Publications

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in vivo antioxidant activity of okara koji, a fermented okara, by aspergillus oryzae.the antioxidants in okara koji (ok), an okara (oc) fermented by aspergillus oryzae, gamma-tocopherol, delta-tocopherol, genistin, daizein, genistein, and 3-hydroxyanthranilic acid were identified by hplc. ok's extract with 80% methanol strongly inhibited linoleate peroxidation, much more than other ok's extracts with hexane or hot water. the methanol extract accelerated 12-hydroxyeicosatetraenoic acid formation in membrane lipids at 10(-3) concentration, but inhibited the formation at higher con ...19979438976
characteristics of antibody responses induced in mice by protein allergens.whereas many foreign proteins are immunogenic, only a proportion is also allergenic, having the capacity to induce the quality of immune response necessary to support the production of ige antibody. we have demonstrated previously that intraperitoneal administration to mice of proteins such as ovalbumin (ova) or the industrial enzyme a. oryzae lipase, which possess significant allergenic potential, stimulates the production of both igg and ige antibody. identical exposure to bovine serum albumin ...19979449226
trisaccharide synthesis by glycosyl transfer from p-nitrophenyl beta-d-n-acetylgalactosaminide on to disaccharide acceptors catalysed by the beta-n-acetylhexosaminidase from aspergillus oryzae.the beta-n-acetylhexosaminidase from aspergillus oryzae catalysed the transfer of beta-d-n-acetylgalactosaminyl residues from p-nitrophenyl beta-d-n-acetylglucosaminide on to disaccharide acceptors consisting of thioethyl glycosides of alpha-d-glc-(1-->4)-beta-d-glc, beta-d-glc-(1-->4)-beta-d-glc and beta-d-glc-(1-->6)-beta-d-glc. the principle of 'anomeric control' was exemplified by the results which showed that an alpha-linkage between the units of the acceptor favoured exclusively the format ...19979648256
regioselectivity of the enzymatic transgalactosidation of d- and l-xylose catalysed by beta-galactosidases.the regioselectivity of enzymatic transgalactosidation depends on the source of the beta-galactosidase used. when the galactosyl acceptor only contains secondary hydroxyl groups, e.g., d- or l-xylose, it is possible to find an enzyme that catalyses preferentially the synthesis of any of the three regioisomers 4-, 3- and 2-o-beta-d-galactopyranosyl-d-xylose (1, 2 and 3, respectively) or 4-, 3- and 2-o-beta-d-galactopyranosyl-l-xylose (4, 5 and 6, respectively). enriched mixtures in 1, 2 or 3 were ...19979648257
development of a two-site enzyme-linked immunosorbent assay for alpha-amylase from aspergillus oryzae based on monoclonal antibodies.a two-site monoclonal antibody elisa was developed to quantify the allergen asp o 2 (alpha-amylase from aspergillus oryzae). two mabs recognizing distinct epitopes were selected, enriched by in vitro production in a modular minifermenter and affinity-purified. the first antibody was bound to microtiter plates which were then incubated with samples containing the allergen. bound allergen was detected using a biotinylated second antibody and peroxidase-polymer-labelled streptavidin. the assay had ...19979502588
effect of organic solvents on enantioselectivity of protease catalysis.the protease-catalyzed transesterifications between n-trifluoroacetyl-dl-phenylalanine 2,2,2-trifluoroethyl ester and 1-propanol were studied in a variety of anhydrous organic solvents at 30 degrees c. the protease preparations lyophilized from phosphate buffer solutions (ph 8.0) were used as catalysts. the organic solvent affected both rate of reaction and enantioselectivity differently. proteases such as aspergillus oryzae protease, subtilisin carlsberg, and subtilisin bpn' always preferred th ...199718629956
proteolysis in dry fermented sausages: the effect of selected exogenous proteases.the effect of three commercial proteases (pronase e from streptomyces griseus, aspartyl proteinase from aspergillus oryzae and papain) on protein breakdown and the sensory characteristics of dry fermented sausages was investigated. water soluble, non-protein, 5% phosphotungstic acid soluble, 5% sulphosalicylic acid soluble and total volatile basic nitrogen contents increased during fermentation, stabilizing later until the end of ripening (26th day). nitrogen values were always greater in the as ...199722061850
isolation of taka-amylase a peptides required for substrate binding.an α-amylase from aspergillus oryzae, taka-amylase a (taa), was cleaved into peptide fragments by an acid protease. inactivation of taa was greatly retarded by the addition of α-cyclodextrin or ca(2+). taa peptide fragments were separated into two groups having no and high affinity to the substrate, soluble starch. this separation was done by the forced affinity chromatography method by a column of epichlorohydrin cross-linked soluble starch gel. three peptides were isolated from the high-affini ...199727396736
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 ...19989511753
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 ...19989532788
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 ...19989546218
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 ...19989576783
[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 ...19989580033
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 ...19989614707
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.19989614714
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 ...19989621238
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.19989623506
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 ...19989625689
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 ...19989930680
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 ...19989933913
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 ...19989650576
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 ...19989653629
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 ...19989662611
effect of aspergillus oryzae extract alone or in combination with antimicrobial compounds on ruminal bacteria.the effect of an aspergillus oryzae fermentation extract on the growth rates of pure cultures of ruminal bacteria was determined. bacteria were grown in an anaerobic ruminal fluid and carbohydrate medium. a sterile filtrate made with 10% a. oryzae was added to the medium at 2 or 5% (vol/vol) to provide a final a. oryzae concentration of 2 or 5 mg/ml, respectively. the filtrate had no effect on the growth rates of 10 of the 19 ruminal bacteria tested; however, the filtrate increased the growth ra ...19989684165
production of amylases from rice by solid-state fermentation in a gas-solid spouted-bed bioreactora gas-solid spouted-bed bioreactor was developed to produce amylases from rice in solid-state fermentation by aspergillus oryzae. the spouted-bed bioreactor was developed to overcome many of the problems inherent to large-scale solid-state fermentation, including mass- and heat-transfer limitations in the conventional tray reactors and solids-handling difficulties seen in packed-bed bioreactors. the solid-state fermentation results from the tray-type reactor with surface aeration were poor becau ...19989694679
utilization of the tef1-alpha gene (tef1) promoter for expression of polygalacturonase genes, pgaa and pgab, in aspergillus oryzae.for the development of an efficient gene expression system in a shoyu koji mold aspergillus oryzae kbn616, the tef1 gene, encoding translation-elongation factor 1 alpha, was cloned from the same strain and used for expression of polygalacturonase genes. the tef1 gene comprised 1647 bp with three introns. the tef1-alpha protein consisted of 460 amino acids possessing high identify to other fungal tef proteins. two nucleotide sequences homologous to the upstream activation sequence, characterized ...19989720204
aspergillus oryzae with and without a homolog of aflatoxin biosynthetic gene ver-1.part of the nucleotide sequence of the ver-1 homolog in each of two strains of aspergillus oryzae, aspergillus sojae, and aspergillus flavus were compared with two homologs in aspergillus parasiticus. the homologs in a. oryzae and a. sojae (non-aflatoxin-producers) exhibited an extremely high degree (93.8-99.8% for a. oryzae, and 96.0-99.5% for a. sojae) of sequence identify with that of a. flavus and a. parasiticus. no two strains within the same species, except a. sojae, were identical. no seq ...19989720206
recognition of single-stranded dna by nuclease p1: high resolution crystal structures of complexes with substrate analogs.the reaction mechanism of nuclease p1 from penicillium citrinum has been investigated using single-stranded dithiophosphorylated di-, tetra-, and hexanucleotides as substrate analogs. the complexes crystallize in tetragonal and orthorhombic space groups and have been solved by molecular replacement. the high resolution structures give a clear picture of base recognition by p1 nuclease at its two nucleotide-binding sites, especially the 1.8 a structure of a p1-tetranucleotide complex which can be ...19989726413
role of the regulatory gene area of aspergillus oryzae in nitrogen metabolism.the area gene of aspergillus oryzae was cloned by cross-hybridization with the aspergillus nidulans area gene and was found to encode an 866-amino-acid protein that is very similar to other fungal nitrogen regulatory proteins. the a. oryzae area gene can complement a. nidulans area loss-of-function mutations. functional analyses indicated that the n-terminal region of the a. oryzae area protein was dispensable for function and revealed a probable acidic activation domain in the protein. c-termin ...19989726865
isolation of the gene and characterization of the enzymatic properties of a major exoglucanase of humicola grisea without a cellulose-binding domain.an exoglucanase gene was cloned from a cellulolytic fungus, humicola grisea. dna sequencing of this gene, designated as exo1, revealed that it contained four introns in the coding region. the deduced amino acid sequence of exo1 was 451 amino acids in length and showed 57.7% identity with that of h. grisea cellobiohydrolase 1 (cbh1), but lacked the typical domain structures of a cellulose-binding domain and a hinge region. transcriptional analysis of the exo1 and cbh1 genes showed that the expres ...19989756616
purification, characterization, and substrate specificity of a novel highly glucose-tolerant beta-glucosidase from aspergillus oryzae.aspergillus oryzae was found to secrete two distinct beta-glucosidases when it was grown in liquid culture on various substrates. the major form had a molecular mass of 130 kda and was highly inhibited by glucose. the minor form, which was induced most effectively on quercetin (3,3',4',5,7-pentahydroxyflavone)-rich medium, represented no more than 18% of total beta-glucosidase activity but exhibited a high tolerance to glucose inhibition. this highly glucose-tolerant beta-glucosidase (designated ...19989758774
regulation of aflr and its product, aflr, associated with aflatoxin biosynthesis.we studied the role of the regulatory gene aflr and its product, aflr, in the biosynthesis of aflatoxin in aspergillus. western blot and enzyme-linked immunosorbent assay analyses revealed that aflatoxin b1 accumulation was directly related to aflr expression and was regulated by various environmental and nutritional conditions, including temperature, air supply, carbon source, nitrogen source, and zinc availability. expression of an aflatoxin biosynthetic pathway structural gene, omta, was regu ...19989758790
[quantification of inhaled exposure to alpha-amylase in 2 bakeries].baker's asthma and baker's rhinitis are among the most frequent occupational diseases. a major cause is the high exposure to flour dust in the workplace and to allergenic enzymes like alpha-amylase from aspergillus oryzae (allergen name: asp o 2).19989774870
the effect of saccharomyces cerevisiae and aspergillus oryzae on the digestion of the cell wall fraction of a mixed diet in defaunated and refaunated sheep rumen.the objective of this study was to determine the effect of two probiotics, saccharomyces cerevisiae (sc) and aspergillus oryzae (ao), without their culture medium, on the digestion of plant cell wall components in sheep that had been successively defaunated and refaunated. six sheep fitted with large rumen cannulae were used to study 1) defaunated sheep with no probiotic, 2) defaunated sheep with sc or ao, 3) refaunated sheep with no probiotic, 4) refaunated sheep with sc or ao. the apparent dig ...19989795984
overproduction of recombinant trichoderma reesei cellulases by aspergillus oryzae and their enzymatic properties.we have established an expression system of trichoderma reesei cellulase genes using aspergillus oryzae as a host. in this system, the expression of t. reesei cellulase genes were regulated under the control of a. oryzae taka-amylase promoter and the cellulase genes were highly expressed when maltose was used as a main carbon source for inducer. the production of recombinant cellulases by a. oryzae transformants reached a maximum after 3-4 days of cultivation. in some cases, proteolysis of recom ...19989828459
microorganism inactivation using high-pressure generation in sealed vessels under sub-zero temperature.in order to test the possibility of utilizing high pressure in bioscience and biotechnology, a simple method for high-pressure generation and its use for microbial inactivation have been studied. when a pressure vessel was filled with water, sealed tightly and cooled to sub-zero temperatures, high pressure was generated in the vessel. the pressure generation was 60 mpa at -5 degrees c, 103 mpa at -10 degrees c, and 140 mpa at -15 degrees c, -20 degrees c, and -22 degrees c. the high pressure gen ...19989830091
improvement of promoter activity by the introduction of multiple copies of the conserved region iii sequence, involved in the efficient expression of aspergillus oryzae amylase-encoding genes.the role of the conserved sequence region iii in the promoter regions of the amylase-encoding genes amyb, glaa and agda of aspergillus oryzae was examined. introduction of multiple copies of the region iii fragment into the agda promoter resulted in a significant increase in promoter activity at the transcriptional level. this result suggests that the fragment comprising region iii consists of one or more cis-acting sequence(s). moreover, expression of the agda gene under the control of the impr ...19989830097
characterization of the prolyl dipeptidyl peptidase gene (dppiv) from the koji mold aspergillus oryzae.the koji mold aspergillus oryzae secretes a prolyl dipeptidyl peptidase (dppiv) when the fungus is cultivated in a medium containing wheat gluten as the sole nitrogen and carbon source (mmwg). we cloned and sequenced the dppiv gene from an a. oryzae library by using the a. fumigatus dppiv gene as a probe. reverse transcriptase pcr experiments showed that the a. oryzae dppiv gene consists of two exons, the first of which is only 6 bp long. the gene encodes an 87.2-kda polypeptide chain which is s ...19989835566
biodegradation of ornithine-containing polylysine hydrogels.the degradation of the cross-linked cationic poly(amino acid)-glutaraldehyde (ga) hydrogels by two kinds of proteolytic enzymes, trypsin and aspergillus protease type xxiii, and by seven species of soil filamentous fungi has been investigated using homo- and copolypeptides of lysine (lys) and ornithine (orn). trypsin degraded the hydrogels prepared from poly(lys) and copoly(lys orn)s but not poly(orn), while aspergillus protease degraded all of them. degradation time of hydrogels by the two prot ...19989855186
cloning of a thermostable ascorbate oxidase gene from acremonium sp. hi-25 and modification of the azide sensitivity of the enzyme by site-directed mutagenesis.a gene encoding a thermostable ascorbate oxidase (asom) was cloned from acremonium sp. hi-25 and sequenced. the gene comprised 1709 bp and was interrupted by a single intron of 57 bp. asom consisted of 551 amino acids including a signal peptide with a molecular mass of 61200, and contained four histidine-rich regions with high sequence homology to the corresponding regions of other multicopper oxidases. the asom gene was expressed in aspergillus nidulans under the aspergillus oryzae taka-amylase ...19989858779
repression of the expression of genes encoding xylanolytic enzymes in aspergillus oryzae by introduction of multiple copies of the xynf1 promoter.a xylanase gene, xynf1, was cloned and characterized from a shoyu koji mould aspergillus oryzae kbn616. the xynf1 gene was found to be comprised of 1484 bp with ten introns. the deduced amino acid sequence encodes a protein consisting of 327 amino acids (35,402 da) which is very similar to the fungal family f xylanases such as aspergillus nidulans xlnc, aspergillus kawachii xyna and penicillium chrysogenum xylp. the intron/exon organization of xynf1 is very similar to that of the fungal family f ...19989866173
transcript of a homolog of aflr, a regulatory gene for aflatoxin synthesis in aspergillus parasiticus, was not detected in aspergillus oryzae strains.some strains of aspergillus oryzae were shown to have homologs of aflr, a regulatory gene for aflatoxin synthesis in aspergillus parasiticus. transcription of an aflr homolog was examined in six strains of a. oryzae having the homologs, using polymerase chain reaction (pcr) coupled with reverse transcription. no pcr product was obtained when the rna prepared from the a. oryzae strains cultivated under aflatoxin-producing condition was used as template for amplification of the aflr cdna. by contr ...19989868774
hydrolase-catalyzed beta-mannosylations.transmannosylations catalysed by beta-mannosidase from snail viscera or beta-galactosidase from aspergillus oryzae were accomplished with 4-nitrophenyl beta-d-mannopyranoside as donor substrate. with suitable hydrophobic acceptor molecules preferentially beta1-4-linked disaccharides were obtained. the activities of both glycosidases in buffer cosolvent mixtures were determined, and conditions for their immobilization were elaborated and optimized. a model of the enzymic transfer mechanism is sug ...19989870351
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 ...19989477552
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 ...19989487708
successful treatment of severe atopic dermatitis-complicated cataract and male infertility with a natural product antioxidant.there has been a recent dramatic change in the features of atopic dermatitis and male infertility, including a marked increased prevalence of severe and treatment-resistant atopic dermatitis; an increase in severe atopic dermatitis complicated by cataracts, especially in urban and industrial areas; and an increase in the number of infertile men with poor sperm motility. previously we have attributed these changes to the increased free radicals produced by environmental toxicity. we have reported ...19989638503
growth and product formation of aspergillus oryzae during submerged cultivations: verification of a morphologically structured model using fluorescent probes.a morphologically structured model is well suited for obtaining a good description of growth and product formation of filamentous fungi for use in a process model, for example. this article describes a new morphologically structured model and its application to an alpha-amylase producing strain of aspergillus oryzae. the model is based on a division of the fungal hyphae into three different regions: an extension zone, representing the tips of the hyphae; an active region, which is responsible fo ...199810099209
on-line study of fungal morphology during submerged growth in a small flow-through cell.a flow-through cell is designed to measure the growth kinetics of hyphae of aspergillus oryzae grown submerged in a well controlled environment. the different stages of the growth process are characterized, from the spore to the fully developed hyphal element with up to 60 branches and a total length lt up to 10,000 micrometer. spore swelling is found to occur without change in the form of the spore (circularity index constant at about 1.06) and the spore volume probably increases exponentially. ...199810099291
morphological change and enhanced pigment production of monascus when cocultured with saccharomyces cerevisiae or aspergillus oryzaewhen a monascus isolate, a producer of monascus pigments, was cocultured with either saccharomyces cerevisiae or aspergillus oryzae in a solid sucrose medium, there were significant morphological changes in monascus culture. cocultures exhibited cell mass increases of 2 times and pigment yield increases of 30 to 40 times compared to monocultures of monascus. however, enhanced cell growth, an increase in pigment production, and morphological change did not occur in coculture with bacillus cereus. ...199810099374
enzymatic transformations in supersaturated substrate solutions: i. a general study with glycosidases.the results of an initial study of enzymatic catalysis in metastable supersaturated solutions of carbohydrates are presented. it has been shown that such solutions, formed in the presence of small amounts of water and alcohol as plasticizers, are sufficiently stable under ambient conditions to enable enzymatic transformations of substrates. a partial phase diagram for a system consisting of glucose, water, and (poly)ethylene glycol was constructed to identify the regions which are most suitable ...199810099420
metabolic engineering: techniques for analysis of targets for genetic manipulations.metabolic engineering has been defined as the purposeful modification of intermediary metabolism using recombinant dna techniques. with this definition metabolic engineering includes: (1) inserting new pathways in microorganisms with the aim of producing novel metabolites, e.g., production of polyketides by streptomyces; (2) production of heterologous peptides, e.g., production of human insulin, erythropoitin, and tpa; and (3) improvement of both new and existing processes, e.g., production of a ...199810191381
13c tracer experiments and metabolite balancing for metabolic flux analysis: comparing two approachesconventional metabolic flux analysis uses the information gained from determination of measurable fluxes and a steady-state assumption for intracellular metabolites to calculate the metabolic fluxes in a given metabolic network. the determination of intracellular fluxes depends heavily on the correctness of the assumed stoichiometry including the presence of all reactions with a noticeable impact on the model metabolite balances. determination of fluxes in complex metabolic networks often requir ...199810191397
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 ...199816961201
evolutionary relationships among aspergillus oryzae and related species based on the sequences of 18s rrna genes and internal transcribed spacers. 199812501432
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 ...199827392587
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.199827392399
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 ...199910196744
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 ...199910212209
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 ...199910215597
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 ...199910215899
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 ...199910227147
[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 ...199910228384
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 ...199910356263
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.199910359899
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 ...199910361683
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 ...199910375131
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 ...199910380624
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 ...199910386619
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 ...199910407143
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.199910417583
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 ...199910427010
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 ...199910434188
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 ...199910440673
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 ...199910446194
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 ...199910446375
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.199910448329
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 ...199910471703
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 ...199910473389
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 ...199910501004
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.199910540753
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 ...199910543807
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 ...199910099554
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 ...199910099591
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 ...199910099618
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 ...199910101277
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 ...199910101286
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: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 ...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
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 ...199922062704
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 ...199910543813
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