten, G.C.M. (author), Hauer, B. (author), van Gorcom, R.F.M. (author), van den Hondel, C.A.M.J.J. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)A method was developed to perform PCR directly on mycelial pellets or colonies treated with NOVOzym 234. The method allows rapid screening of large numbers of transformants of both sporulating and non-sporulating fungi for the presence of (co)transforming plasmid copies or for specific genetic modifications such as gene disruption and site...article 1998
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- Frenken, L.G.J. (author), Hessing, J.G.M. (author), van den Hondel, C.A.M.J.J. (author), Verrips, C.T. (author), Centraal Instituut voor Voedingsonderzoek TNO (author) article 1998
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Withers, J.M. (author), Swift, R.J. (author), Wiebe, M.G. (author), Robson, G.D. (author), Punt, P.J. (author), van den Hondel, C.A.M.J.J. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)When grown on a medium containing 5 g maltodextrin L-1, Aspergillus niger transformant N402[pAB6-10]B1, which has an additional 20 copies of the glucoamylase (glaA) gene, produced 320 ± 8 mg (mean ± S.E.) glucoamylase (GAM) L-1 in batch culture and 373 ± 9 mg GAM L-1 in maltodextrin- limited chemostat culture at a dilution rate of 0.13 h-1....article 1998
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Punt, P.J. (author), van Gemeren, I.A. (author), Drint-Kuijvenhoven, J. (author), Hessing, J.G.M. (author), Muijlwijk van - Harteveld, G.M. (author), Beijersbergen, A. (author), Verrips, C.T. (author), van den Hondel, C.A.M.J.J. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)The function of the endoplasmic-reticulum-localized chaperone binding protein (BiP) in relation to protein secretion in filamentous fungi was studied. It was shown that the overproduction of several homologous and heterologous recombinant proteins by Aspergillus strains induces the expression of bipA, the BiP-encoding gene from Aspergillus niger...article 1998
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Gouka, R.J. (author), Punt, P.J. (author), van den Hondel, C.A.M.J.J. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)In this study we have analyzed the effects of a glucoamylase gene fusion on the mRNA levels and protein levels for the human interleukin-6 gene (hil6) and the guar α-galactosidase gene (aglA). Previously it was shown that production of nonfused α-galactosidase and hIL-6 in Aspergillus awamori was limited at transcriptional and (post...article 1997
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Gouka, R.J. (author), Stam, H. (author), Fellinger, A.J. (author), Muijsenberg, R.J.G.T. (author), van de Wijngaard, A.J. (author), Punt, P.J. (author), Musters, W. (author), van den Hondel, C.A.M.J.J. (author), TNO Voeding (author)In this study, induction and repression kinetics of the expression of the Aspergillus awamori 1,4-β-endoxylanase A (exlA) gene under defined physiological conditions was analyzed at the mRNA and the protein levels. Induction was analyzed by pulsing D-xylose to a sucrose-limited continuous culture of an A. awamori 1,4-β-endoxylanase A (EXLA)...article 1996
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van den Brink, H.J.M. (author), van Nistelrooy, H.J.G.M. (author), de Waard, M.A. (author), van den Hondel, C.A.M.J.J. (author), van Gorcom, R.F.M. (author), Prins Maurits Laboratorium TNO (author)In this paper we describe the effects of over-expression of the Penicillium italicum gene encoding eburicol 14α-demethylase (cyp51), in Aspergillus niger strains with one or multiple copies of the gene encoding cytochrome P450 reductase (cpr A), on the eburicol 14α-demethylase activity. Eburicol 14α-demethylase activity was determined by...article 1996
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Gouka, R.J. (author), Punt, P.J. (author), Hessing, J.G.M. (author), van den Hondel, C.A.M.J.J. (author), TNO Voeding (author)A study was carried out to obtain more insight into the parameters that determine the secretion of heterologous proteins from filamentous fungi. A strategy was chosen in which the mRNA levels and protein levels of a number of heterologous genes of different origins were compared. All genes were under control of the Aspergillus awamori 1,4-β...article 1996
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TNO Nutrition and Food Research (author), van Gemeren, I.A. (author), Beijersbergen, A. (author), Musters, W. (author), Gouka, R.J. (author), van den Hondel, C.A.M.J.J. (author), Verrips, C.T. (author)A synthetic derivative of the cutinase cDNA of Fusarium solani pisi was expressed in Aspergillus awamori using the A. awamori endoxylanase II (exlA) promoter and terminator. The influence of the origin of the pre-sequence and the presence of a pro-sequence on the efficiency of extracellular cutinase production was analyzed in single-copy...article 1996