a new transcriptional activator for amylase genes in aspergillus.we have cloned a regulatory gene for amylase synthesis in aspergillus oryzae. this gene, amyr, encodes a 604-amino acid transcriptional activator with a cys6 zinc cluster, that shows extensive homology to the dna binding domain of gal4 from saccharomyces cerevisiae. the dna binding domain of amyr binds to two types of sequences found in a number of promoters from aspergillus genes coding for starch-degrading enzymes. one type of binding site is characterized by two cgg triplets separated by eigh ...199910628849
cloning and characterization of a gene (arpa) from aspergillus oryzae encoding an actin-related protein required for normal nuclear distribution and morphology of conidiophores.we have isolated the arpa gene from aspergillus oryzae as a homologue of the neurospora crassa ro-4 gene. in n. crassa, mutations in the ro-4 gene, which encodes a major component of the dynactin complex arp1, causes curling of hyphae and abnormalities in nuclear distribution. the arpa gene contains two introns and encodes a polypeptide of 381 amino acids, with a 78% sequence identity to the n. crassa arp1. overexpression of the arpa gene causes a defect in nuclear migration into elongating hyph ...199910628858
biochemical analysis of recombinant fungal mutanases. a new family of alpha1,3-glucanases with novel carbohydrate-binding domains.nucleotide sequence analysis shows that trichoderma harzianum and penicillium purpurogenum alpha1,3-glucanases (mutanases) have homologous primary structures (53% amino acid sequence identity), and are composed of two distinct domains: a nh(2)-terminal catalytic domain and a putative cooh-terminal polysaccharide-binding domain separated by a o-glycosylated pro-ser-thr-rich linker peptide. each mutanase was expressed in aspergillus oryzae host under the transcriptional control of a strong alpha-a ...200010636904
myelopoietic response in tumour-bearing mice by an aggregated polymer isolated from aspergillus oryzae.the effects of magnesium ammonium phospholinoleate-palmitoleate anhydride (mapa), a proteic aggregated polymer isolated from aspergillus oryzae, on the growth and differentiation of granulocyte-macrophage progenitor cells (colony-forming unit-granulocyte-macrophage [cfu-gm]) in normal and ehrlich ascites tumour-bearing mice were studied. myelosuppression concomitant with increased numbers of spleen cfu-gm was observed in tumour-bearing mice. treatment of these animals with mapa (0.5-10 mg/kg) st ...200010675729
a spectrophotometric method for assay of tannase using rhodanine.a method for assay of microbial tannase (tannin acyl hydrolase) based on the formation of chromogen between gallic acid and rhodanine is reported. unlike the previous protocols, this method is sensitive up to gallic acid concentration of 5 nmol and has a precision of 1.7% (relative standard deviation). the assay is complete in a short time, very convenient, and reproducible.200010683234
effect of temperature and enzyme origin on the enzymatic synthesis of oligosaccharides.the aim of this research is to quantify the effect of temperature and enzyme origin on the enzymatic synthesis of oligosaccharides. quantification of these effects is important because temperature and enzyme origin are important process parameters. a kinetic model was used to describe the concentrations in time. the kinetic parameters were determined by using data obtained in batch experiments at various temperatures (20, 30, 40, and 50 degrees c) and by using beta-galactosidases from bacillus c ...200010689088
reversible enzyme immobilization via a very strong and nondistorting ionic adsorption on support-polyethylenimine tailor-made anionic exchange resins have been prepared, based on films of large polyethylenimine polymers (e.g., mw 25,000) completely coating, via covalent immobilization, the surface of different porous supports (agarose, silica, polymeric resins). most proteins contained in crude extracts from different sources have been very strongly adsorbed on them. ionic exchange properties of such composites strongly depend on the size of polyethylenimine polymers as well as on the exact conditions o ...200010699877
is fungal alpha-amylase in bread an allergen?the enzyme alpha-amylase from aspergillus oryzae used in bakeries to improve the bread quality has been identified as an inhalative allergen in baker's asthma. it is doubtful whether this enzyme can induce allergic sensitization in regular bread consumers.200010718854
production of extracellular lipases by penicillium cyclopium purification and characterization of a partial acylglycerol lipase.penicillium cyclopium, grown in stationary culture, produces a type i lipase specific for triacylglycerols while, in shaken culture, it produces a type ii lipase only active on partial acylglycerols. lipase ii has been purified by ammonium sulfate precipitation and chromatographies on sephadex g-75 and deae-sephadex. the enzyme exists in several glycosylated forms of 40-43 kda, which can be converted to a single protein of 37 kda by enzymatic deglycosylation. activity of lipase ii is maximal at ...200010737172
a simple and rapid method for the preparation of a cell-free extract with ccaat-binding activity from filamentous fungi.a simple and rapid method for the preparation of a cell-free extract with the ccaat-binding activity was established with aspergillus nidulans as a model fungus. proteins were extracted with 6 m guanidine hydrochloride directly from mycelia and renatured by dialysis. this method was found applicable to other filamentous fungi such as aspergillus oryzae and trichoderma viride.200010737212
efficient heterologous expression in aspergillus oryzae of a unique dye-decolorizing peroxidase, dyp, of geotrichum candidum dec 1.efficient expression of the dye-decolorizing peroxidase, dyp, from geotrichum candidum dec 1 in aspergillus oryzae m-2-3 was achieved by fusing mature cdna encoding dyp with the a. oryzae alpha-amylase promoter (amyb). the activity yield of the purified recombinant dyp (rdyp) was 42-fold compared with that of the purified native dyp from dec 1. no exogenous heme was necessary for the expression of rdyp in a. oryzae. from the n-terminal amino acid sequence analyses of native dyp and rdyp, the abs ...200010742277
directed deletions in the aflatoxin biosynthesis gene homolog cluster of aspergillus investigate the structure of the aflatoxin gene cluster in aspergillus oryzae, 39 strains belonging to this species were examined for the existence of pksa, fas1a, aflr and vbs, and the results compared with those for ver-1 obtained previously. these five genes are involved in aflatoxin biosynthesis in aspergillus parasiticus. the strains examined were categorized into three groups; group 1, having the five homologs; 2, having ver-1 and vbs; and 3, having vbs homologs. long-pcr analysis of th ...200010743566
evidence that the glucoamylases and alpha-amylase secreted by aspergillus niger are proteolytically processed products of a precursor enzyme.a 125-kda starch hydrolysing enzyme of aspergillus niger characterised by its ability to dextrinise and saccharify starch [suresh et al. (1999) appl. microbiol. biotechnol. 51, 673-675] was also found to possess activity towards raw starch. segregation of these activities in the 71-kda glucoamylase and a 53-kda alpha-amylase-like enzyme supported by antibody cross-reactivity studies and the isolation of mutants based on assay screens for the secretion of particular enzyme forms revealed the 125- ...200010767433
the influence of nitrogen sources on the alpha-amylase productivity of aspergillus oryzae in continuous cultures.the influence of the nitrogen source on the alpha-amylase productivity of aspergillus oryzae was quantified in continuous cultivations. both inorganic and complex nitrogen sources were investigated and glucose was used as the carbon and energy sources. for production of alpha-amylase, nitrate was shown to be inferior to ammonia as a nitrogen source. a mixture of ammonia and complex nitrogen sources, such as yeast extract or casein hydrolysate, was better than with ammonia as the sole nitrogen so ...200010772466
a novel nuclear factor, sreb, binds to a cis-acting element, sre, required for inducible expression of the aspergillus oryzae taka-amylase a gene in a. nidulans.the taka-amylase a gene (taag2) of aspergillus oryzae is inducibly expressed in a. nidulans upon exposure to inducing carbon sources, such as starch and maltose. in order to identify nuclear factor(s) possibly involved in the induction of the taag2 gene, gel mobility shift assays and dnase i footprinting analyses were carried out, and revealed a novel nuclear factor in a. nidulans extracts, which specifically bound to two sites in the taag2 promoter region, -204 to -189 and -182 to -168, which s ...200010778741
using dna-tagged mutagenesis to improve heterologous protein production in aspergillus oryzae.using dna-tagged mutagenesis to improve heterologous protein production in aspergillus oryzae. fungal genetics and biology 29, 28-37. restriction enzyme-mediated integration (remi) has been employed as a mutagen to generate two insertion libraries in an aspergillus oryzae strain expressing a thermomyces lanuginosus lipase. the remi libraries were created using linearized plasmid containing the a. oryzae pyrg and either bamhi or ecori enzyme. the libraries were screened for lipase production, and ...200010779397
molecular cloning and characterization of rpba encoding rna polymerase ii largest subunit from a filamentous fungus, aspergillus oryzae.we have cloned rpba encoding the rna polymerase ii largest subunit (poliil) from a filamentous fungus, aspergillus oryzae. the rpba product included eight highly conserved regions and the carboxyl-terminal domain (ctd). a. oryzae poliil ctd with 184 amino acids was composed of 25 ctd consensus repeats, which was a similar number to those of lower eukaryotes. the amino acids in each repeat of a. oryzae poliil, however, conformed less to the ctd consensus than those of poliils from other lower euk ...200010803973
agitation induced mycelial fragmentation of aspergillus oryzae and penicillium chrysogenum.given the impact of mycelial morphology on fermentation performance, it is important to understand the factors that influence it, including agitation-induced fragmentation. the successful application of the energy dissipation/circulation function (edc) to correlate fragmentation of penicillium chrysogenum with agitation intensity and with different impeller types [5] has already been demonstrated. the edc function takes into account the specific energy dissipation rate in the impeller swept volu ...200010817815
unique catalytic and molecular properties of hydrolases from aspergillus used in japanese bioindustries.this review covers the unique catalytic and molecular properties of three proteolytic enzymes and a glycosidase from aspergillus. an aspartic proteinase from a. saitoi, aspergillopepsin i (ec, favors hydrophobic amino acids at p1 and p'1 like gastric pepsin. however, aspergillopepsin i accommodates a lys residue at p1, which leads to activation of trypsinogens like duodenum enteropeptidase. substitution of asp76 to ser or thr and deletion of ser78, corresponding to the mammalian aspar ...200010830477
galactosylation of thiol group by beta-galactosidase.beta-galactosidase catalyzed beta-galactosylation not only of a hydroxyl group but also of a thiol group in the condensation reaction of d-galactose and 2-mercaptoethanol. the thio-galactosylation product was confirmed as 2-hydroxyethyl s-beta-d-galactoside on the bases of fast atom bombardment mass spectrometry, infrared spectroscopy, and nuclear magnetic resonance spectorometry. aspergillus oryzae beta-galactosidase hydrolyzed p-nitrophenyl s-beta-d-galactoside most rapidly among several beta- ...200010830485
molecular cloning and characterization of a transcriptional activator gene, amyr, involved in the amylolytic gene expression in aspergillus oryzae.a gene, designated amyr, coding for a transcriptional activator involved in amylolytic gene expression has been cloned from aspergillus oryzae by screening for a clone that enabled to reverse the reduced expression of the alpha-amylase gene (amyb) promoter. amyr encodes 604 amino acid residues of a putative dna-binding protein carrying a zinc binuclear cluster motif (zn(ii)2cys6) belonging to the gal4 family of transcription factors. the amyr gene disruptants showed a significant restricted grow ...200010830498
comprehensive cloning and expression analysis of glycolytic genes from the filamentous fungus, aspergillus oryzae.we cloned all the glycolytic genes from aspergillus oryzae and analyzed their transcriptional regulation by the carbon source in the medium. the deduced amino-acid sequences of the glycolytic genes showed high identity (approximately 41-93%) to those from other lower eukaryotes. genomic southern hybridization indicated that all the genes existed as a single copy in the genome. comparison of mrna levels between mycelia grown on glucose and on pyruvate showed that most of the a. oryzae glycolytic ...200010853769
a novel approach to the recovery of biologically active oligosaccharides from milk using a combination of enzymatic treatment and nanofiltration.a new easily scalable approach to the recovery of biologically active oligosaccharides from milk has been developed which relies on the combination of enzymatic treatment of defatted milk using beta-galactosidase and nanofiltration. it was shown that enzymatic hydrolysis of lactose significantly improves the efficiency and selectivity of membrane-based separations. with the best membrane, as much as 6.7 g of oligosaccharides (containing very little contaminating lactose) could be obtained from o ...200010862685
immobilization/stabilization on eupergit c of the beta-galactosidase from b. circulans and an alpha-galactosidase from aspergillus oryzae.two synthetically useful glycosidases, the beta-galactosidase from bacillus circulans and an alpha-galactosidase from aspergillus oryzae have been immobilized on eupergit c. the immobilized enzymes retain high catalytic activity and show increased thermal stability compared with the free enzymes.200010862898
[comparative characteristics of proteolytic enzyme preparations by degree of hydrolysis of a microbial protein].proteolytic enzymatic preparations obtained from fungi and bacteria were compared by their ability to hydrolyze yeast protein. fungal preparations were more effective. there was a more than twofold increase in the level of amine nitrogen in cell biomass hydrolysates in of cell biomass comparison to that induced by bacterial preparations. the amino acid composition of these hydrolysates was studied. amyloprotooryzin, a preparation from aspergillus oryzae 387, displayed the highest potency in prof ...200010867948
enzymatic formation of ether linkage producing shoyuflavones from genistein and (+/-)-trans-epoxysuccinic acid.the production mechanism of shoyuflavones, conjugated ethers of isoflavones with tartaric acid and isolated from fermented soy sauce, was studied. in the high molecular weight fraction of the culture extract of aspergillus oryzae, genistein was transformed into shoyuflavone b in the presence of (+/-)-trans-epoxysuccinic acid but not in the low molecular one. asp. sojae and asp. tamarii showed high activity similar to asp. oryzae but none of asp. niger, rhizopus oligosporus, and mucor praini did. ...200010888513
induction of the soybean phytoalexins coumestrol and glyceollin by aspergillus.several isoflavonoid phytoalexins produced by soybeans are known to be estrogenic, with potential beneficial health effects in humans. increased production of phytoalexins by the soybean plant will facilitate research efforts in this area. in this study, phytoalexin induction and accumulation in soybean cotyledon tissue was observed using four species of aspergillus: a. sojae, a. oryzae, a. niger, and a. flavus. all four aspergillus species tested elicited phytoalexin accumulation in living soyb ...200010888516
noncompetitive, reversible inhibition of aminoacylase-1 by a series of l-alpha-hydroxyl and l-alpha-fluoro fatty acids: ligand specificity of aspergillus oryzae and porcine kidney enzymes.l-lactate and l-beta-phenyllactate have been identified in the culture broth of streptomyces sp. ky-11 as reversible noncompetitive inhibitors of aspergillus oryzae aminoacylase-1 and porcine kidney aminoacylase i. a series of alpha-hydroxyl acids (dl-r-ch(oh)-cooh, r = et, n-pro, n-butyl, n-pentyl, n-hexyl) also inhibited the two enzymes in reversible noncompetitive kinetics, and the inhibition potency (-log k(i)) increased with the increased hydrophobicity of the r group. the two eukaryotic en ...200010898943
a non-specific aminopeptidase from aspergillus.a fermentation broth supernatant of the aspergillus oryzae strain atcc20386 contains aminopeptidase activity that releases a wide variety of amino acids from natural peptides. the supernatant was fractionated by anion exchange chromatography. based on the primary amino acid sequence data obtained from proteins in certain fractions, polymerase chain reaction (pcr) primers were made and a pcr product was generated. this pcr product was used to screen an a. oryzae cdna library from which the full l ...200010899618
identification of functional elements that regulate the glucoamylase-encoding gene (glab) expressed in solid-state culture of aspergillus oryzae.aspergillus oryzae has-two glucoamylase-encoding genes, glaa and glab, whose expressions are distinguished by the type of culture used. the glab gene is markedly expressed in solid-state culture but is little expressed in submerged culture. in solid-state culture, glab expression at the transcriptional level is enhanced by low-aw (water activity), high-temperature, and physical barriers to hyphal extension, as well as by starch. to determine the cis-acting factors in the glab promoter, deletion ...200010905427
an aspergillus oryzae ccaat-binding protein, aocp, is involved in the high-level expression of the taka-amylase a gene.aspergillus oryzae contains a nuclear protein designated aocp, which binds specifically to a ccaat sequence in the promoter region of the a. oryzae taka-amylase a gene. a gene encoding a homologue of aspergillus nidulans hapc, a subunit of the a. nidulans ccaat binding complex, was isolated from a. oryzae and designated aohapc. aohapc comprises 215 amino acids and shows 84% identity to a. nidulans hapc. transformation of the a. nidulans hapc deletion strain with the aohapc gene restored the ccaa ...200010905428
enzymatic synthesis of 1,3,6,8-tetrahydroxynaphthalene solely from malonyl coenzyme a by a fungal iterative type i polyketide synthase pks1.the colletotrichum lagenarium pks1 gene encoding iterative type i polyketide synthase of 1,3,6,8-tetrahydroxynaphthalene (t4hn) was overexpressed in aspergillus oryzae. sds-page analysis of the cell-free extract prepared from the transformant showed an intense band of 230000 which corresponded to the molecular weight of the deduced pks1 protein. by using this cell-free extract, in vitro synthesis of t4hn was successfully confirmed as the first example of the fungal multi-aromatic ring polyketide ...200010913297
conversion of 1-benzoylindole by aspergillus strains.biotransformation of 1-benzoylindole (bi) by the strains aspergillus flavus vkm f-1024 and aspergillus oryzae vkm f-44 was studied. the major metabolites isolated were identified as 4-hydroxyindole (4-hi), 5-hydroxyindole (5-hi), 4-hydroxy- -benzoylindole, 4-hydroxy-1-(4'-hydroxy)-benzoylindole and indole. the structure of the metabolites was determined by mass spectrometry and proton nuclear magnetic resonance spectroscopy. the pathways of bi metabolism via initial monohydroxylation at c-4 and ...200010919329
cloning and sequencing of the triacylglycerol lipase gene of aspergillus oryzae and its expression in escherichia coli.aspergillus oryzae produces at least three extracellular lipolytic enzymes, l1, l2 and l3 (cutinase, mono- and diacylglycerol lipase, and triacylglycerol lipase, respectively). we cloned the triacylglycerol lipase gene (provisionally designated tgla) by screening a genomic library using a pcr product obtained with two degenerate oligonucleotide primers corresponding to amino acid sequences of l3 as probes. nucleotide sequencing of the genomic dna and cdna revealed that the l3 gene (tgla) has an ...200010930731
isolation of replicational cue elements from a library of bent dnas of aspergillus oryzae.two fragments that could function as replicational cue elements were isolated from a genomic dna digest of aspergillus oryzae on the basis of abnormal behavior in polyacrylamide gel electrophoresis. the vector used in this study contained a scaffold-associated region of the drosophila melanogaster ftz gene to provide nuclear retention. neither fragment contained a yeast ars consensus sequence or an eukaryotic topoisomerase ii binding sequence. one of the fragments showed sequence homology with t ...200010939521
pyrithiamine resistance gene (ptra) of aspergillus oryzae: cloning, characterization and application as a dominant selectable marker for transformation.a pyrithiamine (pt) resistance gene (ptra) was cloned from a genomic dna library prepared from a pt resistant mutant of aspergillus oryzae. it conferred high resistance to pt on an a. oryzae industrial strain as well as a. nidulans. nucleotide sequence analysis showed that the ptra gene contained one intron (58-bp) and encodes 327 amino acid (aa) residues. additionally, the deduced aa sequence has 72% and 63% identity to fusarium solani sti35 encoding a stress-inducible protein and saccharomyces ...200010945258
identification of copper ligands in aspergillus oryzae tyrosinase by site-directed mutagenesis.copper ligands of the recombinant tyrosinase from the fungus aspergillus oryzae expressed in saccharomyces cerevisiae or escherichia coli were identified by site-directed mutagenesis. the recombinant protyrosinases expressed in s. cerevisiae were assayed for catalytic activities of mono-oxygenase and l-dopa oxidase at ph 5.5 after acid shock at ph 3.0. replacements of his-63, his-84, his-93, his-290, his-294, his-332 or his-333 with asparagine resulted in mutant enzymes exhibiting no activities. ...200010947969
molecular cloning and characterization of a gene encoding glutaminase from aspergillus oryzae.a glutaminase from aspergillus oryzae was purified and its molecular weight was determined to be 82,091 by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. purified glutaminase catalysed the hydrolysis not only of l-glutamine but also of d-glutamine. both the molecular weight and the substrate specificity of this glutaminase were different from those reported previously [yano et al. (1998) j ferment technol 66: 137-143]. on the basis of its internal amino acid sequen ...200010952006
fungal morphology and fragmentation behavior in a fed-batch aspergillus oryzae fermentation at the production is well known that high-viscosity fermentation broth can lead to mixing and oxygen mass transfer limitations. the seemingly obvious solution for this problem is to increase agitation intensity. in some processes, this has been shown to damage mycelia, affect morphology, and decrease product expression. however, in other processes increased agitation shows no effect on productivity. while a number of studies discuss morphology and fragmentation at the laboratory and pilot scale, there are rela ...200010992234
hydrolysis of beta-galactosyl ester linkage by beta-galactosidases.p-hydroxybenzoyl beta-galactose (phb-gal) was synthesized chemically to examine the hydrolytic activity of beta-galactosyl ester linkage by beta-galactosidases. the enzyme from penicillium multicolor hydrolyzed the substrate as fast as p-nitrophenyl beta-galactoside (pnp-gal), a usual substrate with a beta-galactosidic linkage. the enzymes from escherichia coli and aspergillus oryzae hydrolyzed phb-gal with almost the same rates as pnp-gal. the enzymes from bacillus circulans, saccharomyces frag ...200010993159
molecular cloning of genomic dna for fructose-1,6-bisphosphatase. from aspergillus oryzae.we have cloned and sequenced an aspergillus oryzae genomic dna fragment that encodes a fructose-1,6-bisphosphatase gene (fbpa) with the aim of studying transcriptional regulation mechanisms involved in basic metabolism. expression of fbpa was repressed in the presence of glucose, but not in the presence of pyruvate or sodium acetate in the medium. the crea and facb element found in the fbpa 5'-flanking region may be important in fbpa regulation.200010993168
release characteristics of a short-chain fatty acid, n-butyric acid, from its beta-cyclodextrin ester conjugate in rat biological media.6(a)-o-(n-butanoyl)-beta-cyclodextrin was prepared and its hydrolysis behavior in aqueous solutions and in rat intestinal fluids was investigated. furthermore, the enzymatic hydrolyses of the n-butyric acid-beta-cyclodextrin conjugate using alpha-amylase and esterase were studied to gain insight into the release behavior of n-butyric acid from the conjugate. the hydrolysis of the conjugate proceeded according to a first-order kinetics in aqueous solution, and gave a v-shaped ph profile, indicati ...200011015693
mycelium-bound carboxylesterase from aspergillus oryzae: an efficient catalyst for acetylation in organic solvent.dry mycelium of a strain of aspergillus oryzae efficiently catalyzed the esterification between free acetic acid and primary alcohols (geraniol and ethanol) in organic solvent. the growth conditions to obtain high activity of mycelium-bound enzymes were firstly evaluated. a medium containing tween 80 as carbon source furnished mycelium with the highest activity in the hydrolysis of alpha-naphthyl esters (alpha-n-acetate, butyrate, caprylate). dry mycelium was employed to select suited conditions ...200011024527
synthesis and enzymatic evaluation of five-membered iminocyclitols and a pseudodisaccharide.described here are the synthesis of five-membered iminocyclitols with galacto-configuration and a pseudodisaccharide, and their inhibitory activities against beta-galactosyltransferase, beta-galactosidase and alpha-mannosidase.200011026538
aspergillus fumigatus alb1 encodes naphthopyrone synthase when expressed in aspergillus oryzae.conidial pigment biosynthesis is an important virulence factor in aspergillus fumigatus, a human fungal pathogen. involvement of dhn-melanin pathway in the biosynthesis of a. fumigatus conidial pigment implies that the alb1p polyketide synthase (pks) is a 1,3,6, 8-tetrahydroxynaphthalene (t4hn) synthase. the alb1p, however, shows higher sequence similarity to a naphthopyrone synthase than to a t4hn synthase. to clarify the function of alb1p, the alb1 gene was overexpressed in a heterologous host ...200011040426
determination of depolymerization kinetics of amylose, amylopectin, and soluble starch by aspergillus oryzae alpha-amylase using a fluorimetric 2-p-toluidinylnaphthalene-6-sulfonate/flow-injection analysis system.this study reports on the determination of the depolymerization kinetics of amylose, amylopectin, and soluble starch by aspergillus oryzae alpha-amylase using flow-injection analysis with fluorescence detection and 2-p-toluidinylnaphthalene-6-sulfonate as the fluorescent probe. the experimental data points, corresponding to the evolution of the concentration of "detectable" substrate with depolymerization time, were fit to a single exponential decay curve in the case of amylose and to a double e ...200011042551
purification and characterization of chitosanase and exo-beta-d-glucosaminidase from a koji mold, aspergillus oryzae iam2660.chitosan-degrading activity was detected in the culture fluid of aspergillus oryzae, a. sojae, and a. flavus among various fungal strains belonging to the genus aspergillus. one of the strong producers, a. oryzae iam2660 had a higher level of chitosanolytic activity when n-acetylglucosamine (glcnac) was used as a carbon source. two chitosanolytic enzymes, 40 kda and 135 kda in molecular masses, were purified from the culture fluid of a. oryzae iam2660. viscosimetric assay and an analysis of reac ...200011055393
temperature control in a continuously mixed bioreactor for solid-state fermentation.a continuously mixed, aseptic paddle mixer was used successfully for solid-state fermentation (ssf) with aspergillus oryzae on whole wheat kernels. continuous mixing improved temperature control and prevented inhomogeneities in the bed. respiration rates found in this system were comparable to those in small, isothermal, unmixed beds, which showed that continuous mixing did not cause serious damage to the fungus or the wheat kernels. continuous mixing improves heat transport to the bioreactor wa ...200111114659
model for on-line moisture-content control during solid-state this study we describe a model that estimates the extracellular (nonfungal) and overall water contents of wheat grains during solid-state fermentation (ssf) with aspergillus oryzae, using on-line measurements of oxygen, carbon dioxide, and water vapor in the gas phase. the model uses elemental balances to predict substrate dry matter losses from carbon dioxide measurements, and metabolic water production, water used in starch hydrolysis, and water incorporated in new biomass from oxygen measu ...200111114660
new syntheses of 1d- and 1l-1,2-anhydro-myo-inositol and assessment of their glycosidase inhibitory activities.the 1d and 1l enantiomers of 1,2-anhydro-myo-inositol (conduritol b epoxide) were synthesised from 1d-pinitol and 1l-quebrachitol, respectively, and their activities were compared in selected glycosidase inhibition assays. the 1d enantiomer was found to be the active isomer, functioning as an irreversible inhibitor of sweet almond beta-d-glucosidase. neither isomer was active against the alpha-d-glucosidase from bacillus stearothermophilus or the beta-d-galactosidase from aspergillus oryzae.200011117313
genetic analysis of growth inhibition of yeast cells caused by expression of aspergillus oryzae rnase t1.even though most fungal hydrolytic enzymes have been successfully secreted in s. cerevisiae cells by expression of corresponding cdna, overexpression of a. oryzae rnase t1 causes severe growth inhibition in yeast. we observed that yeast strains carrying rnase t1 cdna under control of the gal1 promoter with a single-copy vector were able to grow on galactose medium while those with a multi-copy vector were not. it was found that overexpression of three mutated versions of rnase t1 with low enzyma ...200011129588
cloning and expression in phospholipid containing cultures of the gene encoding the specific phosphatidylglycerol/phosphatidylinositol transfer protein from aspergillus oryzae: evidence that the pg/pi-tp is tandemly arranged with the putative 3-ketoacyl-coa thiolase gene.the phosphatidylglycerol/phosphatidylinositol transfer protein (pg/pi-tp) is a new and original phospholipid transfer protein (pltp) isolated from the deuteromycete, aspergillus oryzae. we have isolated a genomic clone of the a. oryzae pg/pi-tp using a probe derived from the corresponding cdna and sequenced the complete gene. the dna sequence analysis revealed that pg/pi-tp gene is composed of three exons encoding a 18,823 da protein of 175 amino acids as previously described and of two introns ...200111179668
cloning of the aspergillus oryzae 5-aminolevulinate synthase gene and its use as a selectable marker.the hema gene encoding 5-aminolevulinate synthase, the first enzyme in heme biosynthesis, was cloned from aspergillus oryzae and evaluated as a selectable marker for the transformation of filamentous fungi. deletion of the hema gene resulted in a lethal phenotype that could be rescued either by the supplementation of culture media with 5-aminolevulinic acid (ala) or by transformation with the wild-type hema gene, but not by growth on rich media, nor by the addition of exogenous heme. transformat ...200011191214
molecular cloning, overexpression, and purification of a major xylanase from aspergillus oryzae.the gene encoding xylanase g2 (xyng2) was isolated from a genomic library of aspergillus oryzae kbn616, used for making shoyu koji. the structural part of xyng2 was found to be 767 bp. the nucleotide sequence of cdna amplified by rt-pcr showed that the open reading frame of xyng2 was interrupted by a single intron which was 71 bp in size and encoded 232 amino acids. direct n-terminal amino acid sequencing showed that the precursor of xyng2 had a signal peptide of 44 amino acids. the predicted am ...200011210150
alpha-amylase production in high cell density submerged cultivation of aspergillus oryzae and a. nidulans.the effect of biomass concentration on the formation of aspergillus oryzae alpha-amylase during submerged cultivation with a. oryzae and recombinant a. nidulans strains has been investigated. it was found that the specific rate of alpha-amylase formation in chemostats decreased significantly with increasing biomass concentration in the range of approx. 2-12 g dry weight kg(-1). when using a recombinant a. nidulans strain in which the gene responsible for carbon catabolite repression of the a. or ...200111234963
the aspergillus nidulans multimeric ccaat binding complex ancf is negatively autoregulated via its hapb subunit gene.cis-acting ccaat elements are frequently found in eukaryotic promoter regions. many of them are bound by conserved multimeric complexes. in the fungus aspergillus nidulans the respective complex was designated ancf (a. nidulans ccaat binding factor). ancf is composed of at least three subunits designated hapb, hapc and hape. here, we show that the promoter regions of the hapb genes in both a. nidulans and aspergillus oryzae contain two inversely oriented, conserved ccaat boxes (box alpha and box ...200111243777
a cardinal model to describe the effect of water activity on the growth of moulds.a simple model was proposed to describe the effect of water activity (aw) on the radial growth rate of moulds. this model is deduced from the cardinal model family proposed by rosso in 1995, which is only defined from cardinal values of environmental factors (minimum, optimum and maximum values), the growth rate observed at the optimal value of the environmental factors, and n, a shape parameter. for aw, a simple form of cardinal model is proposed. this form is obtained for n = 2 and aw(max) = 1 ...200111246910
sequence analysis and overexpression of a pectin lyase gene (pel1) from aspergillus oryzae kbn616.a gene (pel1) encoding pectin lyase (pel1) was isolated from a shoyu koji mold, aspergillus oryzae kbn616, and characterized. the structural gene comprised 1,196 bp with a single intron. the orf encoded 381 amino acids with a signal peptide of 20 amino acids. the deduced amino acid sequence showed high similarity to those of aspergillus niger pectin lyases and glomerella cingulata pnla. the pel1 gene was successfully overexpressed under the promoter of the a. oryzae tef1 gene. the molecular mass ...200111272833
high-level secretory production of phospholipase a1 by saccharomyces cerevisiae and aspergillus oryzae.phospholipase a1 (pla1) is a hydrolytic enzyme that catalyzes the removal of the acyl group from position 1 of lecithin to form lysolecithin. the pla1 gene, which had been cloned from aspergillus oryzae, was expressed in saccharomyces cerevisiae and a. oryzae. through the modification of the medium composition and the feeding conditions of substrate, the production level of pla1 by s. cerevisiae was increased to a level fivefold higher than that indicated in a previous report. in the case of a. ...200111272851
the phylogenetics of mycotoxin and sclerotium production in aspergillus flavus and aspergillus oryzae.aspergillus flavus is a common filamentous fungus that produces aflatoxins and presents a major threat to agriculture and human health. previous phylogenetic studies of a. flavus have shown that it consists of two subgroups, called groups i and ii, and morphological studies indicated that it consists of two morphological groups based on sclerotium size, called "s" and "l." the industrially important non-aflatoxin-producing fungus a. oryzae is nested within group i. three different gene regions, ...200011273679
synergistic action of an x-prolyl dipeptidyl aminopeptidase and a non-specific aminopeptidase in protein hydrolysis.non-specific monoaminopeptidase (ap; e.c. 3.4.11) and x-prolyl dipeptidyl aminopeptidase (x-pdap; e.c., both from aspergillus oryzae, demonstrate strong synergism in hydrolyzing proline-containing peptides. incubation of ap alone with the peptide ala-pro-gly-asp-arg-ile-tyr-val-his-pro-phe does not generate free amino acids. however, when ap and x-pdap are added in combination, complete and immediate hydrolysis of all peptide bonds, other than x-pro bonds, is observed. in the enzymatic ...200111308367
isoenzyme multiplicity and characterization of recombinant manganese peroxidases from ceriporiopsis subvermispora and phanerochaete chrysosporium.we expressed cdnas coding for manganese peroxidases (mnps) from the basidiomycetes ceriporiopsis subvermispora (mnp1) and phanerochaete chrysosporium (h4) under control of the alpha-amylase promoter from aspergillus oryzae in aspergillus nidulans. the recombinant proteins (rmnp1 and rh4) were expressed at similar levels and had molecular masses, both before and after deglycosylation, that were the same as those described for the mnps isolated from the corresponding parental strains. isoelectric ...200111319083
enzymatic glycosylation using 6-o-acylated sugar donors and acceptors: beta-n-acetylhexosaminidase-catalysed synthesis of 6-o,n,n'-triacetylchitobiose and 6'-o,n,n'-triacetylchitobiose.p-nitrophenyl 6-o-acetyl-2-acetamido-2-deoxy-beta-d-glucopyranoside (5a) was used as the glycosyl donor in a beta-n-acetylhexosaminidase-catalysed (from penicillium brasilianum) glycosylation of glcnac yielding 6'-o,n,n'-triacetylchitobiose (6), while 6-o-acetyl-2-acetamido-2-deoxy-beta-d-glucopyranose (3a) served as a selectively protected acceptor in a transglycosylation reaction catalysed by the same enzyme to yield 6-o,n,n'-triacetylchitobiose (4).200111322728
stimulation of myelopoiesis in listeria monocytogenes-infected mice by an aggregated polymer isolated from aspergillus this work, we investigated the effects of the proteic aggregated polymer of magnesium ammonium phospholinoleate-palmitoleate anhydride (mapa) isolated from aspergillus oryzae on the growth and differentiation of bone marrow granulocyte-macrophage progenitor cells (cfu-gm) in listeriamonocytogenes-infected mice. a significant reduction in the cfu-gm number was observed in the initial phase of infection with a sublethal dose of listeria. treatment of mice with 0.5, 2.0 and 5.0 mg/kg mapa for 7 ...200111339624
cloning and characterization of a chitosanase gene from the koji mold aspergillus oryzae strain iam 2660.a genomic copy of the gene coding for chitosanase (csna) was isolated from aspergillus oryzae iam 2660. a. oryzae csna contains an open reading frame that encodes a polypeptide of 245 amino acids with a calculated molecular mass of 26,500 da. the deduced amino acid sequence of a. oryzae csna indicates extensive similarities to those of other fungal chitosanases.200111388486
production and properties of beta-mannanase by free and immobilized cells of aspergillus oryzae nrrl fungi were tested for production of mannanases. the highest mannanase activities were produced by aspergillus oryzae nrrl 3488 after 7 days in static cultures. mannanases were induced by gum locust bean (1.0%). the highest mannanase activity was produced when a mixture of peptone, urea and ammonium sulphate was used as nitrogen source. zn2+ or co2+ favoured enzyme production. the immobilized cells on ca-alginate and agar were able to produce beta-mannanase for four runs with a slight decre ...200111393772
single-stranded regions in dna isolated from different developmental stages of the sea urchin.long-term labeled sea urchin embryo (strongylocentrotus purpuratus) dnas were examined for size of recovered pieces, single-strandedness, and length of continuous double-stranded regions. sizing on neutral sucrose gradients indicates that morula stage dna sediments predominantly at 31 s, blastula stage dna at 27 s, and gastrula stage dna as a broad range of sizes of greater than 29 s. treatment of [3h]thymidine-labeled dna with aspergillus oryzae s1 nuclease removes 19% of the 3h from morula sta ...197411400427
isolation of a short, cytosine-rich repeating unit from the dna of escherichia coli.hybridization of heterologous nucleic acids has provided the means for isolating a repeating sequence which is located next to template regions of dna. separated single strands of 32p-labelled dna from escherichia coli were to a limited extent able to anneal with dna of micrococcus lysodeikticus immobilized on nitrocellulose membrane filters. the resulting hybrid was resistant to enzymes specific for unpaired strands, nuclease s1 (aspergillus oryzae) and exonuclease i (e. coli). the e. coli dna ...197411400431
aohapb, aohapc and aohape, subunits of the aspergillus oryzae ccaat-binding complex, are functionally interchangeable with the corresponding subunits in aspergillus nidulans.two genes, aohapb and aohape, encoding subunits of the aspergillus oryzae ccaat-binding complex were cloned and sequenced. the polypeptides encoded by aohapb and aohape were expressed in escherichia coli and used to reconstitute a dna-binding complex with recombinant aohapc. the dna-binding activity was observed only in the presence of all three subunits, indicating that aohapb, aohape and aohapc are essential for ccaat-binding. furthermore, introduction of the aohapb, aohapc and aohape genes in ...200111409179
enzymatic synthesis of two ginsenoside re-beta-xylosides.two new beta-xylosyl derivatives of ginsenoside re, 20(s)-protopanaxatriol 6-o-alpha-l-rhamnopyranosyl-(1 --> 2)-[beta-d-xylopyranosyl-(1 --> 4)]-beta-d-glucopyranosyl-20-o-beta-d-glucopyranoside and 20(s)-protopanaxatriol 6-o-alpha-l-rhamnopyranosyl-(1 --> 2)-[beta-d-xylopyranosyl-(1 --> 6)]-beta-d-glucopyranosyl-20-o-beta-d-glucopyranoside, were respectively synthesized from p-nitrophenyl beta-d-xylopyranoside and phenyl beta-d-xylopyranoside as donors and ginsenoside re as the acceptor in 25% ...200111440145
[mystery of fermented foods--there are friendly molds too!]. 200111441805
filamentous fungus aspergillus oryzae has two types of alpha-1,2-mannosidases, one of which is a microsomal enzyme that removes a single mannose residue from man9glcnac2.alpha-mannosidase activities towards high-mannose oligosaccharides were examined with a detergent-solubilized microsomal preparation from a filamentous fungus, aspergillus oryzae. in the enzymatic reaction, the pyridylaminated substrate man(9)glcnac(2)-pa was trimmed to man(8)glcnac(2)-pa which lacked one alpha-1,2-mannose residue at the nonreducing terminus of the middle branch (man8b isomer), and this mannooligosaccharide remained predominant through the overall reaction. trimming was optimal ...200011443275
functional analysis of tama, a coactivator of nitrogen-regulated gene expression in aspergillus nidulans.the tam a gene of aspergillus nidulans encodes a 739-amino acid protein with similarity to uga35p/dal81p/durlp of saccharomyces cerevisiae. it has been proposed that tama functions as a co-activator of area, the major nitrogen regulatory protein in a. nidulans. because area functions as a transcriptional activator under nitrogen-limiting conditions, we investigated whether tama was also present in the nucleus. we found that a gfp-tama fusion protein was predominantly localised to the nucleus in ...200111459183
screening and characterization of koji molds producing saline-tolerant protease.three mold strains isolated from soil in the taipei area of taiwan were compared with a commercial strain of aspergillus oryzae for their proteolytic activities in an 18% nacl aqueous solution system. among these strains, the one subsequently identified and designated as aspergillus sp. fc-10 produced protease with superior saline tolerance. in aflatoxin tests, this strain did not generate detectable aflatoxin after growing on steamed grain polished rice substrate for 24 days. two types of extra ...200111464272
bi-fluorescence-labeled maltoheptaoside: convenient substrate for continual assay of alpha-amylase.a new maltoheptaose derivative was prepared as a useful substrate for continual assay of alpha-amylase. the maltoheptaoside has thionaphtyl group as a fluorescent energy donor at the reducing end and dansyl group as an acceptor group at the non-reducing end. excitation of the thionaphthyl group at 290 nm results in emission at 523 nm from the dansyl group, while the emission from the thionaphthyl group is quenched by the dansyl group. this fluorescence energy transfer is reduced by the hydrolyti ...200111506161
isoflavone transformation during soybean koji preparation and subsequent miso fermentation supplemented with ethanol and nacl.soybeans were soaked with water for 4 h, steam-cooked, inoculated with the conidia of aspergillus oryzae, and incubated for 3 days for koji preparation. the koji was then mixed with water-soaked and steam-cooked soybeans (1:2, w/w), ground into paste, and supplemented with 15% ethanol and 12.5% nacl or 3% ethanol and 6% nacl for miso fermentation at 30 degrees c. daidzin, genistin, daidzein, and genistein contents were extracted from the lyophilized and pulverized soybean powder or from the miso ...200111513643
visualization of nuclei in aspergillus oryzae with egfp and analysis of the number of nuclei in each conidium by facs.aspergillus oryzae has been reported to form conidia with multinuclei. in order to analyze nuclei in living cells, we developed an expression system of the a. nidutans histone h2b protein tagged by egfp (h2b::egfp). in both a. oryzae niad300 and a. nidulans fgsc89 transformants expressing h2b::egfp, fluorescence was detected in nuclear regions of hyphae and conidia. while a conidium contained only one fluorescent spot in the a. nidulans transformant, approximately 66% of conidia had two, 24% had ...200111515532
a new class of glutaminase from aspergillus oryzae.the koji mold aspergillus oryzae is able to produce glutaminase which converts glutamine to glutamic acid, one of the most important flavor components in soy sauce. we present here the isolation and the complete nucleotide sequence of the glutaminase- encoding gene from a. oryzae u212, an industrial strain used in thailand. n-terminal and internal amino acid sequences were determined from purified glutaminase. a 700-bp fragment was amplified by pcr using oligonucleotide primers designed from par ...200111545278
increased phospholipid transfer protein activity in aspergillus oryzae grown on various industrial phospholipid sources.the effect of industrial carbon sources on phospholipid transfer protein production was investigated. phospholipid fractions of different composition were prepared from various plant oils (i.e., soybean, rapeseed, and sunflower) according to the lucas meyer extraction and purification process. the effect of these fractions on phospholipid transfer protein activity of cell extracts from aspergillus oryzae grown on medium containing these phospholipids as sole carbon source was studied. it was sho ...200111547891
on the involvement of electron transfer reactions in the fluorescence decay kinetics heterogeneity of proteins.time-resolved fluorescence study of single tryptophan-containing proteins, nuclease, ribonuclease t1, protein g, glucagon, and mastoparan, has been carried out. three different methods were used for the analysis of fluorescence decays: the iterative reconvolution method, as reviewed and developed in our laboratory, the maximum entropy method, and the recent method that we called "energy transfer" method. all the proteins show heterogeneous fluorescence kinetics (multiexponential decay). the orig ...200111567101
expression, gene cloning, and characterization of five novel phytases from four basidiomycete fungi: peniophora lycii, agrocybe pediades, a ceriporia sp., and trametes pubescens.phytases catalyze the hydrolysis of phosphomonoester bonds of phytate (myo-inositol hexakisphosphate), thereby creating lower forms of myo-inositol phosphates and inorganic phosphate. in this study, cdna expression libraries were constructed from four basidiomycete fungi (peniophora lycii, agrocybe pediades, a ceriporia sp., and trametes pubescens) and screened for phytase activity in yeast. one full-length phytase-encoding cdna was isolated from each library, except for the ceriporia sp. librar ...200111571175
fluorescence polarization: analysis of carbohydrate-protein interaction.fluorescence polarization has been widely used for the studies on the molecular motion in solution and has been applied to immunoassays for proteins, therapeutic drug monitoring in clinical pharmacy, and assays for environmentally toxic compounds. because fluorescence polarization is most readily applicable to the kinetic analysis of the binding reaction between a substance having small molecular mass and a receptor molecule, this method is easily applied to the analysis of carbohydrate-lectin b ...200111673876
identification of catalytic and substrate-binding site residues in bacillus cereus atcc7064 oligo-1,6-glucosidase.three active site residues (asp199, glu255, asp329) and two substrate-binding site residues (his103, his328) of oligo-1,6-glucosidase (ec from bacillus cereus atcc7064 were identified by site-directed mutagenesis. these residues were deduced from the x-ray crystallographic analysis and the comparison of the primary structure of the oligo-1,6-glucosidase with those of saccharomyces carlsbergensis alpha-glucosidase, aspergillus oryzae alpha-amylase and pig pancreatic alpha-amylase which ...200111676021
a quick solution: ab initio structure determination of a 19 kda metalloproteinase using acorn.a data set from the metalloproteinase deuterolysin was collected at atomic resolution (1.0 a) with synchrotron radiation. the high resolution allowed the structure to be solved with the new direct-methods program acorn using the coordinates of the zn atom as a starting point. the phases obtained from acorn were of sufficient quality to allow automated building to be carried out in arp/warp. minimal manual rebuilding of the model was required and the structure determination was completed using th ...200111679721
cloning and sequence analysis of cnaa gene encoding the catalytic subunit of calcineurin from aspergillus oryzae.calcineurin has been implicated in ion-homeostasis, stress adaptation in yeast and for hyphal growth in filamentous fungi. genomic dna and cdna encoding the catalytic subunit of calcineurin (cnaa) were isolated from aspergillus oryzae. the cnaa open reading frame extended to 1727 bp and encoded a putative protein of 514 amino acids. comparative analysis of the nucleotide sequence of cnaa genomic dna and cdna confirmed the presence of three introns and a highly conserved calmodulin binding domain ...200111682197
physiological characterisation of recombinant aspergillus nidulans strains with different crea genotypes expressing a. oryzae alpha-amylase.the physiology of three strains of aspergillus nidulans was examined--a crea deletion strain, a wild type crea genotype and a strain containing extra copies of the crea gene, all producing aspergillus oryzae alpha-amylase. the strains were cultured in batch and continuous cultivations and the biomass formation and alpha-amylase production was characterised. overexpression of the crea gene resulted in a lower maximum specific growth rate and a slightly higher repression of the alpha-amylase produ ...200211689252
establishment of a hyper-protein production system in submerged aspergillus oryzae culture under tyrosinase-encoding gene (melo) promoter control.uv-mediated mutagenesis generated a high glucoamylase-producing mutant of aspergillus oryzae exhibiting strong melanization in solid-state culture. expression of the glucoamylase-encoding gene (glab), which is specifically expressed in solid-state culture, and the tyrosinase-encoding gene (melo), was analyzed using an e. coli beta-glucuronidase (gus) reporter assay to investigate this phenomenon. although no common regulation was found for melo and glab expression, the former was greatly enhance ...200111693910
electrophoretic mobility shift scanning using an automated infrared dna sequencer.electrophoretic mobility shift assay (emsa) is widely used in the study of sequence-specific dna-binding proteins, including transcription factors and mismatch binding proteins. we have established a non-radioisotope-based protocol for emsa that features an automated dna sequencer with an infrared fluorescent dye (irdye) detection unit. our modification of the elec- trophoresis unit, which includes cooling the gel plates with a reduced well-to-read length, has made it possible to detect shifted ...200111730013
production of galacto-oligosaccharides from lactose by aspergillus oryzae beta-galactosidase immobilized on cotton cloth.the production of galacto-oligosaccharides (gos) from lactose by a. oryzae beta-galactosidase immobilized on cotton cloth was studied. the total amounts and types of gos produced were mainly affected by the initial lactose concentration in the reaction media. in general, more and larger gos can be produced with higher initial lactose concentrations. a maximum gos production of 27% (w/w) of initial lactose was achieved at 50% lactose conversion with 500 g/l of initial lactose concentration. tri-s ...200211745169
reactivity and stability of mycelium-bound carboxylesterase from aspergillus oryzae.the reactivity and thermostability of a novel mycelium-bound carboxylesterase from lyophilized cells of aspergillus oryzae are explored in organic solvent. ethanol acetylation was selected as reference esterification reaction. high carboxylesterase activity cells were used as biocatalyst in batch esterification tests at 12.5 < s(o) < 125 mmol l(-1), 5.0 < x(o) < 30 g l(-1), 0.49 < log p < 4.5 and 30 < t < 80 degrees c, as well as in residual activity tests after incubation at 40 < t < 90 degrees ...200211753931
influence of carbon source on alpha-amylase production by aspergillus oryzae.the influence of the carbon source on alpha-amylase production by aspergillus oryzae was quantified in carbon-limited chemostat cultures. the following carbon sources were investigated: maltose, maltodextrin (different chain lengths), glucose, fructose, galactose, sucrose, glycerol, mannitol and acetate. a. oryzae did not grow on galactose as the sole carbon source, but galactose was co-metabolized together with glucose. relative to that on low glucose concentration (below 10 mg/l), productivity ...200111759683
a novel thermostable branching enzyme from an extremely thermophilic bacterial species, rhodothermus obamensis.a branching enzyme (ec gene was isolated from an extremely thermophilic bacterium, rhodothermus obamensis. the predicted protein encodes a polypeptide of 621 amino acids with a predicted molecular mass of 72 kda. the deduced amino acid sequence shares 42-50% similarity to known bacterial branching enzyme sequences. similar to the bacillus branching enzymes, the predicted protein has a shorter n-terminal amino acid extension than that of the escherichia coli branching enzyme. the deduce ...200111778874
a new flavone glycoside, 5-hydroxy 7,3',4',5'-tetra-methoxyflavone 5-o-beta-d-xylopyranosyl-(1-->2)-alpha-l-rhamnopyranoside from bauhinia variegata linn.a new flavone glycoside m.f. c(30)h(36)o(15) m.p. 252-253 degrees c, [m]+ 636 (eims) was isolated from the acetone soluble fraction of the concentrated 95% ethanolic extract of the seeds of bauhinia variegata (linn). it was identified as 5-hydroxy7,3',4',5'-tetra-methoxyflavone 5-o-beta-d-xylopyranosyl-(1-->2)-alpha-l-rhamnopyranoside (1) by various colour reactions, chemical degradations and spectral techniques.200111783588
observation of egfp-visualized nuclei and distribution of vacuoles in aspergillus oryzae arpa null mutant.the arpa gene encoding arp1 (actin-related protein) was previously cloned and characterized from aspergillus oryzae. phenotypes of the arpa null mutant indicate its requirement for normal nuclear distribution and morphology of conidiophores. in this study, we further characterized the function of the arpa gene in distribution of organelles. for further analysis of nuclear migration in living cells, an expression system consisting of a fusion protein of aspergillus nidulans histone h2b and egfp ( ...200211786257
validation of a model for process development and scale-up of packed-bed solid-state bioreactors.we have validated our previously described model for scale-up of packed-bed solid-state fermenters (weber et al., 1999) with experiments in an adiabatic 15-dm(3) packed-bed reactor, using the fungi coniothyrium minitans and aspergillus oryzae. effects of temperature on respiration, growth, and sporulation of the biocontrol fungus c. minitans on hemp impregnated with a liquid medium were determined in independent experiments, and the first two effects were translated into a kinetic model, which w ...200211787011
deletion analysis of the enolase gene (enoa) promoter from the filamentous fungus aspegillus oryzae.the enolase gene (enoa) is one of the most strongly expressed genes in aspergillus oryzae. to elucidate the transcription regulatory element for this strong expression and the process of glucose induction, the transcription activity of a series of truncated enoa promoters was measured by using the escherichia coli uida gene as a reporter. deletion of a 104-bp region located -224 nt to -121 nt upstream of the translation initiation site caused both a drastic decrease in the beta-glucuronidase (gu ...200111795846
plasminogen-binding activity of enolase in the opportunistic pathogen pneumocystis carinii.the glycolytic enzyme enolase is one of the most abundant proteins expressed in fungi and has been shown to be an immunodominant cell-wall-associated antigen of the pathogenic fungus, candida albicans. enolase has also been found on the surface of some mammalian cells where it functions as a plasminogen-binding motif and facilitator of plasminogen activation to plasmin. to investigate the immunogenicity of enolase in the opportunistic pathogen, pneumocystis carinii, the genomic and complementary ...200111798055
estimation of hyphal tensile strength in production-scale aspergillus oryzae fungal fermentations.fragmentation of filamentous fungal hyphae depends on two phenomena: hydrodynamic stresses, which lead to hyphal breakage, and hyphal tensile strength, which resists breakage. the goal of this study was to use turbulent hydrodynamic theory to develop a correlation that allows experimental data of morphology and hydrodynamics to be used to estimate relative (pseudo) tensile strength (sigma(pseudo)) of filamentous fungi. fed-batch fermentations were conducted with a recombinant strain of aspergill ...200211807755
behaviors of d- and l-lactic acids during the brewing process of sake (japanese rice wine).the amounts of d- and l-lactic acids during the brewing process of sake were determined by capillary electrophoresis using 2-hydroxypropyl-beta-cyclodextrin as a chiral selector. because l-lactic acid, which prevents the growth of nonuseful microorganisms, is a raw material of sake, the ratio of l-lactic acid to total lactic acid is almost 1.0 at the initial stage of sake brewing. during brewing, the ratio decreased gradually and finally reached 0.39. yeast (saccharomyces cerevisiae) for sake br ...200211829643
intake, digestion, and digestive characteristics of neotyphodium coenophialum-infected and uninfected fescue by heifers offered hay diets supplemented with aspergillus oryzae fermentation extract or laidlomycin propionate.tarentaise heifers fitted with a rumen cannula (539 +/- 7.5 and 487 +/- 15.7 kg avg initial bw in exp. 1 and 2, respectively) were used in two latin square metabolism experiments having 2 x 2 factorial treatment arrangements to determine the effects of supplementation with aspergillus oryzae fermentation extract (ao) or laidlomycin propionate (lp) on intake, digestion, and digestive characteristics of neotyphodium coenophialum-infected (if) or uninfected (ff) tall fescue (festuca arundinacea) ha ...200211831521
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