TitleAbstractYear(sorted ascending)
purification and characterization of the overexpressed aspergillus oryzae xylanase, xynf1.the aspergillus oryzae xynf1 gene coding for a xylanase, xynf1, was successfully overexpressed under the strong a. oryzae tef1 gene promoter. the high-xynf1-producing transformant secreted about 180 mg/l of xynf1 in the glucose-containing medium. the overexpressed xynf1 was purified by only one chromatographic step. the purified xynf1 had a molecular mass of 35.0 kda, a ph optimum of 5.0, and a temperature optimum of 60 degrees c.199910586505
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
identification of enzymes and quantification of metabolic fluxes in the wild type and in a recombinant aspergillus oryzae straintwo alpha-amylase-producing strains of aspergillus oryzae, a wild-type strain and a recombinant containing additional copies of the alpha-amylase gene, were characterized with respect to enzyme activities, localization of enzymes to the mitochondria or cytosol, macromolecular composition, and metabolic fluxes through the central metabolism during glucose-limited chemostat cultivations. citrate synthase and isocitrate dehydrogenase (nad) activities were found only in the mitochondria, glucose-6-p ...19999872753
sequence analysis, overexpression, and antisense inhibition of a beta-xylosidase gene, xyla, from aspergillus oryzae kbn616.beta-xylosidase secreted by the shoyu koji mold, aspergillus oryzae, is the key enzyme responsible for browning of soy sauce. to investigate the role of beta-xylosidase in the brown color formation, a major beta-xylosidase, xyla, and its encoding gene were characterized. beta-xylosidase xyla was purified to homogeneity from culture filtrates of a. oryzae kbn616. the optimum ph and temperature of the enzyme were found to be 4.0 and 60 degrees c, respectively, and the molecular mass was estimated ...19999872754
homologs of aflatoxin biosynthesis genes and sequence of aflr in aspergillus oryzae and aspergillus sojae.the presence, but not expression, of homologs of three structural genes and a regulatory gene necessary for aflatoxin biosynthesis in aspergillus parasiticus and a. flavus was shown for a. oryzae and a. sojae. homologs of the regulatory gene aflr were cloned and sequenced from a. oryzae and a. sojae.19999872797
a xyloglucan-specific endo-beta-1,4-glucanase from aspergillus aculeatus: expression cloning in yeast, purification and characterization of the recombinant enzyme.a full-length c-dna encoding a xyloglucan-specific endo -beta-1, 4-glucanase (xeg) has been isolated from the filamentous fungus aspergillus aculeatus by expression cloning in yeast. the colonies expressing functional xeg were identified on agar plates containing azurine-dyed cross-linked xyloglucan. the cdna encoding xeg was isolated, sequenced, cloned into an aspergillus expression vector, and transformed into aspergillus oryzae for heterologous expression. the recombinant enzyme was purified ...19999884411
comparison of gene structures and enzymatic properties between two endoglucanases from humicola grisea.we have cloned two endoglucanase genes (egl3 and egl4) from a thermophilic fungus, humicola grisea. the coding region of the egl3 gene was interrupted by an intron of 56-bp, and the deduced amino acid sequence of the egl3 gene was 305 amino acids in length and showed 98.4% identity with humicola insolens egv. the coding region of the egl4 gene was also interrupted by an intron of 173-bp, which contains 34 ttc repeated sequence units, and the deduced amino acid sequence of the egl4 gene was 227 a ...19999990729
characterization and expression of the cdna encoding a new kind of phospholipid transfer protein, the phosphatidylglycerol/phosphatidylinositol transfer protein from aspergillus oryzae: evidence of a putative membrane targeted phospholipid transfer protein in fungi.the full-length cdna of a phospholipid transfer protein (pltp) was isolated from aspergillus oryzae by a race-pcr procedure using degenerated primer pool selected from the n-terminal sequence of the purified phosphatidylinositol/phosphatidylglycerol transfer protein (pg/pi-tp). the cdna encodes a 173 amino acid protein of 18823 da. the deduced amino acid sequence from position 38 to 67 is 100% identical to the n-terminal sequence (first 30 amino acids) of the purified pg/pi-tp. this amino acid s ...199910023082
three-dimensional structure of ribonuclease t1 complexed with an isosteric phosphonate substrate analogue of gpu: alternate substrate binding modes and catalysis.the x-ray crystal structure of a complex between ribonuclease t1 and guanylyl(3'-6')-6'-deoxyhomouridine (gpcu) has been determined at 2. 0 a resolution. this ligand is an isosteric analogue of the minimal rna substrate, guanylyl(3'-5')uridine (gpu), where a methylene is substituted for the uridine 5'-oxygen atom. two protein molecules are part of the asymmetric unit and both have a gpcu bound at the active site in the same manner. the protein-protein interface reveals an extended aromatic stack ...199910029539
cloning and expression of a cdna encoding the laccase from schizophyllum commune.we cloned and analyzed the nucleotide sequence of a cdna that encodes polyphenol oxidase (laccase) from the white-rot basidiomycete schizophyllum commune. the nucleotide sequence of the full-length cdna contained a 1554-base open reading frame that encoded a polypeptide of 518 amino acid residues, including a putative signal peptide of 16 residues. it contained four highly similar regions that are conserved in the deduced amino acid sequences of other laccases, including the region thought to be ...199910052122
pectin methylesterase gene (pmea) from aspergillus oryzae kbn616: its sequence analysis and overexpression, and characterization of the gene product.a gene (pmea) encoding pectin methylesterase was isolated from a shoyu koji mold, aspergillus oryzae kbn616, and characterized. the structural gene comprised 1,370 bp with six introns. the pmea protein consisted of 331 amino acids with a putative signal peptide of 17 amino acids. the deduced amino acid sequence was very similar to those of aspergillus niger pmea and aspergillus aculeatus pme1. the pmea gene was efficiently expressed under control of the a. oryzae tef1 gene promoter for purificat ...199910052131
insertion analysis of putative functional elements in the promoter region of the aspergillus oryzae taka-amylase a gene (amyb) using a heterologous aspergillus nidulans amds-lacz fusion gene system.expression of the taka-amylase a gene (amyb) of aspergillus oryzae is induced by starch or maltose. the a. oryzae amyb gene promoter contains three highly conserved sequences, designated regions i, ii, and iii, compared with promoter regions of the a. oryzae glaa encoding glucoamylase and the agda encoding alpha-glucosidase. to identify the function of these sequences within the amyb promoter, various fragments containing conserved sequences in the amyb promoter were introduced into the upstream ...199910052139
myrothecium verrucaria bilirubin oxidase and its mutants for potential copper ligands.bilirubin oxidase (ec: purified from a culture medium of myrothecium verrucaria mt-1 (authentic enzyme) catalyzes the oxidation of bilirubin to biliverdin in vitro and recombinant enzyme (wild type) was obtained by using an overexpression system of the bilirubin oxidase gene with aspergillus oryzae harboring an expression vector. the absorption and esr spectra showed that both bilirubin oxidases are multicopper oxidases containing type 1, type 2, and type 3 coppers similar to laccase, as ...199910074356
delta 9-fatty acid desaturase from arachidonic acid-producing fungus. unique gene sequence and its heterologous expression in a fungus, aspergillus.based on the sequence information for delta 9-desaturase genes (from rat, mouse and yeast), which are involved in the desaturation of palmitic acid and stearic acid to palmitoleic acid and oleic acid, respectively, the corresponding cdna and genomic gene were cloned from the fungal strain, mortierella alpina 1s-4, which industrially produces arachidonic acid. there was a cytochrome b5-like domain linked to the carboxyl terminus of this mortierella desaturase, as also seen in the yeast delta 9-de ...199910091601
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
role of substrate concentration in mitosis and hyphal extension of aspergillus.the filamentous fungi aspergillus oryzae and a. niger grow by apical extension of multinucleate hyphae that are subdivided into compartments by cross-walls called septa. submerged cultivation, image analysis, and fluorescence microscopy were used to study the role of the carbon source on mitosis and hyphal extension in these fungi. in the two species of aspergillus, the length of the apical compartment, the number of nuclei in the apical compartment, and the hyphal diameter were regulated in res ...200010620754
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
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
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
molecular breeding of yeast with higher metal-adsorption capacity by expression of histidine-repeat insertion in the protein anchored to the cell wall.a fusion protein of hexa-histidine repeat (his) and glycosylphosphatidylinositol (gpi)-anchor region of saccharomyces cerevisiae cwp1 with aspergillus oryzae taka-amylase a (taa) was expressed on the yeast cell surface. the expressed fusion protein (taa-his-cwp1) was localized on the cell wall and demonstrated amylolytic activity. in comparison with the taa-cwp1 expressing strain, these cells exhibited 1.6- to 2.8-fold higher adsorbing capacity for cu(2+), ni(2+), and zn(2+).200012483584
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
molecular cloning of the isoamyl alcohol oxidase-encoding gene (mrea) from aspergillus oryzae.isoamyl alcohol oxidase (iaaod) is a novel enzyme that catalyzes the formation of isovaleraldehyde, which is the main component of mureka that gives sake an off-flavor (yamashita et al. biosci. biotechnol. biochem., 63, 1216-1222, 1999). we cloned the genomic dna sequence encoding iaaod from a koji mold, aspergillus oryzae, using a pcr-amplified dna fragment corresponding to the partial amino acid sequences of the purified protein as a probe. the cloned gene comprises 1903 bp of an open reading ...200016232791
production and product quality assessment of human hepatitis b virus pre-s2 antigen in submerged and solid-state cultures of aspergillus oryzae.pre-s2 is a diagnostically important antigen of human hepatitis b virus (hbv). in order to produce pre-s2 antigen in aspergillus oryzae, the gene [pre-s2]3, which encodes a tandemly triplicated repeat of pre-s2 polypeptides was fused with the partial glaa gene encoding glucoamylase lacking the starch-binding domain. in submerged culture, a. oryzae transformants carrying glaa-[pre-s2]3 secreted a heterogeneously glycosylated form of the fusion protein that was partially degraded. contrarily, util ...200016232829
accumulation of a recombinant aspergillus oryzae lipase artificially localized on the bacillus subtilis cell surface.cutl cdna encoding an extracellular lipase, l1, from aspergillus oryzae was fused to the cell wall-binding domain (cwb) region of a plasmid, phcb3r. sds-polyacrylamide gel electrophoresis (page) and zymography of proteins extracted from the cell surface of bacillus subtilis 168 harboring a fused lipase plasmid (phcb3rcl) revealed that the fused gene product, cwb-cutl, was localized in the b. subtilis cell wall and retained lipase activity. b. subtilis wasd (wpra sigd), recently used for the accu ...200016232883
rapid detection of homologously integrated dna fragments and accurate quantitation of their copy number in transgenic aspergillus oryzae by pcr.a simple and rapid method for the analysis of artificially introduced dna fragments has been developed using competitive pcr and long and accurate pcr. the locus and the copy number of the dna fragments in each aspergillus oryzae transformant could be detected more rapidly and accurately by this method than by the conventional southern hybridization method.200016232915
improvement of production of kojic acid by a mutant strain aspergillus oryzae, mk107-39.a strain designated mk107-39, producing kojic acid with a high yield, was obtained by a new screening method using a 96-well microtiter plate after ntg treatment of aspergillus oryze atcc 22788. the amount of kojic acid produced by strain mk107-39 in a shaking flask was 28 g/l from 100 g/l of glucose, which was 7.7-times higher than that produced by parent strain. the kojic acid yields per cell and the amount of glucose consumed were 9.8 and 6.0-times higher than those of the parent strain. base ...200116232988
a second pectin lyase gene (pel2) from aspergillus oryzae kbn616: its sequence analysis and overexpression, and characterization of the gene products.a second pectin lyase gene, designated pel2, was isolated from a shoyu koji mold aspergillus oryzae kbn616 and characterized. the structural gene comprised 1306 bp with three introns. the orf encoded 375 amino acids with a signal peptide of 19 amino acids. the deduced amino acid sequence showed high similarity to those of a. oryzae pel1, aspergillus niger pectin lyases and glomerella cingulata pn1a. the pel2 gene was overexpressed under the control of the promoter of the a. oryzae tef1 gene for ...200116233008
overexpression and purification of aspergillus aculeatus beta-mannosidase and analysis of the integrated gene in aspergillus expression plasmid for the manb gene encoding aspergillus aculeatus beta-d-mannosidase (manb) was constructed by using an expression vector carrying an improved promoter. after transformation of a. oryzae by the plasmid, several transformants formed colonies emitting fluorescence on a plate containing 4-methylumbelliferyl beta-d-mannopyranoside (mu-man) under uv-irradiation. the transformant that displayed the strongest fluorescence, named a. oryzae bmn1, produced about 270 mg manb/l in liqui ...200116233072
kojic acid production in an airlift bioreactor using partially hydrolyzed raw corn a culture of aspergillus oryzae mk-107-39 in a 3-l airlift bioreactor, kojic acid was not produced when glucose/wheat germ medium (gm1) was used. however, when a jar fermentor was used, the kojic acid yield was high. a suitable medium for culture in an airlift bioreactor consisting of partially hydrolyzed corn starch and a small amount of corn steep liquor (csl) (sm1) was selected. in the cultivation in the airlift bioreactor using sm1, nearly 40 g/l of kojic acid was produced, which was the ...200116233111
efficient production of a heterologous peroxidase, dyp from geotrichum candidum dec 1, on solid-state culture of aspergillus oryzae rd005.the productivity of a peroxidase (dyp) originating from geotrichum candidum dec 1 was enhanced in the solid-state culture using aspergillus oryzae rd005. when the humidity, water content, and temperature were adjusted to 60%, 50% and 27 degrees c, respectively, the productivity of dyp reached 5.3 g per kilogram wheat bran, which was used as the solid medium. the yield of 5.3 g per kg wheat bran corresponded to the yield of a 56 kg submerged culture. the productivity per gram carbon of the medium ...200116233153
a new xylanase from penicillium griseofulvum.a new xylanase (pgxyna) from penicillium griseofulvum a160 has been isolated and characterised using a screening method based on the ability to digest a complex substrate. the enzyme belongs to the hydrolase family 11 or g and shows an optimum ph of 5.0 and an optimum temperature of 50 degrees c. the xylanase breaks down the xylan to very small oligosaccharides. the corresponding gene (pgxyna) was cloned and expressed in aspergillus oryzae. a second xylanase gene with 66% identity to pgxyna has ...200115954597
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
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
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
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
a new biologically active triterpenoid saponin from the leaves of lepidagathis hyalina nees.a new biologically active triterpenoid saponin m.f. c42h68o13, m.p. 315 degrees-317 degrees c was isolated from the ethyl acetate soluble fraction of the methanolic extract of the leaves of lepidagathis hyalina. its structure was characterized as 3-beta-o-[alpha-l-rhamnopyranosyl(1-->4)o-beta-d-glucopyranosyl]16-alpha-hydroxy-olean-12-en(13)-28-oic acid by several spectral and chemical analysis. this new triterpenoid saponin showed antimicrobial activity against various plants pathogenic bacteri ...200111841115
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
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