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Aspergillus niger genome-wide analysis reveals a large number of novel alpha-glucan acting enzymes with unexpected expression profiles

Yuan, X.-L. (author), van der Kaaij, R.M. (author), van den Hondel, C.A.M.J.J. (author), Punt, P.J. (author), van der Maarel, M.J.E.C. (author), Dijkhuizen, L. (author), Ram, A.F.J. (author), TNO Kwaliteit van Leven (author)

The filamentous ascomycete Aspergillus niger is well known for its ability to produce a large variety of enzymes for the degradation of plant polysaccharide material. A major carbon and energy source for this soil fungus is starch, which can be degraded by the concerted action of α-amylase, glucoamylase and α-glucosidase enzymes, members of the...

article 2008

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A novel screening method for cell wall mutants in Aspergillus niger identifies UDP-galactopyranose mutase as an important protein in fungal cell wall biosynthesis

Damveld, R.A. (author), Franken, A. (author), Arentshorst, M. (author), Punt, P.J. (author), Klis, F.M. (author), van den Hondel, C.A.M.J.J. (author), Ram, A.F.J. (author), TNO Kwaliteit van Leven (author)

To identify cell wall biosynthetic genes in filamentous fungi and thus potential targets for the discovery of new antifungals, we developed a novel screening method for cell wall mutants. It is based on our earlier observation that the Aspergillus niger agsA gene, which encodes a putative a-glucan synthase, is strongly induced in response to...

article 2008

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Characterization of the Aspergillus niger prtT, a unique regulator of extracellular protease encoding genes

Punt, P.J. (author), Schuren, F.H.J. (author), Lehmbeck, J. (author), Christensen, T. (author), Hjort, C. (author), van den Hondel, C.A.M.J.J. (author), TNO Kwaliteit van Leven (author)

Expression of several Aspergillus niger genes encoding major secreted, but not vacuolar, protease genes including the major acid protease gene pepA, was shown to be affected in the previously isolated A. niger protease mutant, AB1.13 [Mattern, I.E., van Noort, J.M., van den Berg, P., Archer, D.A., Roberts, I.N., Hondel, C.A.M.J.J., 1992....

article 2008

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Two novel, putatively cell wall-associated and glycosylphosphatidylinositol-anchored α-glucanotransferase enzymes of Aspergillus niger

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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Phylogenetic and biochemical characterization of a novel cluster of intracellular fungal α-amylase enzymes

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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Identification of secreted proteins of Aspergillus oryzae associated with growth on solid cereal substrates

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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Branching mutants of Aspergillus oryzae with improved amylase and protease production on solid substrates

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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Production of bifunctional proteins by Aspergillus awamori: Llama variable heavy chain antibody fragment (VHH) R9 coupled to Arthromyces ramosus peroxidase (ARP)

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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Food allergy: What do we learn from animal models?

Knippels, L.M.J. (author), van Wijk, F. (author), Penninks, A.H. (author), TNO Voeding (author)

Purpose of review This review summarizes selected articles on animal models of food allergy published in 2003. The research areas that are covered include mechanistic studies, the search for new therapies, as well as screening models for hazard identification of potential allergens. Recent findings Novel treatment options of both prevention and...

article 2004

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Endoplasmic reticulum stress leads to the selective transcriptional downregulation of the glucoamylase gene in Aspergillus niger

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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Design and production in Aspergillus niger of a chimeric protein associating a fungal feruloyl esterase and a clostridial dockerin domain

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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The role of the Aspergillus niger furin-type protease gene in processing of fungal proproteins and fusion proteins: Evidence for alternative processing of recombinant (fusion-) proteins

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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Is Candida albicans a trigger in the onset of coeliac disease?

Nieuwenhuizen, W.F. (author), Pieters, R.H.H. (author), Knippels, L.M.J. (author), Jansen, M.C.J.F. (author), Koppelman, S.J. (author)

Coeliac disease is a T-cell-mediated autoimmune disease of the small intestine that is induced by ingestion of gluten proteins from wheat, barley, or rye. We postulate that Candida albicans is a trigger in the onset of coeliac disease. The virulence factor of C albicans - hyphal wall protein 1 (HWP1) - contains aminoacid sequences that are...

article 2003

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Calnexin overexpression increases manganese peroxidase production in Aspergillus niger

TNO Voeding Centraal Instituut voor Voedingsonderzoek TNO (author), Conesa, A. (author), Jeenes, D. (author), Archer, D.B. (author), van den Hondel, C.A.M.J.J. (author), Punt, P.J. (author)

Heme-containing peroxidases from white rot basidiomycetes, in contrast to most proteins of fungal origin, are poorly produced in industrial filamentous fungal strains. Factors limiting peroxidase production are believed to operate at the posttranslational level. In particular, insufficient availability of the prosthetic group which is required...

article 2002

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The secretion pathway in filamentous fungi: A biotechnological view

Conesa, A. (author), Punt, P.J. (author), van Luijk, N. (author), van den Hondel, C.A.M.J.J. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)

The high capacity of the secretion machinery of filamentous fungi has been widely exploited for the production of homologous and heterologous proteins; however, our knowledge of the fungal secretion pathway is still at an early stage. Most of the knowledge comes from models developed in yeast and higher eukaryotes, which have served as reference...

article 2001

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Identification and characterization of a family of secretion-related small GTPase-encoding genes from the filamentous fungus Aspergillus niger: A putative SEC4 homologue is not essential for growth

Punt, P.J. (author), Seiboth, B. (author), Weenink, X.O. (author), van Zeijl, C. (author), Lenders, M. (author), Konetschny, C. (author), Ram, A.F.J. (author), Montijn, R. (author), Kubicek, C.P. (author), van den Hondel, C.A.M.J.J. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)

DNA fragments containing genetic information for five secretion-related small GTPases of Aspergillus niger (srgA-E) were isolated and identified as members of different Rab/Ypt subfamilies. This isolation and the search for similar sequences in fungal genomic and EST databases showed that, in contrast to Saccharomyces cerevisiae, filamentous...

article 2001

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Expression of the Caldariomyces fumago chloroperoxidase in Aspergillus niger and characterization of the recombinant enzyme

Centraal Instituut voor Voedingsonderzoek TNO TNO Voeding (author), Conesa, A. (author), van de Velde, F. (author), van Rantwijk, F. (author), Sheldon, R.A. (author), van den Hondel, C.A.M.J.J. (author), Punt, P.J. (author)

The Caldariomyces fumago chloroperoxidase was successfully expressed in Aspergillus niger. The recombinant enzyme was produced in the culture medium as an active protein and could be purified by a three-step purification procedure. The catalytic behavior of recombinant chloroperoxidase (rCPO) was studied and compared with that of native CPO. The...

article 2001

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Characterization of a foldase, protein disulfide isomerase A, in the protein secretory pathway of Aspergillus niger

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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Localization of synthesis of β1,6-glucan in Saccharomyces cerevisiae

Montijn, R.C. (author), Vink, E. (author), Müller, W.H. (author), Verkleij, A.J. (author), van den Ende, H. (author), Henrissat, B. (author), Klis, F.M. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)

β1,6-Glucan is a key component of the yeast cell wall, interconnecting cell wall proteins, β1,3-glucan, and chitin. It has been postulated that the synthesis of β1,6-glucan begins in the endoplasmic reticulum with the formation of protein-bound primer structures and that these primer structures are extended in the Golgi complex by two putative...

article 1999

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Filamentous fungi as production organisms for glycoproteins of bio-medical interest

Maras, M. (author), van Die, I. (author), Contreras, R. (author), van den Hondel, C.A.M.J.J. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)

Filamentous fungi are commonly used in the fermentation industry for large scale production of glycoproteins. Several of these proteins can be produced in concentrations up to 20-40 g per litre. The production of heterologous glycoproteins is at least one or two orders of magnitude lower but research is in progress to increase the production...

article 1999