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overproduction of the aspergillus niger feruloyl esterase for pulp bleaching application.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(-1)), improved faea activity 24.5-fold and a yield of 1 g l(-1) of the corresponding protein in the culture medium was achieved. the secreted faea was p ...200312743752
design and production in aspergillus niger of a chimeric protein associating a fungal feruloyl esterase and a clostridial dockerin domain.a chimeric enzyme associating feruloyl esterase a (faea) from aspergillus 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 located upstream of the faea (doc-faea). northern blot analysis showed similar transcript levels for the two constructs, indicating a posttranscriptional bott ...200415574891
cloning and characterization of a tyrosinase gene from the white-rot fungus pycnoporus sanguineus, and overproduction of the recombinant protein in aspergillus niger.a new tyrosinase-encoding gene (2,204 bp) and the corresponding cdna (1,857 nucleotides) from the white-rot fungus pycnoporus sanguineus brfm49 were cloned. this gene consisted of seven exons and six introns and encoded a predicted protein of 68 kda, exceeding the mature tyrosinase by 23 kda. p. sanguineus tyrosinase cdna was over-expressed in aspergillus niger, a particularly suitable fungus for heterologous expression of proteins of biotechnological interest, under the control of the glycerald ...200616151802
construction of engineered bifunctional enzymes and their overproduction in aspergillus niger for improved enzymatic tools to degrade agricultural by-products.two chimeric enzymes, flx and flxlc, were designed and successfully overproduced in aspergillus niger. flx construct is composed of the sequences encoding the feruloyl esterase a (faea) fused to the endoxylanase b (xynb) of a. niger. a c-terminal carbohydrate-binding module (cbm family 1) was grafted to flx, generating the second hybrid enzyme, flxlc. between each partner, a hyperglycosylated linker was included to stabilize the constructs. hybrid proteins were purified to homogeneity, and molec ...200516332795
gene overexpression and biochemical characterization of the biotechnologically relevant chlorogenic acid hydrolase from aspergillus niger.the full-length gene that encodes the chlorogenic acid hydrolase from aspergillus niger cirm brfm 131 was cloned by pcr based on the genome of the strain a. niger cbs 513.88. the complete gene consists of 1,715 bp and codes for a deduced protein of 512 amino acids with a molecular mass of 55,264 da and an acidic pi of 4.6. the gene was successfully cloned and overexpressed in a. niger to yield 1.25 g liter(-1), i.e., 330-fold higher than the production of wild-type strain a. niger cirm brfm131. ...200717630312
gram-scale production of a basidiomycetous laccase in aspergillus niger.we report on the expression in aspergillus niger of a laccase gene we used to produce variants in saccharomyces cerevisiae. grams of recombinant enzyme can be easily obtained. this highlights the potential of combining this generic laccase sequence to the yeast and fungal expression systems for large-scale productions of variants.201423867099
heterologous production of cellobiose dehydrogenases from the basidiomycete coprinopsis cinerea and the ascomycete podospora anserina and their effect on saccharification of wheat straw.cellobiose dehydrogenases (cdhs) are extracellular glycosylated haemoflavoenzymes produced by many different wood-degrading and phytopathogenic fungi. putative cellobiose dehydrogenase genes are recurrently discovered by genome sequencing projects in various phylogenetically distinct fungi. the genomes from the basidiomycete coprinopsis cinerea and the ascomycete podospora anserina were screened for candidate cdh genes, and one and three putative gene models were evidenced, respectively. two put ...201322940800
production of a chimeric enzyme tool associating the trichoderma reesei swollenin with the aspergillus niger feruloyl esterase a for release of ferulic acid.the main goals of this work were to produce the fusion protein of the trichoderma reesei swollenin i (swoi) and aspergillus niger feruloyl esterase a (faea) and to study the effect of the physical association of the fusion partners on the efficiency of the enzyme. the fusion protein was produced up to 25 mg l(-1) in the t. reesei strains rut-c30 and cl847. in parallel, faea alone was produced for use as a control protein in application tests. recombinant faea and swoi-faea were purified to homog ...200616957894
comparison of different fungal enzymes for bleaching high-quality paper pulps.wild and recombinant hydrolases and oxidoreductases with a potential interest for environmentally sound bleaching of high-quality paper pulp (from flax) were incorporated into a totally chlorine free (tcf) sequence that also included a peroxide stage. the ability of feruloyl esterase (from aspergillus niger) and mn2+-oxidizing peroxidases (from phanerochaete chrysosporium and pleurotus eryngii) to decrease the final lignin content of flax pulp was shown. laccase from pycnoporus cinnabarinus (wit ...200415639095
purification and characterization of a chlorogenic acid hydrolase from aspergillus niger catalysing the hydrolysis of chlorogenic acid.among 15 aspergillus strains, aspergillus niger brfm 131 was selected for its high chlorogenic acid hydrolase activity. the enzyme was purified and characterized with respect to its physico-chemical and kinetic properties. four chromatographic steps were necessary to purify the protein to homogeneity with a recovery of 2%. km of the chlorogenic acid hydrolase was estimated to be 10 microm against chlorogenic acid as substrate. under native conditions, the protein presented a molecular mass of 17 ...200515607224
homologous expression of the feruloyl esterase b gene from aspergillus niger and characterization of the recombinant enzyme.the faeb gene encoding the feruloyl esterase b (faeb) was isolated from aspergillus niger brfm131 genomic dna. the faeb gene, with additional sequence coding for a c-terminal histidine tag, was inserted into an expression vector under the control of the gpd promoter and trpc terminator and expressed in a protease deficient a. niger strain. homologous overproduction allows to reach an esterase activity of 18 nkat ml(-1) against mca as substrate. the improvement factor was 16-fold higher as compar ...200415294290
natural and recombinant fungal laccases for paper pulp bleaching.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 oryzae and a. niger hosts using the laci gene from p. cinnabarinus to develop a production process without using the expensive laccase inducers required by t ...200414600793
expression in escherichia coli, refolding and crystallization of aspergillus niger feruloyl esterase a using a serial factorial approach.hydrolysis of plant biomass is achieved by the combined action of enzymes secreted by microorganisms and directed against the backbone and the side chains of plant cell wall polysaccharides. among side chains degrading enzymes, the feruloyl esterase a (faea) specifically removes feruloyl residues. thus, faea has potential applications in a wide range of industrial processes such as paper bleaching or bio-ethanol production. to gain insight into faea hydrolysis activity, we solved its crystal str ...200717533138
production of toxic metabolites in aspergillus niger, aspergillus oryzae, and trichoderma reesei: justification of mycotoxin testing in food grade enzyme preparations derived from the three fungi.aspergillus niger, aspergillus oryzae, and trichoderma reesei are three important production organisms used in industrial fermentations. several of the fungal secondary metabolites produced by selected strains of these three fungi are capable of eliciting toxicity in animals. among those toxic substances are the well-known mycotoxins 3-nitropropionic acid and ochratoxin a. however, many others, such as kojic acid, may not be true mycotoxins. the production, extraction, chemical structure, and th ...200415041150
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