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cloning and heterologous expression of gene encoding a polygalacturonase from aspergillus awamori.a polygalacturonase gene of aspergillus awamori ifo 4033 was cloned by genomic southern hybridization with a probe of a dna fragment synthesized by pcr. this was done using primers constructed based on the n-terminal amino acid sequence of a polygalacturonase, protopectinase-as, produced by the strain and the consensus internal amino acid sequence of fungal polygalacturonases. the cloned polygalacturonase gene, containing an orf, encodes 362 amino acids, including a 52-bp intron. it contains the ...200010993142
[characterization and transformation of anti-foa strain of aspergillus awamori].a suspension of freshly harvested spores of aspergillus awamori sg1 strain was exposed to uv light. the irradiated spores with survival rate of 10%-15% were plated directly on minimal medium (mm) agar plates containing 10 mmol/l uridine and 5-fluoro-orotic acid (foa 1 mg/ml). five stable anti-foa colonies with the reversed rate below 10(-9) were obtained. all anti-foa strains, can grow on uridine or uricil-containing minimal medium agar plate but not on orotic acid-containing mm plate, indicatin ...200012548939
xylanase production by aspergillus awamori in solid-state fermentation and influence of different nitrogen sources.the use of purified xylan as a substrate for bioconversion into xylanases increases the cost of enzyme production. consequently, there have been attempts to develop a bioprocess to produce such enzymes using different lignocellulosic residues. filamentous fungi have been widely used to produce hydrolytic enzymes for industrial applications, including xylanases, whose levels in fungi are generally much higher than those in yeast and bacteria. considering the industrial importance of xylanases, th ...200111963896
production of beta-mannanase and beta-mannosidase from aspergillus awamori k4 and their properties.beta-mannanase and beta-mannosidase from aspergillus awamori k4 was produced by solid culture with coffee waste and wheat bran. the optimum composition for enzyme production was 40% coffee waste-60% wheat bran. two enzymes were partially purified. optimum ph was about 5 for both enzymes, and optimum temperature was around 80 degrees c for beta-mannanase and 60-70 degrees c for beta-mannosidase. these enzymes produced some oligosaccharides from glucomannan and galactomannan by their hydrolyzing a ...200111381326
identification, classification and phylogeny of the aspergillus section nigri inferred from mitochondrial cytochrome b gene.the partial mitochondrial cytochrome b gene from 32 strains of 12 species belonging to aspergillus section nigri was amplified by the polymerase chain reaction and sequenced directly. using 402 nucleotide characters, nucleotide-based and amino acid-based phylogenetic trees were inferred and the genetic divergence among the species was evaluated. based on analyses of the 402-bp nucleotide and 133-amino acid sequences, strains were divided into 11 dna types and five amino acid types. aspergillus n ...200111425482
factors affecting the production of a single-chain antibody fragment by aspergillus awamori in a stirred tank reactor.a recombinant strain of aspergillus awamori expressing anti-lysozyme single chain antibody fragments (scfv), under the control of a xylanase promoter, was studied in order to investigate the impact of medium, induction regime and protease production on the expression of the product. experiments with the time of induction showed that the optimum results are achieved when induction is started in the late exponential phase (21 h after inoculation) improving the titer of the product from 14.5 mg l(- ...200111485420
fungal flora of the digestive tract of 5 species of triatomines vectors of trypanosoma cruzi, chagas 1909.a study of the mycobiota in the digestive tract of 5 important species of triatomines, triatoma brasiliensis, t infestans, t. sordida, t. pseudomaculata and t. vitticeps, was made. the digestive tracts of 164 adults and 535 nymphs of those triatomines were studied and 393 fungal strains were isolated. the genera with the greatest number of species were penicillium (19 species), aspergillus (17 species) and acremonium (5 species) and the most frequent species, in decreasing order, were penicilliu ...200111502063
crystal structure of maltose phosphorylase from lactobacillus brevis: unexpected evolutionary relationship with glucoamylases.maltose phosphorylase (mp) is a dimeric enzyme that catalyzes the conversion of maltose and inorganic phosphate into beta-d-glucose-1-phosphate and glucose without requiring any cofactors, such as pyridoxal phosphate. the enzyme is part of operons that are involved in maltose/malto-oligosaccharide metabolism. maltose phosphorylases have been classified in family 65 of the glycoside hydrolases. no structure is available for any member of this family.200111587643
ethanol production from starch by immobilized aspergillus awamori and saccharomyces pastorianus using cellulose carriers.a simultaneous saccharification and fermentation (ssf) process was investigated to produce ethanol using two kinds of cellulose carriers that were respectively suitable for immobilization of aspergillus awamori and saccharomyces pastorianus. the maximum ethanol concentration attained by the batch operation was 25.5 g l(-1). under suitable conditions, both cellulose carriers with immobilized cells could be reused efficiently for three cycles. the total amount of ethanol production was 66.0 g (per ...200111598811
a defined level of protein disulfide isomerase expression is required for optimal secretion of thaumatin by aspegillus awamori.thaumatin, a 22-kda protein containing eight disulfide bonds, is secreted by the filamentous fungus aspergillus awamori at levels which are dependent upon the extent of overexpression of protein disulfide isomerase (pdia). additional copies of the pdia-encoding gene pdia were introduced into a strain of a. awamori that expresses a cassette encoding thaumatin. transformants with different levels of pdia mrna and measured pdia levels were chosen for examination of the impact that pdia levels had o ...200111683266
[immobilization of glucoamylase from aspergillus awamori 466, and properties of the preparation].glucoamylase from aspergillus awamori 466 was immobilized on various supports. the enzyme sorption depends on its amount, the type of support, and immobilization conditions. the kinetics of acidic inactivation of the native and immobilized enzyme was studied. the immobilized enzyme was more resistant to temperature and ph. the mechanism of the enzyme binding to the support was investigated by ir spectroscopy.200111357426
cloning of a phenol oxidase gene from acremonium murorum and its expression in aspergillus awamori.fungal multicopper oxidases have many potential industrial applications, since they perform reactions under mild conditions. we isolated a phenol oxidase from the fungus acremonium murorum var. murorum that was capable of decolorizing plant chromophores (such as anthocyanins). this enzyme is of interest in laundry-cleaning products because of its broad specificity for chromophores. we expressed an a. murorum cdna library in saccharomyces cerevisiae and subsequently identified enzyme-producing ye ...200111375170
effect of lactoferrin on helicobacter felis induced gastritis.lactoferrin possesses antibiotic, antiinflammatory, and immune-modulating properties that may be active against the gastritis-, ulcer- and cancer-inducing bacterium helicobacter pylori. in vitro testing of bovine and human lactoferrin by several laboratories has shown significant bacteriostatic and bactericidal activity. subsequent in vivo testing of bovine lactoferrin in animal models of h. pylori infection has shown beneficial effects of this agent. our laboratory has utilized a mouse model th ...200211908634
aminoadipate reductase gene: a new fungal-specific gene for comparative evolutionary analyses.in fungi, aminoadipate reductase converts 2-aminoadipate to 2-aminoadipate 6-semialdehyde. however, other organisms have no homologue to the aminoadipate reductase gene and this pathway appears to be restricted to fungi. in this study, we designed degenerate primers for polymerase chain reaction (pcr) amplification of a large fragment of the aminoadipate reductase gene for divergent fungi.200211931673
constitutive expression of the trichoderma reesei beta-1,4-xylanase gene (xyn2) and the beta-1,4-endoglucanase gene (egl) in aspergillus niger in molasses and defined glucose media.the xylanase ii (xyn2)- and endoglucanase i (egl)-encoding regions of trichoderma reesei qm6a were successfully expressed in aspergillus niger d15 under the transcriptional control of the glyceraldehyde-6-phosphate dehydrogenase (gpd) promoter from a. niger and the glaa terminator of aspergillus awamori. a stable xyn2 transformant produced beta-xylanase activity of 8,000 nkat/ml and 5,000 nkat/ml in shake-flask cultures containing defined or 20% (v/v) molasses medium, respectively. the recombina ...200211954792
polygalacturonase production by aspergillus awamori on wheat in solid-state fermentation.the production of exo-polygalacturonase (exo-pg) and endo-pg by aspergillus awamori grown on wheat in solid-state fermentation was studied. endo- and exo-pg activities were detected after 24 h of inoculation. glucose released from starch hydrolysis acted as a catabolite repressor for the exo-pg enzyme. in contrast, endo-pg production was not affected by glucose repression. when milled grains were used, the particle-size distribution and the chemical composition of the medium influenced the rate ...200211876407
purification, characterization, gene cloning and preliminary x-ray data of the exo-inulinase from aspergillus awamori.extracellular exo-inulinase has been isolated from a solid-phase culture of the filamentous fungus aspergillus awamori var. 2250. the apparent molecular mass of the monomer enzyme was 69 +/- kda, with a pi of 4.4 and a ph optimum of 4.5. the enzyme hydrolysed the beta-(2-->1)-fructan (inulin) and beta-(2-->6)-fructan (levan) via exo-cleavage, releasing fructose. the values for the michaelis constants k(m) and v(max) in the hydrolysis of inulin were 0.003 +/- 0.0001 mm and 175 +/- 5 micromol.min( ...200211829749
expression of an alpha-galactosidase from saccharomyces cerevisiae in aspergillus awamori and aspergillus oryzae.a gene encoding alpha-galactosidase activity was isolated by polymerase chain reaction (pcr) from saccharomyces cerevisiae ncyc686 and separately placed under the control of transcriptional elements regulating alpha-amylase expression in aspergillus oryzae and glucoamylase expression in a. awamori. following transformation of both a. oryzae and a. awamori with their respective expression vectors, induction of heterologous alpha-galactosidase from positively selected clones was effected through t ...200212074058
silencing of the aspergillopepsin b (pepb) gene of aspergillus awamori by antisense rna expression or protease removal by gene disruption results in a large increase in thaumatin production.aspergillopepsin b was identified in culture broths of aspergillus awamori by in situ detection of its proteolytic activity and by immunodetection with anti-aspergillopepsin b antibodies. severe thaumatin degradation was observed after in vitro treatment of thaumatin with purified aspergillopepsin b. the pepb gene encoding aspergillopepsin b of a. awamori was cloned and characterized. it is located in chromosome iv of a. awamori, as shown by pulsed-field gel electrophoresis, and encodes a protei ...200212089041
alcohol production from starch by mixed cultures of aspergillus awamori and immobilized saccharomyces cerevisiae at different agitation speeds.the influence of different agitation speeds on alcoholic fermentation by free and immobilized cells of saccharomyces cerevisiae in a co-culture with free cells of aspergillus awamori was investigated. starch hydrolysis and glucose, glucoamylase and alpha-amylase accumulation in the fermentation medium was affected by agitation speed. maximum amounts of the enzymes were obtained at 150-200 rpm after 72 h where, maximum growth and glucose accumulations were noticed at 200-300 rpm. alcohol producti ...200212111743
ultra scaledown to predict filtering centrifugation of secreted antibody fragments from fungal broth.extracellularly expressed anti-hen egg lysozyme single-chain antibody fragments (scfv) produced by aspergillus awamori were recovered using filtering centrifugation. two filtering centrifuges with 0.5- and 30-l capacities were used to represent laboratory- and pilot-scale equipment, respectively. critical regime analysis using the computational fluid dynamics (cfd) technique provided information about the local energy dissipation rates in both units. experimental data indicated loss of scfv acti ...200212115401
swollenin, a trichoderma reesei protein with sequence similarity to the plant expansins, exhibits disruption activity on cellulosic materials.plant cell wall proteins called expansins are thought to disrupt hydrogen bonding between cell wall polysaccharides without hydrolyzing them. we describe here a novel gene with sequence similarity to plant expansins, isolated from the cellulolytic fungus trichoderma reesei. the protein named swollenin has an n-terminal fungal type cellulose binding domain connected by a linker region to the expansin-like domain. the protein also contains regions similar to mammalian fibronectin type iii repeats, ...200212199698
evaluation of filamentous fungi and inducers for the production of endo-polygalacturonase by solid state fermentation.aspergillus oryzae cct 3940, aspergillus awamori nrrl 3112 and a trichoderma sp.) were compared for their capacity to produce endo-polygalacturonase (endo-pg) in solid state fermentation. maximum pectinolytic activity was reached in 72 h of growth, the best two fungal strains being a. niger t0005007-2 and a. oryzae cct 3940. three types of commercial purified pectin and four of unprocessed pectin (tangerine, orange, tahiti lime and sweet lime rind) were used to assess the effect of pectin on the ...200212240994
aspergillin pz, a novel isoindole-alkaloid from aspergillus awamori.aspergillin pz was obtained from the fermentation of aspergillus awamori (nakazawa) by activity-guided fractionation and purification. its structure was elucidated on the basis of spectral data, especially by 2d nmr, and finally confirmed by an x-ray analysis. it could induce conidia of p. oryzae to deform moderately.200212374381
engineering of polyploid saccharomyces cerevisiae for secretion of large amounts of fungal glucoamylase.we engineered saccharomyces cerevisiae cells that produce large amounts of fungal glucoamylase (gai) from aspergillus awamori var. kawachi. to do this, we used the delta-sequence-mediated integration vector system and the heat-induced endomitotic diploidization method. delta-sequence-mediated integration is known to occur mainly in a particular chromosome, and the copy number of the integration is variable. in order to construct transformants carrying the gai gene on several chromosomes, haploid ...200212406766
effects of root-dip treatment with certain phosphate solubilizing microorganisms on the fusarial wilt of tomato.root-dip application of bacillus subtilis, pseudomonas fluorescens, aspergillus awamori, aspergillus niger and penicillium digitatum resulted in significant decline in the rhizosphere population of fusarium oxysporum f. sp. lycopersici. a significant decrease in the severity of wilt occurred with a. awamori (37.1%) and p. digitatum (21.3%) compared to the control. root-dip treatment with the phosphate solubilizing microorganisms tested resulted in significant increase in the yield of tomato, bei ...200212227549
molecular dynamics simulations of the unfolding mechanism of the catalytic domain from aspergillus awamori var. x100 glucoamylase.in this study, 200 ps molecular dynamics simulations were conducted to investigate the unfolding mechanism of the catalytic domain of glucoamylase from aspergillus awamori var. x100. the unfolding of this domain was suggested to follow a putative hierarchical manner, in which the heavily o-glycosylated belt region from residues t440 to a471 acted as the initiation site, followed by the alpha-helix secondary structure destruction, and then the collapse of the catalytic center pocket. the o-glycos ...200312529155
cloning and expression of fungal phytases in genetically modified strains of aspergillus awamori.in an effort to produce phytases cost-effectively, and to determine the efficiency of a novel expression system, the genes for aspergillus awamori ( phya) phytase and aspergillus fumigatus ( phya) phytase (a putative thermostable enzyme) were cloned and overexpressed in a. awamori. regulation of phytase expression was achieved by separately placing the genes under the transcriptional control of the glucoamylase a ( glaa) promoter of a. awamori. a gene fusion strategy was employed that involved t ...200314513384
[effect of various sources of nitrogen and carbon on the biosynthesis of proteolytic enzymes in a culture of aspergillus awamori 21/96].effects of various conditions of nitrogen and carbon nutrition on the biosynthesis of proteolytic enzymes in a selected culture of aspergillus awamori 21/96 were studied. this strain was shown to produce proteolytic enzymes constitutively. in the presence of mineral sources of nitrogen, the synthesis of the enzymes under study was not induced by proteinaceous substrates. optimum conditions of the enzyme biosynthesis were achieved with casein as a source of nitrogen and starch or dulcitol as a so ...200312722657
influence of carbon, nitrogen and phosphorous sources on glucoamylase production by aspergillus awamori in solid state fermentation.it was the objective of the present study to increase the production of glucoamylase by aspergillus awamori through solid state fermentation, using wheat bran as the main carbon source and (nh4)2so4, urea, kh2po4, glucose, maltose and starch as additional nitrogen, phosphorus, and carbon sources. the production of glucoamylase is strongly influenced by n and c sources. a 100% increase was observed when the (nh4)2so4 was replaced by urea, with c/n = 4.8, using maltose as the additional carbon sou ...200314577636
predicted unfolding order of the 13 alpha-helices in the catalytic domain of glucoamylase from aspergillus awamori var. x100 by molecular dynamics simulations.the unfolding mechanism of the 13 alpha-helices in the catalytic domain of aspergillus awamori var. x100 glucoamylase was investigated by 200 ps molecular dynamics simulations in explicit water with temperature jump technique. rather than a simultaneous event, the unfolding of these 13 alpha-helices followed a random ordered mechanism as alpha8-->alpha1-->alpha11-->alpha7-->alpha10-->alpha3-->alpha12-->alpha13-->alpha4-->alpha5-->alpha9-->alpha6-->alpha2. no significant relationships were found ...200314524723
growth and pectinase production by aspergillus mexican strain protoplast regenerated under acidic stress.protoplasts from aspergillus sp. fp-180 and aspergillus awamori nrrl- 3112 were released and regenerated at extreme acidic conditions. the best conditions for protoplast release were 0.8 m kci, ph 5.8, and 3 h of digestion using mycelia from 12- to 16-h cultures from either aspergillus sp. fp-180 or a. awamori nrrl-3112. the addition of fresh mycelia to an ongoing digestion after 1 h increased protoplast 4.5-5 times. a regeneration efficiency of 90% was attained at ph 6.0, and it was possible to ...200314566066
genetically modified soybeans: false-positive detection in fermented natural soybean (tempe).tempe was prepared using mixtures of natural soybean and genetically modified roundup ready (rur) soybean fermented with natural rhizopus sp. the amount of rur soybean was quantified using an elisa plate test. the rur signal decreased during fermentation. in the control experiments on fermentation of non-rur soybean, the tempe gave a false-positive rur signal. the cross-reacting substance was generated only in non-rur soybean during fermentation by rhizopus sp., rhizopus oligosporus, r. oryzae, ...200314571970
protein engineering to improve the thermostability of glucoamylase from aspergillus awamori based on molecular dynamics simulations.twelve mutations were constructed to improve the thermostability of glucoamylase from aspergillus awamori based on the results of molecular dynamics simulations. the thermal unfolding of the catalytic domain followed a putative hierarchical behavior. in addition, the unfolding of the 13 alpha-helices obeyed the random ordered mechanism, in which the alpha-helices 8, 1 and 11 unfolded more rapidly than the others. the catalytic center was well protected by the (alpha/alpha)(6)-barrel at simulatio ...200312646689
a novel circulating loop bioreactor with cells immobilized in loofa ( luffa cylindrica) sponge for the bioconversion of raw cassava starch to ethanol.a circulating loop bioreactor (clb) with cells immobilized in loofa sponge was constructed for simultaneous aerobic and anaerobic processes. the clb consists of an aerated riser and a non-aerated downcomer column connected at the top and bottom by cylindrical pipes. ethanol production from raw cassava starch was investigated in the clb. aspergillus awamori iam 2389 and saccharomyces cerevisiae ir2 immobilized on loofa sponge were placed, respectively, in the aerated riser column and non-aerated ...200312664145
l-lactic acid production from raw cassava starch in a circulating loop bioreactor with cells immobilized in loofa (luffa cylindrica).l-lactic acid was produced from raw cassava starch, by simultaneous enzyme production, starch saccharification and fermentation in a circulating loop bioreactor with aspergillus awamori and lactococcus lactis spp. lactis immobilized in loofa sponge. a. awamori was immobilized directly in cylindrical loofa sponge while the l. lactis was immobilized in a loofa sponge alginate gel cube. in the loofa sponge alginate gel cube, the sponge serves as skeletal support for the gel with the cells. the algi ...200312889820
evaluation of alternative reporter genes for the yeast two-hybrid system.the yeast two-hybrid system is a powerful tool for screening protein-protein interactions and has also been used for large-scale studies. we evaluated two protein-coding sequences as reporter genes for the yeast two-hybrid system, to determine if it was suitable as an alternative screening strategy. aspergillus awamori glucoamylase activity results in clear haloes around colonies producing this enzyme after growth on starch plates and staining with iodine vapors. however, transcription activatio ...200312917809
biochemical characterization of aspergillus awamori exoinulinase: substrate binding characteristics and regioselectivity of hydrolysis.1h-nmr analysis was applied to investigate the hydrolytic activity of aspergillus awamori inulinase. the obtained nmr signals and deduced metabolite pattern revealed that the enzyme cleaves off only fructose from inulin and does not possess transglycosylating activity. kinetics for the enzyme hydrolysis of inulooligosaccharides with different degree of polymerization (d.p.) were recorded. the enzyme hydrolyzed both beta2,1- as well as beta2,6-fructosyl linkages in fructooligosaccharides. from th ...200312922166
efficient production of arthromyces ramosus peroxidase by aspergillus awamori.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 produced. in contrast to several other studies about heterologous production of heme containing peroxidases, our results suggest that in a. awamori no h ...200312814876
expression of a synthetic copy of the bovine chymosin gene in aspergillus awamori from constitutive and ph-regulated promoters and secretion using two different pre-pro sequences.a copy of the bovine chymosin gene (chy) with a codon usage optimized for its expression in aspergillus awamori was constructed starting from synthetic oligonucleotides. to study the ability of this filamentous fungus to secrete bovine prochymosin, two plasmids were constructed in which the transcriptional, translational, and secretory control regions of the a. nidulans gpda gene and pepb genes were coupled to either preprochymosin or prochymosin genes. secretion of a protein enzymatically and i ...200312783481
improvement of foreign-protein production in aspergillus niger var. awamori by constitutive induction of the unfolded-protein response.unfolded-protein response (upr) denotes the upregulation of endoplasmic reticulum (er)-resident chaperone and foldase genes and numerous other genes involved in secretory functions during the accumulation of unfolded proteins into the er. overexpression of individual foldases and chaperones has been used in attempts to improve protein production in different production systems. we describe here a novel strategy to improve foreign-protein production. we show that the constitutive induction of the ...200314660339
role of two alpha-l-arabinofuranosidases in arabinoxylan degradation and characteristics of the encoding genes from shochu koji molds, aspergillus kawachii and aspergillus awamori.two different alpha-l-arabinofuranosidases from aspergillus kawachii were purified and characterized. the two enzymes acted synergically with xylanase in the degradation of arabinoxylan and resulted in an increase in the amount of ferulic acid release by feruloyl esterase. both enzymes were acidophilic and acid stable enzymes which had an optimum ph of 4.0 and were stable at ph 3.0-7.0. the general properties of the enzymes including ph optima and ph stability were similar to those of aspergillu ...200316233515
molecular cloning and characterization of an exoinulinase gene from aspergillus niger strain 12 and its expression in pichia pastoris.a genomic dna segment and cdnas encoding an extracellular exoinulinase from aspergillus niger strain 12 were cloned and sequenced. southern blot analysis indicated that the exoinulinase gene (inue) was present as a single copy in the genome. an open reading frame of 1611 by was interrupted by a single intron of 60 bp, and encoded a 19-amino acid signal peptide and a 518-amino acid mature protein. the mature protein contained a single cys residue and nine potential n-linked glycosylation sites. t ...200316233531
phenolic composition and radical scavenging activity of sweetpotato-derived shochu distillery by-products treated with koji.phenolic composition and radical scavenging activity in the shochu distillery by-products of sweetpotato (ipomoea batatas l.) treated with koji (aspergillus awamori mut.) and cellulase (cellulosin t2) were investigated to develop new uses. koji and cellulosin t2 treatment of shochu distillery by-products from sweetpotatoes, rice, and barley increased phenolic content. caffeic acid was identified as a dominant phenolic component in the shochu distillery by-products of the sweetpotato. adding koji ...200415618617
degradation of raw starch granules by alpha-amylase purified from culture of aspergillus awamori kt-11.raw-starch-digesting alpha-amylase (amyl iii) was purified to an electrophoretically pure state from the extract of a koji culture of aspergillus awamori kt-11 using wheat bran in the medium. the purified amyl iii digested not only soluble starch but also raw corn starch. the major products from the raw starch using amyl iii were maltotriose and maltose, although a small amount of glucose was produced. amyl iii acted on all raw starch granules that it has been tested on. however, it was consider ...200415469729
molecular cloning and determination of the nucleotide sequence of raw starch digesting alpha-amylase from aspergillus awamori kt-11.complementary dnas encoding alpha-amylases (amyl i, amyl iii) and glucoamylase (ga i) were cloned from aspergillus awamori kt-11 and their nucleotide sequences were determined. the sequence of amyl iii that was a raw starch digesting alpha-amylase was found to consist of a 1,902 bp open reading frame encoding 634 amino acids. the signal peptide of the enzyme was composed of 21 amino acids. on the other hand, the sequence of amyl i, which cannot act on raw starch, consisted of a 1,500 bp orf enco ...200415469730
crystal structure of exo-inulinase from aspergillus awamori: the enzyme fold and structural determinants of substrate recognition.exo-inulinases hydrolyze terminal, non-reducing 2,1-linked and 2,6-linked beta-d-fructofuranose residues in inulin, levan and sucrose releasing beta-d-fructose. we present the x-ray structure at 1.55a resolution of exo-inulinase from aspergillus awamori, a member of glycoside hydrolase family 32, solved by single isomorphous replacement with the anomalous scattering method using the heavy-atom sites derived from a quick cryo-soaking technique. the tertiary structure of this enzyme folds into two ...200415522299
[analysis of the ca2+ response of mycelial fungi to external effects by the recombinant aequorin method].using the mutant strain aspergillus awamori 66a producing a recombinant ca2+-dependent photosensitive protein aequorin, the dynamics of ca2+ was studied for the first time in the cytosol of the micromycetes exposed to stressful factors, such as an increase in extracellular ca2+ to 50 mm, hypoosmotic shock, and mechanical shock. cell response to stress proved to involve an increase in the ca2+ concentration in the cytosol, which was determined from the amplitude of aequorin luminescence and the t ...200415688931
proliferation of lactobacillus plantarum in solid-state fermentation of oats.in an attempt to introduce probiotic functionalities to breakfast cereals and similar food products, the technique of solid-state fermentation (ssf) was applied to cultivate lactobacillus plantarum (ncimb 8826) on oat bran and spent oats after lipid extraction by supercritical co(2) extraction. when compared to the frequently favored submerged processes for bacterium incubation, ssf presents not only the potential of simple downstream processing but also a more natural growth environment for the ...200414763831
thr/ser-rich domain of aspergillus glucoamylase is essential for secretion.the recombinant aspergillus awamori strain carrying the mutant glucoamylase-encoding gene in which the entire thr/ser-rich gp-i domain was deleted abolished secretion of mutant glucoamylase. the transcription of the bip-encoding bipa was low in the wild type (wt) strain, but elevated in the recombinant strain under the condition of glaa expression. the results indicate that the gp-i domain is vital for glucoamylase secretion.200415118335
calcium measurement in living filamentous fungi expressing codon-optimized aequorin.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 aspergillus awamori by codon optimization of the aequorin gene. three external stimuli (mechanical perturbation, hypo-osmotic shock and high external calcium) wer ...200415165245
modulation of aspergillus awamori thaumatin secretion by modification of bipa gene expression.two different strains, aspergillus awamori tgdth-4 and a. awamori tgp-3 overexpressing a synthetic gene encoding the plant sweet protein thaumatin, showed an unfolded protein response. to facilitate protein secretion, the chaperone bipa gene was expressed in a. awamori under control of the strong constitutive promoter of the gpda gene. a good correlation was observed between the level of the bipa transcript in different strains and the amount of thaumatin secreted. thaumatin secretion was increa ...200415345393
survey of mycoflora and ochratoxin a in dried vine fruits from argentina markets.the aims of this work were to identify the mycoflora and to evaluate the natural occurrence of oa in dried vine fruits. likewise, the capacity to produce oa by aspergillus section nigri was studied.200415355533
[effect of a chemical analogue of autoinducers of microbial anabiosis on the ca2+ response of mycelial fungi].the microbial alkylhydroxybenzenes (ahb), autoinducers of anabiosis, or d1 factors, participate in stress response of mycelial fungi, as determined from changes in intracellular ca2+ concentration. by using the genetically modified strain aspergillus awamori 66a, which produces a recombinant ca2+-dependent protein aequorin, the dynamics of ca2+ was studied in the cytosol of cells exposed to mechanical shock in the presence of the protective doses (0.001-0.01% w/vol) of a chemical ahb analogue, 4 ...200415688932
characterization of secretory genes ypt1/ypta and nsf1/nsfa from two filamentous fungi: induction of secretory pathway genes of trichoderma reesei under secretion stress conditions.two genes involved in protein secretion, encoding the rab protein ypt1/ypta and the general fusion factor nsfi/nsfa, were characterized from two filamentous fungi, trichoderma reesei and aspergillus niger var. awamori. the isolated genes showed a high level of conservation with their saccharomyces cerevisiae and mammalian counterparts, and t. reesei ypt1 was shown to complement yeast ypt1p depletion. the transcriptional regulation of the t. reesei ypt1, nsf1, and sar1 genes, involved in protein ...200414711675
a one-step method to convert vectors into binary vectors suited for agrobacterium-mediated transformation.bacterial artificial chromosomes (bacs) are widely used for the construction of physical maps, positional-cloning and whole-genome sequencing strategies. unfortunately, their use for functional genomics is limited, as currently there is no efficient method to use bacs directly for complementation. we describe a novel strategy for one-step conversion of any bac into a binary bac (bibac). using agrobacterium tumefaciens, these bibacs can be efficiently transformed to virtually all organisms, inclu ...200414745506
agrobacterium-mediated transformation of aspergillus awamori in the absence of full-length vird2, virc2, or vire2 leads to insertion of aberrant t-dna structures.reductions to 2, 5, and 42% of the wild-type transformation efficiency were found when agrobacterium mutants carrying transposon insertions in vird2, virc2, and vire2, respectively, were used to transform aspergillus awamori. the structures of the t-dnas integrated into the host genome by these mutants were analyzed by southern and sequence analyses. the t-dnas of transformants obtained with the vire2 mutant had left-border truncations, whereas those obtained with the vird2 mutant had truncated ...200415028687
the aspergillus nidulans amds gene as a marker for the identification of multicopy t-dna integration events in agrobacterium-mediated transformation of aspergillus awamori.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 with the transformation frequency obtained after hygromycin selection alone. however, it was found that 47% of the transformants obtained after hygromyci ...200415045526
role of bacterial virulence proteins in agrobacterium-mediated transformation of aspergillus awamori.the agrobacterium-mediated transformation of aspergillus awamori was optimized using defined co-cultivation conditions, which resulted in a reproducible and efficient transformation system. optimal co-cultivation conditions were used to study the role of agrobacterium tumefaciens virulence proteins in t-dna transfer. this study revealed that inactivation of either of the regulatory proteins (vira, virg), any of the transport pore proteins (virb), proteins involved in generation of the t-strand ( ...200415050546
restructuring upstream bioprocessing: technological and economical aspects for production of a generic microbial feedstock from wheat.restructuring and optimization of the conventional fermentation industry for fuel and chemical production is necessary to replace petrochemical production routes. guided by this concept, a novel biorefinery process has been developed as an alternative to conventional upstream processing routes, leading to the production of a generic fermentation feedstock from wheat. the robustness of aspergillus awamori as enzyme producer is exploited in a continuous fungal fermentation on whole wheat flour. vi ...200414760693
enzymatic production of caffeic acid by koji from plant resources containing caffeoylquinic acid derivatives.the effect of a koji (aspergillus awamori mut.) extract on the caffeoylquinic acid derivatives purified from sweetpotato (ipomoea batatas l.) leaves was examined to develop the mass production of caffeic acid. a koji extract hydrolyzed the caffeoylquinic acid derivatives, chlorogenic acid, 3,4-di-o-caffeoylquinic acid, 3,5-di-o-caffeoylquinic acid, 4,5-di-o-caffeoylquinic acid and 3,4,5-tri-o-caffeoylquinic acid, to caffeic acid. furthermore, the koji extract also converted the major polyphenoli ...200516195601
characteristics of transxylosylation by beta-xylosidase from aspergillus awamori k4.the aspergillus awamori k4 beta-xylosidase gene (xaw1) sequence was deduced by sequencing rt-pcr and pcr products. the orf was 2,412 bp and the predicted peptide was 804 amino acids long, corresponding to a molecular weight of 87,156 da. the mature protein was 778 amino acids long with a molecular weight of 84,632 da. a homology search of the amino acid sequence revealed that it was very similar to the aspergillus niger beta-xylosidase gene with only five amino acid differences. k4 beta-xylosida ...200516202538
agrobacterium-mediated transformation leads to improved gene replacement efficiency in aspergillus awamori.in this study, the efficiency of gene replacement in aspergillus awamori between agrobacterium-mediated transformation and cacl(2)/peg-mediated transformation was compared. for the genes, pyrg and gfaa, it was found that the homologous recombination frequencies obtained by agrobacterium-mediated transformation were 3- to 6-fold higher than the frequencies obtained with cacl(2)/peg protoplast transformation. for the pyrg gene, it was found that agrobacterium-mediated transformation allowed an eff ...200515588992
a new short-term toxicity assay using aspergillus awamori with recombinant aequorin gene.most currently available short-term toxicity assays are based on bacterial cells. therefore there is a need for novel eukaryotic microbial bioassays that will be relevant to higher eukaryotes such as animals and plants. ca2+ is a universal intracellular signalling molecule found in all organisms from prokaryotes to highly specialized animal cells. in fungi calcium has been demonstrated to be involved in control of many important processes. the recombinant aequorin gene from the jellyfish aequore ...200515992407
production of bifunctional proteins by aspergillus awamori: llama variable heavy chain antibody fragment (v(hh)) r9 coupled to arthromyces ramosus peroxidase (arp).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 v(hh) r9, resulting in the fusion expression cassettes arp-r9 or r9-arp. aspergillus awamori transformants were obtained which produced up to 30 mgl(-1) fusion protein in the culture medium. both fusion proteins showed peroxidase activity in an abts activity test. considerable amounts of fusion protein were detected intracellu ...200516169108
computer-aided analysis of spatial structure of some hydrolytic enzymes.using the molscript version 2.1 computer program for protein molecule modeling and x-ray structure analysis data the spatial structures of several hydrolytic enzymes have been compared. these include glucoamylase from aspergillus awamori and saccharomycopsis fibuligera and lipases from rhizopus japonicus. results on homology of amino acid sequences and topology of secondary structure elements were obtained. 3d models of these enzymes with positioning of functionally important groups in the activ ...200516271023
protein o-mannosyltransferase a of aspergillus awamori is involved in o-mannosylation of glucoamylase i.industrially important extracellular enzymes from filamentous fungi are often o-mannosylated. the structure and function of the pmta (aapmta) gene encoding the protein o-d-mannosyltransferase of aspergillus awamori were characterized. the aapmta disruptant, designated aapmta, was constructed by homologous recombination. the strain aapmta exhibited fragile cell morphology with respect to hyphal extension, as well as swollen hyphae formation and conidia formation in potato dextrose medium. moreove ...200516272387
free energy perturbation calculations on glucosidase-inhibitor complexes.free energy perturbation studies have been performed on glucoamylase ii (471) from aspergillus awamori var. x100 complexed with three different inhibitors: (+)lentiginosine, (+)(1s,2s,7r,8as) 1,2,7-trihydroxyindolizidine, (+)(1s,2s,7s,8as) 1,2,7-trihydroxyindolizidine and the inactive compound (+)(1s,7r,8as)-1,7-dihydroxyindolizidine. molecular dynamic simulations were carried out using a recently developed procedure for fast free energy perturbation calculations. in this procedure only a sphere ...200516787329
survey of coastal mangrove fungi for xylanase production and optimized culture and assay conditions.xylanase activity was detected among 34 of 77 fungal isolates derived from decaying wood, debris and soil samples collected in coastal mangrove environment of hong kong. of those, three isolates cy2809 (staganospora sp.), cy4786 and cy5040 were chosen for comparison of xylanase production in parallel to hu5048 (aspergillus awamori), a terrestrial, highly productive isolate. based on the assessment of mycelial biomass, xylanase activity and content of xylose-equivalent reducing sugars in their li ...200515847171
biochemical characterization of recombinant acetyl xylan esterase from aspergillus awamori expressed in pichia pastoris: mutational analysis of catalytic residues.we engineered an acetyl xylan esterase (awaxea) gene from aspergillus awamori into a heterologous expression system in pichia pastoris. purified recombinant awaxea (rawaxea) displayed the greatest hydrolytic activity toward alpha-naphthylacetate (c2), lower activity toward alpha-naphthylpropionate (c3) and no detectable activity toward acyl-chain substrates containing four or more carbon atoms. putative catalytic residues, ser(119), ser(146), asp(168) and asp(202), were substituted for alanine b ...200515848131
expression and production of llama variable heavy-chain antibody fragments (v(hh)s) by aspergillus awamori.we report the expression and production of llama variable heavy-chain antibody fragments (v(hh)s) by aspergillus awamori. fragments encoding v(hh)s were cloned in a suitable aspergillus expression vector and transformants secreting v(hh) fragments were analysed for integrated gene copy-numbers, mrna levels and protein production. functional v(hh)s were detected in the culture medium, indicating the feasibility of producing this type of protein in a fungal expression system. secreted v(hh)s were ...200515378291
mutational analysis of a feruloyl esterase from aspergillus awamori involved in substrate discrimination and ph dependence.we cloned the feruloyl esterase a gene from aspergillus awamori (awfaea) and engineered it to study substrate specificity and ph dependence of catalysis. based on the crystal structures of two type-a feruloyl esterases (fae-iii and anfaea) from aspergillus niger, residues located in the flap region of awfaea (asp71, thr72, asp77, and tyr80) were replaced with corresponding amino acid residues (ile, arg, asn, and phe), respectively, found in the lid of lipases from rhizomucor miehei (rmlip) and h ...200515716038
allosteric nadp-glutamate dehydrogenase from aspergilli: purification, characterization and implications for metabolic regulation at the carbon-nitrogen interface.nadp-dependent glutamate dehydrogenase (nadp-gdh) mediates fungal ammonium assimilation through reductive synthesis of glutamate from 2-oxoglutarate. by virtue of its position at the interface of carbon and nitrogen metabolism, biosynthetic nadp-gdh is a potential candidate for metabolic control. in order to facilitate characterization, a new and effective dye-affinity method was devised to purify nadp-gdh from two aspergilli, aspergillus niger and aspergillus nidulans. the a. niger nadp-gdh was ...200515870451
a novel system of genetic transformation allows multiple integrations of a desired gene in saccharomyces cerevisiae chromosomes.increasing industrial competitiveness and productivity demand that recombinant yeast strains, used in many different processes, be constantly adapted and/or genetically improved to suit changing requirements. among yeasts, saccharomyces cerevisiae is the best-studied organism, and the most frequently employed yeast in industrial processes. in the present study, laboratory strains and industrial s. cerevisiae strains were stably transformed with a novel vector containing the glucoamylase cdna of ...200616831478
[the effect of acid proteinases on the activity and stability of glucoamylase preparations].it was demonstrated that the presence of acid proteinases in the preparation isolated from aspergillus awamori decreased the activity and stability of glucoamylase. patterns of changes in the enzymatic activity and stability of glucoamylase at increased temperature and various ph values were studied over a long-term storage. a biospecific sorbent for removal of acid proteinases was synthesized, and glucoamylase preparations free of proteolytic activity were produced.200616761575
enhanced antioxidative activity of soybean koji prepared with various filamentous fungi.in the present study, soybean koji fermented with various gras filamentous fungi, including aspergillus sojae bcrc 30103, aspergillus oryzae bcrc 30222, aspergillus awamori, actinomucor taiwanensis and rhizopus sp. these organisms are commonly used as starters in the fermentation of many traditional, oriental food products. the growth of starter organisms, total phenolic content, and antioxidative activities of the methanol extract of these kojis are compared with specific reference to alpha-dip ...200616943061
suppression on the mutagenicity of 4-nitroquinoline-n-oxide by the methanol extracts of soybean koji prepared with various filamentous fungi.in this study, solid fermentation of soybean with various gras filamentous fungi including aspergillus sojae bcrc 30103, aspergillus oryzae bcrc 30222, aspergillus awamori, actinomucor taiwanesis and rhizopus sp. was performed to prepare various soybean kojis. toxicity, mutagenicity and suppression on the mutagenesis induced by a direct mutagen, 4-nitroquinoline-n-oxide (4-nqo) on salmonella typhimurium ta 100, by the various methanol extracts of the prepared soybean koji and unfermented soybean ...200616580082
digestion of dna regions to discriminate ochratoxigenic and non-ochratoxigenic strains in the aspergillus niger aggregate.aspergillus strains belonging to the aspergillus niger aggregate, either isolated from italian grapes or received from public collections, were analysed in order to discriminate between the ochratoxin a (ota) producing and the non-producing strains by means of the analysis of internal transcribed spacers (its), intergenic spacers (igs) and of a beta-tubulin gene portion. a. niger and aspergillus awamori were identified observing the macro- and microscopic features of the colonies and the strains ...200616735072
n-linked oligosaccharides of aspergillus awamori feruloyl esterase are important for thermostability and catalysis.a unique n-linked glycosylation motif (asn(79)-tyr-thr) was found in the sequence of type-a feruloyl esterases from aspergillus spp. to clarify the function of the flap, the role of n-linked oligosaccharides located in the flap region on the biochemical properties of feruloyl esterase (awfaea) from aspergillus awamori expressed in pichia pastoris was analyzed by removing the n-linked glycosylation recognition site by site-directed mutagenesis. n79 was replaced with a or q. n-glycosylation-free n ...200617031032
enzymatic degradation of oligosaccharides in pinto bean flour.the use of dry edible beans is limited due to the presence of flatulence factors, the raffinose oligosaccharides. our objective was to investigate the process for the removal of oligosaccharides from pinto bean using enzymatic treatment and to compare it to removal by soaking and cooking methods. crude enzyme preparation was produced by six fungal species on wheat bran- and okara-based substrates with soy tofu whey. the loss of raffinose oligosaccharides after soaking pinto beans for 16 h at the ...200616478251
distribution profiles of isoflavone isomers in black bean kojis prepared with various filamentous fungi.this study was conducted to compare the transformation of both isoflavone derivatives (aglycones, beta-glucosides, and acetyl and malonyl gluclucosides) and beta-glucosidase activity in kojis fermented with various generally recognized as safe filamentous fungi including aspergillus awamori, aspergillus oryzae, aspergillus sojae, rhizopus azygosporus, and rhizopus sp. no. 2. solid fermentation was performed to prepare the kojis by inoculating the steamed black beans with starter organism and cul ...200616478253
evidence of rip (repeat-induced point mutation) in transposase sequences of aspergillus oryzae.a dna methyl-binding column was used to isolate genomic fragments enriched for dna-methylation from aspergillus parasiticus. one of the isolated sequences presented 67% identity at the protein level with the transposase from the transposable element tan1 of aspergillus niger var. awamori, and was found to be present in at least 20 copies in the aspergillus oryzae database. analysis of four copies showed evidence of c:g to t:a transitions in at least 98.2% of the mutations found over a 1,032-1,18 ...200616531081
rock phosphate enriched compost: an approach to improve low-grade indian rock phosphate.in this study, rock phosphate enriched composts (rp-compost) were prepared by mixing four low-grade indian rock phosphates with rice straw with and without aspergillus awamori. rp-compost had higher total p, citrate soluble p (csp), organic p (org.p), acid and alkaline phosphatase activities, and lower water soluble p (wsp) and microbial biomass c (mbc) than normal compost. inoculation with a. awamori increased total p, wsp, csp, org.p, mbc and acid phosphatase activity. rp-compost recorded lowe ...200616545954
microbial biomass, p-nutrition, and enzymatic activities of wheat soil in response to phosphorus enriched organic and inorganic manures.compost was prepared from wheat straw enriched with rajasthan rock phosphate and aspergillus awamori. the resulting phospho-compost along with phosphorus enriched fym, mineral fertilizer (rock phosphate) and super phosphate were evaluated for their individual contribution in improving organic matter status, p availability, and enzymatic activities of soil under wheat crop grown in a micro plot. the results showed that total organic carbon, nitrogen, microbial biomass, and humus content (an index ...200616578904
purification and physico-chemical characterisation of genetically modified phytases expressed in aspergillus awamori.two heterologous phytases from aspergillus awamori and aspergillus fumigatus obtained from submerged cultures of genetically modified fungal strains in addition to two commercially available phytase preparations (allzyme and natuphos phytases) were purified to homogeneity using a combination of ultrafiltration, gel filtration and ion exchange. the purified preparations were used in subsequent characterisation studies, in which western immunoblot analysis, ph and temperature optima, thermal stabi ...200616243522
biochemical and kinetic analysis of the gh3 family beta-xylosidase from aspergillus awamori x-100.the beta-xylosidase from aspergillus awamori x-100 belonging to the family 3 glycoside hydrolase revealed a distinctive transglycosylating ability to produce xylooligosaccharides with degree of polymerization more than 7. in order to explain this fact, the enzyme has been subjected to the detailed biochemical study. the enzymatic hydrolysis of p-nitrophenyl beta-d-xylopyranoside was found to occur with overall retention of substrate anomeric configuration suggesting cleavage of xylosidic bonds t ...200717145041
cereal-based biorefinery development: integrated enzyme production for cereal flour hydrolysis.restructuring the traditional fermentation industry into viable biorefineries for the production of fuels, chemicals and plastics is essential in order to replace (petro)chemical processing. this work presents engineering aspects of aspergillus awamori submerged fermentation for on-site production of an enzymatic consortium that contains glucoamylase, protease and phosphatase. the crude broth filtrate was used for the production of wheat flour hydrolysates. improvements on traditional starch hyd ...200717009318
apho1 from the yeast arxula adeninivorans encodes an acid phosphatase of broad substrate specificity.the extracellular acid phosphatase-encoding arxula adeninivorans apho1 gene was isolated using degenerated specific oligonucleotide primers in a pcr screening approach. the gene harbours an orf of 1449 bp encoding a protein of 483 amino acids with a calculated molecular mass of 52.4 kda. the sequence includes an n-terminal secretion sequence of 17 amino acids. the deduced amino acid sequence exhibits 54% identity to phytases from aspergillus awamori, asp. niger and asp. ficuum and a more distant ...200717016743
enhancement of acid amylase production by an isolated aspergillus awamori.evaluation of the influence of fermentation components on extracellular acid amylase production by an isolated fungal strain aspergillus awamori.200717184336
application of statistical experimental design to optimize culture requirements of aspergillus sp. zh-26 producing xylanase for degradation of arabinoxylans in mashing.the objective of this study was to isolate and characterize a strain of aspergillus capable of producing xylanase. according to the morphology and comparison of its (internal transcribed spacer) rdna gene sequence, the strain aspergillus sp. zh-26 was identified as a strain of aspergillus awamori. statistically based experimental designs were applied for the optimization of xylanase production from a. awamori zh-26. the considered medium components included 17 components as follows: yeast extrac ...200717995733
mutagenic and antimutagenic effects of methanol extracts of unfermented and fermented black soybeans.in this study, solid fermentation of steamed black soybean with various gras (generally recognized as safe) filamentious-fungi including aspergillus awamori, aspergillus oryzae bcrc 30222, aspergillus sojae bcrc 30103, rhizopus azygosporus bcrc 31158 and rhizopus sp. no. 2 was performed. mutagenicity and antimutagenicity of the methanol extracts of unfermented and fermented steamed black soybeans against 4-nitroquinoline-n-oxide (4-nqo), a direct mutagen and benzo[a]pyrene (b[a]p), an indirect m ...200717628128
kinetics of phosphorus and potassium release from rock phosphate and waste mica enriched compost and their effect on yield and nutrient uptake by wheat (triticum aestivum).an attempt was made to study the efficient use of rice straw and indigenous source of phosphorus and potassium in crop production through composting technology. various enriched composts were prepared using rice straw, rock phosphate (rp), waste mica and bioinoculant (aspergillus awamori) and kinetics of release of phosphorus and potassium from enriched composts and their effect on yield and nutrient uptake by wheat (triticum aestivum) were carried out. results showed sharp increases in release ...200817905580
heterologous expression of two ferulic acid esterases from penicillium funiculosum.two recombinant ferulic acid esterases from penicillium funiculosum produced in aspergillus awamori were evaluated for their ability to improve the digestibility of pretreated corn stover. the genes, faea and faeb, were cloned from p. funiculosum and expressed in a. awamori using their native signal sequences. both enzymes contain a catalytic domain connected to a family 1 carbohydrate-binding module by a threonine-rich linker peptide. interestingly, the carbohydrate binding-module is n-terminal ...200818421589
agrobacterium-mediated transformation of the filamentous fungus aspergillus awamori.many transformation methods have been developed to introduce dna into filamentous fungi. one of these methods is agrobacterium-mediated transformation (amt). here, we describe an efficient protocol for amt of aspergillus awamori. this protocol has been used to determine the function of agrobacterium virulence genes during amt, to identify factors influencing transformation frequencies, to generate insertional mutants and to generate a. awamori gene knockout transformants. this protocol in not on ...200818833205
a wheat biorefining strategy based on solid-state fermentation for fermentative production of succinic acid.in this study, a novel generic feedstock production strategy based on solid-state fermentation (ssf) has been developed and applied to the fermentative production of succinic acid. wheat was fractionated into bran, gluten and gluten-free flour by milling and gluten extraction processes. the bran, which would normally be a waste product of the wheat milling industry, was used to produce glucoamylase and protease enzymes via ssf using aspergillus awamori and aspergillus oryzae, respectively. the r ...200818434138
implications of cellobiohydrolase glycosylation for use in biomass conversion.the cellulase producing ascomycete, trichoderma reesei (hypocrea jecorina), is known to secrete a range of enzymes important for ethanol production from lignocellulosic biomass. it is also widely used for the commercial scale production of industrial enzymes because of its ability to produce high titers of heterologous proteins. during the secretion process, a number of post-translational events can occur, however, that impact protein function and stability. another ascomycete, aspergillus niger ...200818471276
recombinant human lactoferrin and iron transport across caco-2 monolayers: effect of heat treatment on the binding to cells.recombinant human lactoferrin (rhlf) from aspergillus awamori bound to caco-2 cell membranes in a saturable manner. the dissociation constant for the apo form was (kd)=2.2 x 10(-7) m; however, the specific binding of the iron-saturated rhlf and of lactoferrin from human milk (hlf) was too low to calculate the binding parameters. recombinant human lactoferrin subjected to heat treatment did not lose the ability to bind to cell membranes except at high temperature and long time treatments (85 and ...200818355020
udp-xylose-stimulated glucuronyltransferase activity in wheat microsomal membranes: characterization and role in glucurono(arabino)xylan biosynthesis.microsomal membranes from etiolated wheat (triticum aestivum) seedlings cooperatively incorporated xylose (xyl), arabinose, and glucuronic acid residues from their corresponding uridine 5'-diphosphosugars into an ethanol-insoluble glucurono(arabino)xylan (gax)-like product. a glucuronyltransferase activity that is enhanced by the presence of udp-xyl was also identified in these microsomes. wheat glucuronyltransferase activity was optimal at ph 7 and required manganese ions, and several lines of ...200818359844
heterologous expression of aspergillus niger beta-d-xylosidase (xlnd): characterization on lignocellulosic substrates.the gene encoding a glycosyl hydrolase family 3 xylan 1,4-beta-xylosidase, xlnd, was successfully cloned from aspergillus niger strain atcc 10864. the recombinant product was expressed in aspergillus awamori, purified by column chromatography, and verified by matrix-assisted laser desorption ionization, tandem time of flight (maldi-tof/tof) mass spectroscopy of tryptic digests. the t (max) was determined using differential scanning microcalorimetry (dsc) to be 78.2 degrees c; the k (m) and k (ca ...200818421587
characteristics of glucose uptake by glucose- and nh4-limited grown penicillium ochrochloron at low, medium and high glucose concentration.glucose uptake by penicillium ochrochloron (formerly penicillium simplicissimum) was studied from 0.01 to 400 mm glucose using chemostat culture and bioreactor batch culture. the characteristics of glucose uptake varied considerably with the conditions of growth, harvest and uptake assay. glucose-limited grown mycelium showed one saturable transport system [k(s) below 0.01 mm; v(max) 1.1-1.2 mmol (g dry weight)(-1)h(-1)] plus a first order process (permeability p=1.2x10(-7)cm s(-1)). ammonium-li ...200818722543
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