Publications

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 ...	2000	10899618
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 ...	2000	10905427
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 ...	2000	10905428
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 ...	2000	10913297
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 ...	2000	10919329
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 ...	2000	10930731
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 ...	2000	10939521
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 ...	2000	10945258
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. ...	2000	10947969
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 ...	2000	10952006
fungal morphology and fragmentation behavior in a fed-batch aspergillus oryzae fermentation at the production scale.	it 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 ...	2000	10992234
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 ...	2000	10993159
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.	2000	10993168
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 ...	2000	11015693
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 ...	2000	11024527
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.	2000	11026538
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 ...	2000	11040426
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 ...	2000	11042551
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 ...	2000	11055393
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 ...	2000	10620754
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 ...	1999	10628849
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 ...	1999	10628858
effects of agitation intensity on mycelial morphology and protein production in chemostat cultures of recombinant aspergillus oryzae.	the effects of agitation on fragmentation of a recombinant strain of aspergillus oryzae and its consequential effects on protein production have been investigated. constant mass, 5.3-l chemostat cultures at a dilution rate of 0.05 h-1 and a dissolved oxygen level of 75% air saturation, have been conducted at 550, 700, and 1000 rpm. these agitation speeds were chosen to cover a range of specific power inputs (2.2 to 12 kw m-3) from realistic industrial levels to much higher values. the use of a c ...	1999	10099554
on-line study of growth kinetics of single hyphae of aspergillus oryzae in a flow-through cell.	using image analysis the growth kinetics of the single hyphae of the filamentous fungus aspergillus oryzae has been determined on-line in a flow-through cell at different glucose concentrations in the range from 26 mg l-1 to 20 g l-1. the tip extension rate of the individual hyphae can be described with saturation type kinetics with respect to the length of the hyphae. the maximum tip extension rate is constant for all hyphae measured at the same glucose concentration, whereas the saturation con ...	1999	10099591
solid-state fermentation in rotating drum bioreactors: operating variables affect performance through their effects on transport phenomena.	aspergillus oryzae acm 4996 was grown on an artificial gel-based substrate and on steamed wheat bran during solid-state fermentations in 18.7 l rotating drum bioreactors. for gel fermentations fungal growth decreased as rotational speed increased, presumably due to increased shear. for wheat bran fermentations fungal growth improved under agitated compared to static culture conditions, due to superior heat and mass transfer. we conclude that the effects of operational variables on the performanc ...	1999	10099618
site-directed mutagenesis of a possible type 1 copper ligand of bilirubin oxidase; a met467gln mutant shows stellacyanin-like properties.	in our previous paper, we reported a mutant of recombinant myrothecium verrucaria bilirubin oxidase, in which the met467 residue was replaced by gly [shimizu, a. et al. (1999) biochemistry 38, 3034-3042]. this mutant displayed a remarkable reduction in enzymatic activity and an evident decrease in the intensity of the absorption band around 600 nm (type 1 charge transfer transition). in this study, we report the preparation of three met467 mutants (met467gln, met467his, and met467arg) and charac ...	1999	10101277
molecular cloning and expression of the novel fungal beta-glucosidase genes from humicola grisea and trichoderma reesei.	a novel fungal beta-glucosidase gene (bgl4) and its homologue (bgl2) were cloned from the cellulolytic fungi humicola grisea and trichoderma reesei, respectively. the deduced amino acid sequences of h. grisea bgl4 and t. reesei bgl2 comprise 476 and 466 amino acids, respectively, and share 73.1% identity. these beta-glucosidases show significant homology to plant beta-glucosidases belonging to the beta-glucosidase a (bga) family. both genes were expressed in aspergillus oryzae, and the recombina ...	1999	10101286
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 ...	1999	9990729
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 ...	1999	10023082
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 ...	1999	10029539
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 ...	1999	10052122
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 ...	1999	10052131
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 ...	1999	10052139
myrothecium verrucaria bilirubin oxidase and its mutants for potential copper ligands.	bilirubin oxidase (ec:1.3.3.5) 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 ...	1999	10074356
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 ...	1999	10091601
fermentation extract effects on the morphology and metabolism of the rumen fungus neocallimastix frontalis eb188.	the effects of aspergillus oryzae fermentation extract, amaferm, on the rumen fungus neocallimastix frontalis eb188 were studied. the secretion of cellulase was increased by 67% and rhyzoid development was increased 3.8-fold in the presence of extract. strength of fungal response increased in a dose-dependent manner and demonstrated a positive correlation between cell surface area and enzyme secretion. above certain concentrations of extract, however, the development of the fungus and enzyme sec ...	1999	10196744
targeted mutations in a trametes villosa laccase. axial perturbations of the t1 copper.	trametes villosa laccase was mutated on a tetrapeptide segment near the type 1 site. the mutations f463m and f463l were at the position corresponding to the type 1 copper axial methionine (m517) ligand in zucchini ascorbate oxidase. the mutations e460s and a461e were near the t1 copper site. the mutated trametes laccases were expressed in an aspergillus oryzae host and characterized. the e460s mutation failed to produce a transformant with meaningful expression. the f463l and a461e mutations did ...	1999	10212209
molecular and enzymic properties of recombinant 1, 2-alpha-mannosidase from aspergillus saitoi overexpressed in aspergillus oryzae cells.	for the construction of an overexpression system of the intracellular 1,2-alpha-mannosidase (ec 3.2.1.113) gene (msds) from aspergillus saitoi (now designated aspergillus phoenicis), the n-terminal signal sequence of the gene was replaced with that of the aspergillopepsin i (ec 3.4.23.18) gene (apns) signal, one of the same strains as described previously. then the fused 1, 2-alpha-mannosidase gene (f-msds) was inserted into the noti site between p-no8142 and t-agda in the plasmid pnan 8142 (9.5 ...	1999	10215597
identification of delta12-fatty acid desaturase from arachidonic acid-producing mortierella fungus by heterologous expression in the yeast saccharomyces cerevisiae and the fungus aspergillus oryzae.	based on the sequence information for the omega3-desaturase genes (from brassica napus and caenorhabditis elegans), which are involved in the desaturation of linoleic acid (delta9, delta12-18 : 2) to alpha-linolenic acid (delta9, delta12, delta15-18 : 3), a cdna was cloned from the filamentous fungal strain, mortierella alpina 1s-4, which is used industrially to produce arachidonic acid. homology analysis with protein databases revealed that the amino acid sequence showed 43.7% identity as the h ...	1999	10215899
antioxidant activity of a medicine based on aspergillus oryzae nk koji measured by a modified t-butyl peroxyl radical scavenging assay.	a koji-based medicine composed of powder of aspergillus oryzae nk koji, dried yeast, and lactobacilli koji had high antioxidant activity measured by a modified t-butyl peroxyl radical scavenging assay. this activity was mainly derived from a. oryzae nk koji. digestion of koji-making grain germ medium with several commercial enzymes also increased antioxidant activity. by two weeks of oral administration of a. oryzae nk koji, the serum lipid peroxide levels elevated in stz-induced diabetic rats c ...	1999	10227147
[oxidative injury and its defense system in vivo].	we and other researchers verified that excessively produced free radicals by neutrophils induce various diseases such as behçet's disease, mcls, sle (neutrophil-stimulated lymphocytes), ra (synovial fluid neutrophils), crohn's disease, colitis ulcerosa, and dermatitis herpetiformis (dühring). recently, it was reported that environmental toxic agents including herbicides such as paraquat, insecticides, nitrogen oxide, and ultraviolet radiation produce free radicals. nitrogen oxide, a main product ...	1999	10228384
stable high-copy-number integration of aspergillus oryzae alpha-amylase cdna in an industrial baker's yeast strain.	the aspergillus oryzae alpha-amylase cdna was placed under the control of the saccharomyces cerevisiae actin promoter (pact1) and introduced into the ribosomal dna locus of an industrial baker's yeast strain. to obtain a strain eligible for commercial use, we constructed an integrative cassette lacking bacterial dna sequences but containing the alpha-amylase cdna and ribosomal dna sequences to target the integration to this locus. high-copy-number integrants were obtained including a defective t ...	1999	10356263
a cross-sectional survey of sensitization to aspergillus oryzae-derived lactase in pharmaceutical workers.	the presence of ige-mediated occupational respiratory sensitization to microbial enzymes has been well documented in a variety of industries. aspergillus oryzae -derived lactase is used as a dietary aid for patients with lactose intolerance.	1999	10359899
production of recombinant der fi (a major mite allergen) by aspergillus oryzae.	der fi is a major mite allergen. to produce der fi by aspergillus oryzae, we placed a dna fragment encoding precursor-type recombinant der fi e(-1)k (reder fi e(-1) k), which had the c-terminal amino acid of the pro-sequence (glu) changed to lys, downstream of the glaa gene promoter and introduced it into aspergillus oryzae. in liquid culture, most of the reder fi e(-1)k produced by the transformants was degraded when culture was shaken vigorously. however, the degradation of reder fi e(-1)k was ...	1999	10361683
fermentation of low-salt miso as affected by supplementation with ethanol.	steam-cooked soybeans and rice koji were combined (1:1, w/w), mixed with 5% (w/w) nacl and ground into a fine paste. samples (30 g) were deposited in nylon/polyethylene plastic bags and supplemented with 10 ml of aqueous ethanol solutions to give concentrations of 0, 2.5, 5, 7.5, 10, 15, 20, and 25% ethanol. mixtures were homogenized, sealed, and incubated at 28 degrees c for eight weeks. mold populations were less than 3 log10 cfu/g in all miso products after four weeks of fermentation. yeast p ...	1999	10375131
molecular cloning and expression of the gene encoding a phospholipase a1 from aspergillus oryzae.	phospholipase a1 (pla1) is a hydrolytic enzyme that catalyzes removal of the acyl group from position 1 of lecithin to form lysolecithin. the genomic dna and cdna encoding pla1 from aspergillus oryzae were cloned with the mixed deoxyribonucleotide-primed polymerase chain reaction. the pla1 gene is composed of 1,056 bp and has four exons and three short introns (63, 54, and 51 bp). the deduced amino acid sequence of pla1 contained the n-terminal sequence of the mature pla1 analyzed by edman degra ...	1999	10380624
alcoholysis reactions from starch with alpha-amylases.	the ability of alpha-amylases from different sources to carry out reactions of alcoholysis was studied using methanol as substrate. it was found that while the enzymes from aspergillus niger and aspergillus oryzae, two well-studied saccharifying amylases, are capable of alcoholysis reactions, the classical bacterial liquefying alpha-amylases from bacillus licheniformis and bacillus stearothermophilus are not. the effect of starch and methanol concentration, temperature and ph on the synthesis of ...	1999	10386619
adapting protein solubility by glycosylation. n-glycosylation mutants of coprinus cinereus peroxidase in salt and organic solutions.	protein solubility is a fundamental parameter in biology and biotechnology. in the present study we have constructed and analyzed five mutants of coprinus cinereus peroxidase (cip) with 0, 1, 2, 4 and 6 n-glycosylation sites. all mutants contain man(x)(glcnac)(2) glycans. the peroxidase activity was the same for wild-type cip and all the glycosylation mutants when measured with the large substrate 2,2'-azino-bis(-3-ethylbenzthiazoline-6-sulfonic acid). the solubility of the five cip mutants show ...	1999	10407143
treatment of melasma using kojic acid in a gel containing hydroquinone and glycolic acid.	melasma is difficult to clear. many agents have been used, such as hydroquinone, and glycolic acid and glycolic acid peels, kojic acid, a tyrosinase inhibitor in the fungus aspergilline oryzae.	1999	10417583
purification, characterization, and heterologous expression in fusarium venenatum of a novel serine carboxypeptidase from aspergillus oryzae.	a novel serine carboxypeptidase (ec 3.4.16.1) was found in an aspergillus oryzae fermentation broth and was purified to homogeneity. this enzyme has a molecular weight of ca. 67,000, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and its specific activity is 21 u/mg for carbobenzoxy (z)-ala-glu at ph 4.5 and 25 degrees c. it has a ratio of bimolecular constants for z-ala-lys and z-ala-phe of 3.75. optimal enzyme activity occurs at ph 4 to 4.5 and 58 to 60 degrees c f ...	1999	10427010
immobilization of enzymes on spongy polyvinyl alcohol cryogels: the example of beta-galactosidase from aspergillus oryzae.	the catalytic properties of a beta-galactosidase from aspergillus oryzae, entrapped into a spongy polyvinyl alcohol cryogel, were studied. this polymeric matrix was selected because of its mild conditions of preparation and its stability, biocompatibility, structural strength and diffusive properties. the enzyme was entrapped, in high percentage, into cryogel sponges and its activity and kinetic parameters were determined and compared with those of the free enzyme, using as substrates o-nitrophe ...	1999	10434188
kinetics of alpha-amylase secretion in aspergillus oryzae.	pulse and pulse-chase experiments have been performed to study l-[(35)s] methionine incorporation and protein secretion kinetics in aspergillus oryzae. pulse experiments confirmed the mechanism of methionine uptake reported previously for penicillium chrysogenum (benko et al., 1967). pulse-chase experiments were carried out to investigate the alpha-amylase secretion kinetics in a. oryzae. no unglycosylated alpha-amylase was detected neither intracellularly nor extracellularly demonstrating that ...	1999	10440673
aspzincin, a family of metalloendopeptidases with a new zinc-binding motif. identification of new zinc-binding sites (his(128), his(132), and asp(164)) and three catalytically crucial residues (glu(129), asp(143), and tyr(106)) of deuterolysin from aspergillus oryzae by site-directed mutagenesis.	deuterolysin (ec 3.4.24.39; formerly designated as neutral proteinase ii) from aspergillus oryzae, which contains 1 g atom of zinc/mol of enzyme, is a single chain of 177 amino acid residues, includes three disulfide bonds, and has a molecular mass of 19,018 da. active-site determination of the recombinant enzyme expressed in escherichia coli was performed by site-directed mutagenesis. substitutions of his(128) and his(132) with arg, of glu(129) with gln or asp, of asp(143) with asn or glu, of a ...	1999	10446194
crystal structure of a ribonuclease from the seeds of bitter gourd (momordica charantia) at 1.75 a resolution.	the ribonuclease mc1 (rnase mc1) from seeds of bitter gourd (momordica charantia) consists of 190 amino acid residues with four disulfide bridges and belongs to the rnase t(2) family, including fungal rnases typified by rnase rh from rhizopus niveus and rnase t(2) from aspergillus oryzae. the crystal structure of rnase mc1 has been determined at 1.75 a resolution with an r-factor of 19.7% using the single isomorphous replacement method. rnase mc1 structurally belongs to the (alpha+beta) class of ...	1999	10446375
exposure-response relations of alpha-amylase sensitisation in british bakeries and flour mills.	to describe the levels of exposure to fungal alpha-amylase in british bakeries and flour mills, and to describe the relation between exposure to alpha-amylase and sensitisation to fungal alpha-amylase.	1999	10448329
the tama protein fused to a dna-binding domain can recruit area, the major nitrogen regulatory protein, to activate gene expression in aspergillus nidulans.	the area gene of aspergillus nidulans encodes a gata zinc finger transcription factor that activates the expression of a large number of genes subject to nitrogen metabolite repression. the amount and activity of the area protein under different nitrogen conditions is modulated by transcriptional, post-transcriptional, and post-translational controls. one of these controls of area activity has been proposed to involve the nmra protein interacting with the dna-binding domain and the extreme c ter ...	1999	10471703
identification of an nadh-cytochrome b(5) reductase gene from an arachidonic acid-producing fungus, mortierella alpina 1s-4, by sequencing of the encoding cdna and heterologous expression in a fungus, aspergillus oryzae.	based on the sequence information for bovine and yeast nadh-cytochrome b(5) reductases (cbrs), a dna fragment was cloned from mortierella alpina 1s-4 after pcr amplification. this fragment was used as a probe to isolate a cdna clone with an open reading frame encoding 298 amino acid residues which show marked sequence similarity to cbrs from other sources, such as yeast (saccharomyces cerevisiae), bovine, human, and rat cbrs. these results suggested that this cdna is a cbr gene. the results of a ...	1999	10473389
heterologous expression and product identification of colletotrichum lagenarium polyketide synthase encoded by the pks1 gene involved in melanin biosynthesis.	the colletotrichum lagenarium pks1 gene was expressed in the heterologous fungal host, aspergillus oryzae, under the starch-inducible alpha-amylase promoter to identify the direct product of polyketide synthase (pks) encoded by the pks1 gene. the main compound produced by an a. oryzae transformant was isolated and characterized to be 1,3,6,8-tetrahydroxynaphthalene (t4hn) as its tetraacetate. since the pks1 gene was cloned from c. lagenarium to complement the nonmelanizing albino mutant, t4hn wa ...	1999	10501004
new potent antioxidative o-dihydroxyisoflavones in fermented japanese soybean products.	a potent antioxidative 6-hydroxydaidzein (6-ohd) was newly isolated from soybean koji fermented with aspergillus oryzae. 6-ohd, in addition to 8-hydroxydaidzein and 8-hydroxygenistein, were found to be present in various fermented soybean products, including their koji. considering that these o-dihydroxyisoflavones had strong antioxidative activities, they may contribute to protecting from oxidative deterioration during the processing of fermented soybean products.	1999	10540753
molecular characterization of laccase genes from the basidiomycete coprinus cinereus and heterologous expression of the laccase lcc1.	a laccase from coprinus cinereus is active at alkaline ph, an essential property for some potential applications. we cloned and sequenced three laccase genes (lcc1, lcc2, and lcc3) from the ink cap basidiomycete c. cinereus. the lcc1 gene contained 7 introns, while both lcc2 and lcc3 contained 13 introns. the predicted mature proteins (lcc1 to lcc3) are 58 to 80% identical at the amino acid level. the predicted lcc1 contains a 23-amino-acid c-terminal extension rich in arginine and lysine, sugge ...	1999	10543807
cloning and characterization of the o-methyltransferase i gene (dmta) from aspergillus parasiticus associated with the conversions of demethylsterigmatocystin to sterigmatocystin and dihydrodemethylsterigmatocystin to dihydrosterigmatocystin in aflatoxin biosynthesis.	o-methyltransferase i catalyzes both the conversion of demethylsterigmatocystin to sterigmatocystin and the conversion of dihydrodemethylsterigmatocystin to dihydrosterigmatocystin during aflatoxin biosynthesis. in this study, both genomic cloning and cdna cloning of the gene encoding o-methyltransferase i were accomplished by using pcr strategies, such as conventional pcr based on the n-terminal amino acid sequence of the purified enzyme, 5' and 3' rapid amplification of cdna ends pcr, and ther ...	1999	10543813
effect of branch frequency in aspergillus oryzae on protein secretion and culture viscosity.	highly branched mutants of two strains of aspergillus oryzae (ifo4177, which produces alpha-amylase, and a transformant of ifo4177 [amg#13], which produces heterologous glucoamylase in addition to alpha-amylase) were generated by uv or nitrous acid mutagenesis. four mutants of the parental strain (ifo4177), which were 10 to 50% more branched than the parental strain, were studied in stirred batch culture and no differences were observed in either the amount or the rate of enzyme production. five ...	1999	10550770
a modified chemical procedure for rapid determination of glucosamine and its application for estimation of mold growth in peanut kernels and koji.	one-step hydrolysis of chitin to release glucosamine for quantitation was achieved by combining a chitin-containing sample (10-200 mg of sample size) in a test tube with 1 ml of 10 m hcl followed by vacuum treatment for 10 min, incubation at 28 degrees c for 30 min, replenishment with 3 ml of deionized water, nitrogen flushing, screw capping, and heat treatment at 140 degrees c for 60 min. a phosphate buffer solution (ph 12.5, 0.2 m) was effective in ph stabilization and enhancing colorimetric d ...	1999	10552485
characteristics and use of okara, the soybean residue from soy milk production--a review.	large quantities of okara produced annually pose a significant disposal problem. it contains mostly crude fiber composed of cellulose, hemicellulose, and lignin, about 25% protein, 10-15% oil, but little starch or simple carbohydrates. it is a suitable dietary additive in biscuits and snacks because it reduces calorie intake and increases dietary fiber. the high-quality protein fraction has good water holding and emulsifying qualities and contains a peptide with anti-hypertension effects. the pe ...	1999	10563901
delta6-fatty acid desaturase from an arachidonic acid-producing mortierella fungus. gene cloning and its heterologous expression in a fungus, aspergillus.	a dna fragment was cloned from the fungal strain, mortierella alpina 1s-4 (which is used industrially to produce arachidonic acid), after pcr amplification with oligonucleotide primers designed based on the sequence information for delta6-desaturase genes (from borage and caenorhabditis elegans), which are involved in the desaturation of linoleic acid (delta9, delta12-18:2) to gamma-linolenic acid (delta6, delta9, delta12-18:3). this fragment was used as a probe to isolate a cdna clone with an o ...	1999	10570972
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.	1999	10586505
identification of enzymes and quantification of metabolic fluxes in the wild type and in a recombinant aspergillus oryzae strain	two 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 ...	1999	9872753
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 ...	1999	9872754
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.	1999	9872797
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 ...	1999	9884411
production of alkaline protease by a genetically engineered aspergillus oryzae u1521.	the production of alkaline protease of aspergillus oryzae u1521 was examined in liquid culture. in a culture of defatted soybean only, it gave satisfactory enzyme yields at 584,000 u/g defatted soybean. when various carbohydrates were supplemented, enzyme production was significantly increased. an increase in production by lactose was the most marked. enrichment with casitone or casein increased productivity, but not cornsteep solid. media formulation (g/l) of defatted soybean 10, lactose 5, cas ...	1999	12501379
[quantitative determination of 3-nitropropionic acid by hplc].	a reversed-phase liquid chromatographic method to determine the concentration of 3-nitropropionic acid (3-npa) in the culture medium with aspergillus oryzae was developed. the culture medium samples were extracted with ethyl acetate. the analytic column was hypersil c18 and the mobile phase was ch3cn/kh2po4(0.02 mol/l, ph 3.0) (1/3, v/v). the sample was detected at uv210 nm. the detection limit of this method was at 10 micrograms/kg level and recoveries were 90.1%-98.0.	1999	12712703
cloning and functional analysis of the aspergillus oryzae conidiation regulator gene brla by its disruption and misscheduled expression.	we cloned the brla gene from aspergillus oryzae genomic dna using the a. nidulans brla gene as a probe. a 3.1-kb ecori-bali genomic dna fragment was cloned and sequenced. the deduced amino acid sequence revealed 70% identity with a. nidulans brla and contained two c2h2 zinc finger motifs in its carboxyl terminus, and the promoter sequence contained a 43-bp highly conserved region, indicating that the cloned gene was an a. oryzae homologue of a. nidulans brla. disruption of the brla gene by homol ...	1999	16232494
properties of the peptides liberated from rice protein in sokujo-moto.	in the supernatant of sokujo-moto, a high level of acid carboxypeptidase (acp) activity and a large amount of peptides were observed, however, the amount of free amino acids liberated was small. in order to determine why these peptides were not hydrolyzed to any significant degree by the acp, the properties of the peptides in sokujo-moto were investigated in this study. peptides were fractionated from sokujo-moto by ion exchange column chromatography. acp purified from rice-koji (rice overgrown ...	1999	16232611
aspergillus oryzae palbory encodes a calpain-like protease: homology to emericella nidulans palb and conservation of functional regions.	we have cloned and sequenced genomic dna of aspergillus oryzae palbory, orthologue of emericella nidulans palb, which encodes a calpain-like protease modulating a signal transduction pathway during alkaline adaptation. the deduced amino acid sequence of palbory is 70.0% identical to palb over its entire length. the regions with high similarity revealed possible domains important for their function. this is the first step towards understanding the alkaline adaptation mechanism of a. oryzae, which ...	1999	16232641
cloning and characterization of the bipa gene encoding er chaperone bip from aspergillus oryzae.	we describe the cloning and characterization of the er localized chaperone (bip) encoding gene from the filamentous fungus aspergillus oryzae. the bip encoding gene, designated bipa, has three introns and encodes a protein with 672 amino acids, which has a high homology with various bip-like proteins. sequences resembling heat shock elements (hses) and unfolded protein response (upr) elements, as found in the saccharomyces cerevisiae kar2 promoter, are present in the 5' no coding region of the b ...	1999	16232647
production and some properties of salt-tolerant beta-xylosidases from a shoyu koji mold, aspergillus oryzae in solid and liquid cultures.	beta-xylosidase production from a shoyu (soy sauce) koji mold, aspergillus oryzae hl15, cultured in solid and liquid media was examined and some properties of the enzymes were studied. three beta-xylosidases (xy11, xy12 and xy13) were easily extracted with 0.5% nacl from a solid medium and purified homogeneously on sds-page by chromatography. on the other hand, in a liquid medium, a. oryzae hl15 produced mainly cell-wall-bound beta-xylosidases which could not be extracted with 0.5% nacl or any d ...	1999	16232648
dry fermented sausages made with a protease from aspergillus oryzae and/or a starter culture.	sausages with a starter culture (lactobacillus plantarum + staphylococcus carnosus), a protease (fungal protease from solvay enzymes) and both enzyme/starter were produced from the same raw matter under the same conditions for 15 days of ripening. the lowest ph values were found in fungal/starter sausages. significantly higher amounts of free amino acids (faa) were seen in sausages containing enzyme from the 3rd day of ripening, whereas amino acids from peptides (paa) were significantly higher o ...	1999	22062704
purification and characterization of isoamyl alcohol oxidase ("mureka"-forming enzyme).	isoamyl alcohol oxidase (iaaod) was purified to apparent homogeneity on sds-page from ultrafiltration (uf) concentrated sake. iaaod was a glycoprotein, a monomeric protein with an apparent molecular mass of 73 and 87 kda, by sds-page and gel filtration on hplc, respectively. iaaod showed high substrate specificity toward c5 branched-chain alkyl alcohol (isoamyl alcohol), and no activity toward shorter (c1-c4) or longer (c7-c10) alkyl alcohols tested. iaaod was stable between ph 3.0-6.0 at 25°c. ...	1999	27380232
cysteine protease inhibitors produced by the industrial koji mold, aspergillus oryzae o-1018.	aspergillus oryzae o-1018 (ferm p-15834) separated from industrial koji for brewing sake was found to produce five papain-inhibitory compounds in the culture supernatant. the five isolated inhibitors were named cpi-1 to cpi-5, and their structures were elucidated by spectroscopic analyses and chemical degradation. we determined the structures of cpi-2, cpi-3 and cpi-4 as 4-amino-1-[[n- [(2s, 3s)-3-trans-carboxyoxiran-2-carbonyl]-l-isoleucyl] amino]butane, 5-amino-1-[[n-[(2s, 3s)-3-trans-carboxyo ...	1998	27392587
transglycosylation of thiamin by fungal β-n-acetylhexosaminidases.	a new thiamin glycoside, e.g., thiamin β-d-2-deoxy-2-acetamidoglucopyranoside was prepared by transglycosylation by β-n-acetylhexosaminidase from aspergillus oryzae and characterized spectrally. series of other fungal β-n-acetyl-hexosaminidases from a. awamori, a. tamari, a. terreus, and penicillium oxalicum were shown to be able to synthesize the same compound in lower yields.	1998	27392399
preliminary studies on production of extracellular lipase by an isolate from soil : aspergillus flavus oryzae.	in our routine screening programme, using agar diffusion assay method, lipolytic activity was detected around a colony of a fungus. the fungus was isolated from a soil sample which was brought from a location near oil-mill. this lipolytic fungus was then identified to belong to aspergillus flavus oryzae. a medium was then formulated and optimized which would not only support good growth but also would yield good extracellular lipolytic activity. it was observed that a conventional carbohydrate-p ...	1998	16961201
evolutionary relationships among aspergillus oryzae and related species based on the sequences of 18s rrna genes and internal transcribed spacers.		1998	12501432
chaperonin groe-facilitated refolding of disulfide-bonded and reduced taka-amylase a from aspergillus oryzae.	the refolding characteristics of taka-amylase a (taa) from aspergillus oryzae in the presence of the chaperonin groe were studied in terms of activity and fluorescence. disulfide-bonded (intact) taa and non-disulfide-bonded (reduced) taa were unfolded in guanidine hydrochloride and refolded by dilution into buffer containing groe. the intermediates of both intact and reduced enzymes were trapped by groel in the absence of nucleotide. upon addition of nucleotides such as atp, adp, ctp or utp, the ...	1998	9930680
localization of a phosphatidylglycerol/phosphatidylinositol transfer protein in aspergillus oryzae.	the subcellular localization of the phosphatidylglycerol/phosphatidylinositol transfer protein (pg/pi-tp) of aspergillus oryzae was investigated using western blot analysis of the cell protein extracts, a cellular membrane fractionation technique, and transmission electron microscopy. the pg/pi-tp, as detected by western blot analysis with a specific immune serum, was found to be mainly cytoplasmic and partly associated with intracellular membranes. a fractionation experiment was conducted after ...	1998	9933913
action of trichoderma reesei and aspergillus oryzae esterases in the deacetylation of hemicelluloses.	xylans and mannans contain different esterified substituents such as acetyl, feruloyl and p-coumaroyl side groups. the functions of hemicellulose-deacetylating esterases of trichoderma reesei and aspergillus oryzae are discussed in this paper. both fungi produce multiple esterases and two different esterases were isolated from both t. reesei and a. oryzae. the enzymes differed significantly in their substrate specificities. acetyl xylan esterase of t. reesei was highly active on polymeric xylan ...	1998	9477552
construction of a low-serine-type-carboxypeptidase-producing mutant of aspergillus oryzae by the expression of antisense rna and its use as a host for heterologous protein secretion.	using an antisense control strategy, we isolated an aspergillus oryzae mutant that produced low levels of carboxypeptidases (cpases). the mutant tfc-1 expressed the antisense rna of the structural gene of cpase o and showed about 30% of the cpase activity in the parent strain. gel filtration analysis indicated that this mutant decreased the cpase activities not only of cpase o but also of cpase o-1 and o-2. this result indicated that the antisense rna was able to control the expression of the cp ...	1998	9487708
nucleotide sequence of an alternative glucoamylase-encoding gene (glab) expressed in solid-state culture of aspergillus oryzae.	the dna (glab) and a cdna-encoding glucoamylase produced in solid-state culture of aspergillus oryzae were cloned using oligodeoxyribonucleotide probes derived from internal amino acid sequences of the enzyme. comparison of the nucleotide sequences of a genomic dna fragment with its cdna showed the glab gene carried three exons interrupted by two introns and had an open reading frame encoding 493 aa residues. the 5'-flanking region had a tata box at nt -87 from the start codon and two putative c ...	1998	9511753
overproduction of 1,2-alpha-mannosidase, a glycochain processing enzyme, by aspergillus oryzae.	a recombinant strain of aspergillus oryzae has been constructed in which 1,2-alpha-mannosidase, an intracellular glycochain processing enzyme with specificity toward 1,2-alpha-mannosidic linkages, has been overexpressed. for the construction, the n-terminal signal-encoding sequence of the 1,2-alpha-mannosidase gene (msdc) from penicillium citrinum was replaced with that of the aspergillopepsin i signal, and the fused gene was inserted between amyb promoter-terminator elements in the expression p ...	1998	9532788
hydrolysis of a slow cyclic thiophosphate substrate of rnase t1 analyzed by time-resolved crystallography.	here we present a time-resolved crystallographic analysis of the hydrolysis of exo (sp) guanosine 2',3'-cyclophosphorothioate by rnase t1. the use of a slow substrate and fast crystallization methods made it possible to perform the study with conventional data-collection techniques. the results support the idea that the hydrolysis reaction proceeds through a mechanism that is the inverse of the transesterification reaction. in addition, the structures provide an explanation for the differential ...	1998	9546218
characterization of recombinant rhamnogalacturonan alpha-l-rhamnopyranosyl-(1,4)-alpha-d-galactopyranosyluronide lyase from aspergillus aculeatus. an enzyme that fragments rhamnogalacturonan i regions of pectin.	the four major oligomeric reaction products from saponified modified hairy regions (mhr-s) from apple, produced by recombinant rhamnogalacturonan (rg) alpha-l-rhamnopyranosyl-(1, 4)-alpha-d-galactopyranosyluronide lyase (rrg-lyase) from aspergillus aculeatus, were isolated and characterized by 1h-nuclear magnetic resonance spectroscopy. they contain an alternating rg backbone with a degree of polymerization of 4, 6, 8, and 10 and with an alpha-delta-(4,5)-unsaturated d-galactopyranosyluronic aci ...	1998	9576783
[regulation of gene expression in aspergilli].	the aspergillus oryzae taka-amylase a (taa) gene has been used as a model gene to characterize the regulatory mechanisms of gene expression in aspergilli. taa gene contained a typical eukaryotic promoter with a tata box and putative regulatory elements such as a ccaat sequence in its 5'-noncoding region. a nuclear protein designated ancp bound to the ccaat sequence. replacement of the ccaat sequence with cgtaa was found to abolish the binding of ancp and to have an inhibitory effect on taa promo ...	1998	9580033
purification and characterization of triacylglycerol lipase from aspergillus oryzae.	triacylglycerol lipase (l3) was purified from aspergillus oryzae rib128 by ammonium sulfate fractionation, acetone precipitation, anion-exchange chromatography, and gel filtration. the purified enzyme was formed from a glycoprotein and a monomeric protein with molecular masses of 25 and 29 kda, by sds-page and gel filtration, respectively. the optimum ph at 40 degrees c was 5.5 and the optimum temperature at ph 5.5 was 40 degrees c. the enzyme was stable between a ph range of 4.0-7.5 at 30 degre ...	1998	9614707
xylooligosaccharide production by aspergillus oryzae 13 immobilized on a nonwoven fabric.	immobilized mycelia were screened for xylooligosaccharide production from xylan, and 20 strains of aspergillus oryzae were selected. for its high activity and operational stability of xylooligosaccharides formation, immobilized a. oryzae 13 was selected for further examination. batch production of xylooligosaccharides from xylan by the immobilized mycelia was repeated a total of 4 times.	1998	9614714
increase in milk yield of commercial dairy herds fed a microbial and enzyme supplement.	a microbial and enzyme supplement fed at 21.2 g/d per cow to 46 virginia dairy herds increased the milk yield of 31 herds (17 significantly) and decreased the milk yield of 15 herds (7 significantly). effects of season were important but consistent with overall results. herds began receiving the supplement, which contained dried fermentation products of aspergillus oryzae, bacillus subtilis, lactobacillus acidophilus, and yeast culture, midway between the first and second monthly dairy herd impr ...	1998	9621238
alkaline serine proteinase: a major allergen of aspergillus oryzae and its cross-reactivity with penicillium citrinum.	aspergillus species are common indoor airborne fungi and have been considered as causative agents of human allergic disorders. however, allergens of different aspergillus species have not been effectively characterized. the object of this study was to identify and characterize ige-binding components of aspergillus oryzae.	1998	9623506
polygalacturonase gene expression in ripe melon fruit supports a role for polygalacturonase in ripening-associated pectin disassembly.	ripening-associated pectin disassembly in melon is characterized by a decrease in molecular mass and an increase in the solubilization of polyuronide, modifications that in other fruit have been attributed to the activity of polygalacturonase (pg). although it has been reported that pg activity is absent during melon fruit ripening, a mechanism for pg-independent pectin disassembly has not been positively identified. here we provide evidence that pectin disassembly in melon (cucumis melo) may be ...	1998	9625689
ben1 and zen1 cdnas encoding s1-type dnases that are associated with programmed cell death in plants.	we describe the isolation and characterization of cdnas encoding two dnases that may be involved in the programmed death of plant cells: a 35-kda nuclease of barley, which had been reported to be secreted from the aleurone layer into the endosperm during germination and may be responsible for the digestion of nuclear dna in the course of degradation (cell death) of endosperm, and a 43-kda nuclease of zinnia, which appears transiently in association with differentiation to tracheary elements and ...	1998	9650576
effect of process conditions on recovery of protein activity after freezing and freeze-drying.	the objective of this research was to gain a better understanding of the degree to which recovery of activity of model proteins after freeze-drying can be maximized by manipulation of freeze-dry process conditions in the absence of protective solutes. catalase, beta-galactosidase and lactate dehydrogenase (ldh) were used as model proteins. all of the three proteins exhibited a concentration-dependent loss of activity after freezing, with significantly higher recovery at higher concentration. the ...	1998	9653629
effect of n-linked glycosylation on secretion, activity, and stability of alpha-amylase from aspergillus oryzae.	the effect of n-linked glycosylation on secretion, activity, and stability of alpha-amylase from aspergillus oryzae grown as dispersed filaments was studied. in the presence of tunicamycin the fungus grew either as dispersed filaments or as one large pellet, whereas growth was as dispersed filaments in all control cultures. the presence of tunicamycin affected neither biomass, level of secreted alpha-amylase, nor total amount of secreted protein in cultures growing as dispersed filaments. in the ...	1998	9662611