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a quick solution: ab initio structure determination of a 19 kda metalloproteinase using acorn.a data set from the metalloproteinase deuterolysin was collected at atomic resolution (1.0 a) with synchrotron radiation. the high resolution allowed the structure to be solved with the new direct-methods program acorn using the coordinates of the zn atom as a starting point. the phases obtained from acorn were of sufficient quality to allow automated building to be carried out in arp/warp. minimal manual rebuilding of the model was required and the structure determination was completed using th ...200111679721
establishment of a hyper-protein production system in submerged aspergillus oryzae culture under tyrosinase-encoding gene (melo) promoter control.uv-mediated mutagenesis generated a high glucoamylase-producing mutant of aspergillus oryzae exhibiting strong melanization in solid-state culture. expression of the glucoamylase-encoding gene (glab), which is specifically expressed in solid-state culture, and the tyrosinase-encoding gene (melo), was analyzed using an e. coli beta-glucuronidase (gus) reporter assay to investigate this phenomenon. although no common regulation was found for melo and glab expression, the former was greatly enhance ...200111693910
a novel thermostable branching enzyme from an extremely thermophilic bacterial species, rhodothermus obamensis.a branching enzyme (ec 2.4.1.18) gene was isolated from an extremely thermophilic bacterium, rhodothermus obamensis. the predicted protein encodes a polypeptide of 621 amino acids with a predicted molecular mass of 72 kda. the deduced amino acid sequence shares 42-50% similarity to known bacterial branching enzyme sequences. similar to the bacillus branching enzymes, the predicted protein has a shorter n-terminal amino acid extension than that of the escherichia coli branching enzyme. the deduce ...200111778874
a new flavone glycoside, 5-hydroxy 7,3',4',5'-tetra-methoxyflavone 5-o-beta-d-xylopyranosyl-(1-->2)-alpha-l-rhamnopyranoside from bauhinia variegata linn.a new flavone glycoside m.f. c(30)h(36)o(15) m.p. 252-253 degrees c, [m]+ 636 (eims) was isolated from the acetone soluble fraction of the concentrated 95% ethanolic extract of the seeds of bauhinia variegata (linn). it was identified as 5-hydroxy7,3',4',5'-tetra-methoxyflavone 5-o-beta-d-xylopyranosyl-(1-->2)-alpha-l-rhamnopyranoside (1) by various colour reactions, chemical degradations and spectral techniques.200111783588
electrophoretic mobility shift scanning using an automated infrared dna sequencer.electrophoretic mobility shift assay (emsa) is widely used in the study of sequence-specific dna-binding proteins, including transcription factors and mismatch binding proteins. we have established a non-radioisotope-based protocol for emsa that features an automated dna sequencer with an infrared fluorescent dye (irdye) detection unit. our modification of the elec- trophoresis unit, which includes cooling the gel plates with a reduced well-to-read length, has made it possible to detect shifted ...200111730013
identification of catalytic and substrate-binding site residues in bacillus cereus atcc7064 oligo-1,6-glucosidase.three active site residues (asp199, glu255, asp329) and two substrate-binding site residues (his103, his328) of oligo-1,6-glucosidase (ec 3.2.1.10) from bacillus cereus atcc7064 were identified by site-directed mutagenesis. these residues were deduced from the x-ray crystallographic analysis and the comparison of the primary structure of the oligo-1,6-glucosidase with those of saccharomyces carlsbergensis alpha-glucosidase, aspergillus oryzae alpha-amylase and pig pancreatic alpha-amylase which ...200111676021
isoflavone transformation during soybean koji preparation and subsequent miso fermentation supplemented with ethanol and nacl.soybeans were soaked with water for 4 h, steam-cooked, inoculated with the conidia of aspergillus oryzae, and incubated for 3 days for koji preparation. the koji was then mixed with water-soaked and steam-cooked soybeans (1:2, w/w), ground into paste, and supplemented with 15% ethanol and 12.5% nacl or 3% ethanol and 6% nacl for miso fermentation at 30 degrees c. daidzin, genistin, daidzein, and genistein contents were extracted from the lyophilized and pulverized soybean powder or from the miso ...200111513643
visualization of nuclei in aspergillus oryzae with egfp and analysis of the number of nuclei in each conidium by facs.aspergillus oryzae has been reported to form conidia with multinuclei. in order to analyze nuclei in living cells, we developed an expression system of the a. nidutans histone h2b protein tagged by egfp (h2b::egfp). in both a. oryzae niad300 and a. nidulans fgsc89 transformants expressing h2b::egfp, fluorescence was detected in nuclear regions of hyphae and conidia. while a conidium contained only one fluorescent spot in the a. nidulans transformant, approximately 66% of conidia had two, 24% had ...200111515532
a new class of glutaminase from aspergillus oryzae.the koji mold aspergillus oryzae is able to produce glutaminase which converts glutamine to glutamic acid, one of the most important flavor components in soy sauce. we present here the isolation and the complete nucleotide sequence of the glutaminase- encoding gene from a. oryzae u212, an industrial strain used in thailand. n-terminal and internal amino acid sequences were determined from purified glutaminase. a 700-bp fragment was amplified by pcr using oligonucleotide primers designed from par ...200111545278
increased phospholipid transfer protein activity in aspergillus oryzae grown on various industrial phospholipid sources.the effect of industrial carbon sources on phospholipid transfer protein production was investigated. phospholipid fractions of different composition were prepared from various plant oils (i.e., soybean, rapeseed, and sunflower) according to the lucas meyer extraction and purification process. the effect of these fractions on phospholipid transfer protein activity of cell extracts from aspergillus oryzae grown on medium containing these phospholipids as sole carbon source was studied. it was sho ...200111547891
[mystery of fermented foods--there are friendly molds too!]. 200111441805
on the involvement of electron transfer reactions in the fluorescence decay kinetics heterogeneity of proteins.time-resolved fluorescence study of single tryptophan-containing proteins, nuclease, ribonuclease t1, protein g, glucagon, and mastoparan, has been carried out. three different methods were used for the analysis of fluorescence decays: the iterative reconvolution method, as reviewed and developed in our laboratory, the maximum entropy method, and the recent method that we called "energy transfer" method. all the proteins show heterogeneous fluorescence kinetics (multiexponential decay). the orig ...200111567101
expression, gene cloning, and characterization of five novel phytases from four basidiomycete fungi: peniophora lycii, agrocybe pediades, a ceriporia sp., and trametes pubescens.phytases catalyze the hydrolysis of phosphomonoester bonds of phytate (myo-inositol hexakisphosphate), thereby creating lower forms of myo-inositol phosphates and inorganic phosphate. in this study, cdna expression libraries were constructed from four basidiomycete fungi (peniophora lycii, agrocybe pediades, a ceriporia sp., and trametes pubescens) and screened for phytase activity in yeast. one full-length phytase-encoding cdna was isolated from each library, except for the ceriporia sp. librar ...200111571175
plasminogen-binding activity of enolase in the opportunistic pathogen pneumocystis carinii.the glycolytic enzyme enolase is one of the most abundant proteins expressed in fungi and has been shown to be an immunodominant cell-wall-associated antigen of the pathogenic fungus, candida albicans. enolase has also been found on the surface of some mammalian cells where it functions as a plasminogen-binding motif and facilitator of plasminogen activation to plasmin. to investigate the immunogenicity of enolase in the opportunistic pathogen, pneumocystis carinii, the genomic and complementary ...200111798055
deletion analysis of the enolase gene (enoa) promoter from the filamentous fungus aspegillus oryzae.the enolase gene (enoa) is one of the most strongly expressed genes in aspergillus oryzae. to elucidate the transcription regulatory element for this strong expression and the process of glucose induction, the transcription activity of a series of truncated enoa promoters was measured by using the escherichia coli uida gene as a reporter. deletion of a 104-bp region located -224 nt to -121 nt upstream of the translation initiation site caused both a drastic decrease in the beta-glucuronidase (gu ...200111795846
a new biologically active triterpenoid saponin from the leaves of lepidagathis hyalina nees.a new biologically active triterpenoid saponin m.f. c42h68o13, m.p. 315 degrees-317 degrees c was isolated from the ethyl acetate soluble fraction of the methanolic extract of the leaves of lepidagathis hyalina. its structure was characterized as 3-beta-o-[alpha-l-rhamnopyranosyl(1-->4)o-beta-d-glucopyranosyl]16-alpha-hydroxy-olean-12-en(13)-28-oic acid by several spectral and chemical analysis. this new triterpenoid saponin showed antimicrobial activity against various plants pathogenic bacteri ...200111841115
the aspergillus niger faeb gene encodes a second feruloyl esterase involved in pectin and xylan degradation and is specifically induced in the presence of aromatic compounds.the faeb gene encoding a second feruloyl esterase from aspergillus niger has been cloned and characterized. it consists of an open reading frame of 1644 bp containing one intron. the gene encodes a protein of 521 amino acids that has sequence similarity to that of an aspergillus oryzae tannase. however, the encoded enzyme, feruloyl esterase b (faeb), does not have tannase activity. comparison of the physical characteristics and substrate specificity of faeb with those of a cinnamoyl esterase fro ...200211931668
immobilization of beta-galactosidase on fibrous matrix by polyethyleneimine for production of galacto-oligosaccharides from lactose.the production of galacto-oligosaccharides (gos) from lactose by aspergillus oryzae beta-galactosidase immobilized on cotton cloth was studied. a novel method of enzyme immobilization involving pei-enzyme aggregate formation and growth of aggregates on individual fibrils of cotton cloth leading to multilayer immobilization of the enzyme was developed. a large amount of enzyme was immobilized (250 mg/g support) with about 90-95% efficiency. a maximum gos production of 25-26% (w/w) was achieved at ...200211934291
molecular cloning, characterization, and expression analysis of the xynf3 gene from aspergillus oryzae.the gene encoding xylanase f3 (xynf3) was isolated from a genomic library of aspergillus oryzae kbn616, used for making shoyu koji. the structural part of xynf3 was found to be 1468 bp. the nucleotide sequence of cdna amplified by rt-pcr showed that the open reading frame of xynf3 was interrupted by ten short introns and encoded 323 amino acids. direct n-terminal amino acid sequencing showed that the precursor of xynf3 had a signal peptide of 22 amino acids. the predicted amino acid sequence of ...200211999400
the potential for establishment of axial temperature profiles during solid-state fermentation in rotating drum bioreactors.the mixing and heat transfer phenomena within rotating drum bioreactors (rdbs) used for solid-state fermentation processes are poorly studied. the potential for the establishment of axial temperature gradients within the substrate bed was explored using a heat transfer model. for growth of aspergillus oryzae on wheat bran within a 24 l rdb with air at a superficial velocity of 0.0023 m s(-1) and 15% relative humidity, the model predicts an axial gradient between the air inlet and outlet of 2 deg ...200212209792
cloning and structural analysis of bglm gene coding for the fungal cell wall-lytic beta-1,3-glucan-hydrolase bglm of bacillus circulans iam1165.bacillus circulans iam1165 produces isoforms of beta-1,3-glucan-hydrolases. of these enzymes, the 42-kda enzyme bgim degrades aspergillus oryzae cell walls the most actively. a gene coding for a bgim precursor consisting of 411 amino acid residues was cloned. the 27 n-terminal amino acid sequence of the precursor is a signal peptide. the 141 c-terminal amino acid sequence showed a motif of carbohydrate-binding module family 13. this domain bound to pachyman, lichenan, and a. oryzae cell walls. t ...200212162545
chsz, a gene for a novel class of chitin synthase from aspergillus oryzae.we cloned and characterized a novel aspergillus oryzae chitin synthase gene, chsz, encoding a polypeptide containing a new myosin motor-like domain in its n-terminal half. alignment analysis revealed that chsz was less homologous to known class v enzymes, except for its probable chitin synthase conserved region in the c-terminal half. we also found a chsy gene and found that chsy showed higher similarity to the class v enzymes than did chsz. phylogenetic analysis clearly demonstrated that the a. ...200212172967
water and glucose gradients in the substrate measured with nmr imaging during solid-state fermentation with aspergillus oryzae.gradients inside substrate particles cannot be prevented in solid-state fermentation. these gradients can have a strong effect on the physiology of the microorganisms but have hitherto received little attention in experimental studies. we report gradients in moisture and glucose content during cultivation of aspergillus oryzae on membrane-covered wheat-dough slices that were calculated from (1)h-nmr images. we found that moisture gradients in the solid substrate remain small when evaporation is ...200212209813
comparison of lysis methods and preparation protocols for one- and two-dimensional electrophoresis of aspergillus oryzae intracellular proteins.filamentous fungal fermentations are used to produce billions of dollars of biochemical and pharmaceutical products annually, yet are plagued by a number of poorly understood problems that would benefit from proteomic analysis. unfortunately, few publications are available which describe extraction of filamentous fungal proteins for two-dimensional electrophoresis. the goal here was to develop protocols for extraction of fungal proteins, from both wild-type and a recombinant strain of the indust ...200212210225
effect of molecular confinement on internal enzyme dynamics: frequency domain fluorometry and molecular dynamics simulation studies.the tryptophanyl emission decay of the mesophilic beta-galactosidase from aspergillus oryzae free in buffer and entrapped in agarose gel is investigated as a function of temperature and compared to that of the hyperthermophilic enzyme from sulfolobus solfataricus. both enzymes are tetrameric proteins with a large number of tryptophanyl residues, so the fluorescence emission can provide information on the conformational dynamics of the overall protein structure rather than that of the local envir ...200212073936
evaluation of filamentous fungi and inducers for the production of endo-polygalacturonase by solid state fermentation.aspergillus oryzae cct 3940, aspergillus awamori nrrl 3112 and a trichoderma sp.) were compared for their capacity to produce endo-polygalacturonase (endo-pg) in solid state fermentation. maximum pectinolytic activity was reached in 72 h of growth, the best two fungal strains being a. niger t0005007-2 and a. oryzae cct 3940. three types of commercial purified pectin and four of unprocessed pectin (tangerine, orange, tahiti lime and sweet lime rind) were used to assess the effect of pectin on the ...200212240994
biosynthesis of l-dopa by aspergillus oryzae.the present investigation deals with the biosynthesis of l-dopa by parental (gcb-6) and mutant (uv-7) strains of aspergillus oryzae. there was a marked difference between the mycelial morphology and pellet type of parental and uv-irradiated mutant culture. the mutant strain of a. oryzae uv-6 exhibited pellet-like mycelial morphology and improved tyrosinase activity. mould mycelium was used for biochemical conversion of l-tyrosine to l-dopa because tyrosinase is an intracellular enzyme. the mutan ...200212146638
estimation of hyphal tensile strength in production-scale aspergillus oryzae fungal fermentations.fragmentation of filamentous fungal hyphae depends on two phenomena: hydrodynamic stresses, which lead to hyphal breakage, and hyphal tensile strength, which resists breakage. the goal of this study was to use turbulent hydrodynamic theory to develop a correlation that allows experimental data of morphology and hydrodynamics to be used to estimate relative (pseudo) tensile strength (sigma(pseudo)) of filamentous fungi. fed-batch fermentations were conducted with a recombinant strain of aspergill ...200211807755
a transcriptional activator, aoxlnr, controls the expression of genes encoding xylanolytic enzymes in aspergillus oryzae.by deletion across the promoter region of the xynf1 gene encoding the major aspergillus oryzae xylanase, a 53-bp dna fragment containing the xlnr binding sequence ggctaaa as well as two similar sequences was shown to confer xylan inducibility on the gene. complementary and genomic dnas encoding the aspergillus niger xlnr homologous gene, abbreviated aoxlnr, were cloned from a. oryzae and sequenced. aoxlnr comprised 971 amino acids with a zinc binuclear cluster domain at the n-terminal region and ...200211848678
immobilization of an l-aminoacylase-producing strain of aspergillus oryzae into gelatin pellets and its application in the resolution of d,l-methionine.the conditions for immobilization of an l-aminoacylase-producing strain of aspergillus oryzae in gelatin and the enzymic characteristics of the immobilized pellets were studied. the optimal concentrations of gelatin, glutaraldehyde and ethyldiamine and time of immobilization were determined. scanning electron micrographs reveal the cross-linked structure differences between the native and immobilized pellets. optimum ph and temperature of the native and immobilized pellets were determined. effec ...200211916452
dependence of morphology on agitation intensity in fed-batch cultures of aspergillus oryzae and its implications for recombinant protein production.we previously reported that, although agitation conditions strongly affected mycelial morphology, such changes did not lead to different levels of recombinant protein production in chemostat cultures of aspergillus oryzae (amanullah et al., 1999). to extend this finding to another set of operating conditions, fed-batch fermentations of a. oryzae were conducted at biomass concentrations up to 34 g dry cell weight/l and three agitation speeds (525, 675, and 825 rpm) to give specific power inputs b ...200211835142
production of galacto-oligosaccharides from lactose by aspergillus oryzae beta-galactosidase immobilized on cotton cloth.the production of galacto-oligosaccharides (gos) from lactose by a. oryzae beta-galactosidase immobilized on cotton cloth was studied. the total amounts and types of gos produced were mainly affected by the initial lactose concentration in the reaction media. in general, more and larger gos can be produced with higher initial lactose concentrations. a maximum gos production of 27% (w/w) of initial lactose was achieved at 50% lactose conversion with 500 g/l of initial lactose concentration. tri-s ...200211745169
behaviors of d- and l-lactic acids during the brewing process of sake (japanese rice wine).the amounts of d- and l-lactic acids during the brewing process of sake were determined by capillary electrophoresis using 2-hydroxypropyl-beta-cyclodextrin as a chiral selector. because l-lactic acid, which prevents the growth of nonuseful microorganisms, is a raw material of sake, the ratio of l-lactic acid to total lactic acid is almost 1.0 at the initial stage of sake brewing. during brewing, the ratio decreased gradually and finally reached 0.39. yeast (saccharomyces cerevisiae) for sake br ...200211829643
expression of an alpha-galactosidase from saccharomyces cerevisiae in aspergillus awamori and aspergillus oryzae.a gene encoding alpha-galactosidase activity was isolated by polymerase chain reaction (pcr) from saccharomyces cerevisiae ncyc686 and separately placed under the control of transcriptional elements regulating alpha-amylase expression in aspergillus oryzae and glucoamylase expression in a. awamori. following transformation of both a. oryzae and a. awamori with their respective expression vectors, induction of heterologous alpha-galactosidase from positively selected clones was effected through t ...200212074058
purification and characterization of an extracellular protease from penicillium chrysogenum pg222 active against meat proteins.an extracellular protease from penicillium chrysogenum (pg222) isolated from dry-cured ham has been purified. the purification procedure involved several steps: ammonium sulfate precipitation, ion-exchange chromatography, filtration, and separation by high-performance liquid chromatography. based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (sds-page) analysis and gel filtration, the purified fraction showed a molecular mass of about 35 kda. the hydrolytic properties of the purif ...200212089038
molecular cloning and overexpression of flea gene encoding a fucose-specific lectin of aspergillus oryzae.a protein from the cell lysate of aspergillus oryzae was purified by column chromatography immobilized with a ferrichrysin (fcy), which is one of the siderophores of a. oryzae. it is produced only in an iron-deficient culture and its molecular weight is estimated as 35,000 by sds-page. two internal amino acid sequences of the protein obtained by lysylendopeptidase digestion were analyzed. molecular cloning shows that it encodes 310 putative amino acid residues separated by 4 introns and is desig ...200212092808
[study of immobilization of aspergillus oryzae mycelium aided by artificial neural network].aspergillus oryzae mycelium pellets with abundant aminoacylase were cultured by liquid fermentation. mycelium pellets were immobilized by crosslinking method with reagent of gelatin and formaldehyde. basing on the orthogonal design table l16 (4(5)), artificial neural network(ann) model with back-propagation(bp) of error structured 4-10-15-1 was used to optimize the immobilization conditions. and the optimal conditions were obtained. then the activity of the immobilized cells prepared under the o ...200212148288
functional expression in aspergillus oryzae of p15, a protein with potent neurite-inducing activity in pc12 cells.we previously reported that a fungal protein, p15, induces neurite outgrowth and differentiation of rat pheochromocytoma pc12 cells through the activation of the ca2+ signaling pathway. we report here the secretory production of p15 in aspergillus oryzae. analysis of culture supernatant of a. oryzae transformed with the gene encoding the p15 precursor tagged with a hemagglutinin (ha) epitope demonstrated that the transformant secreted a protein with an apparent molecular mass of 17.5 kda, which ...200212005070
a simple method for enrichment of uninucleate conidia of aspergillus oryzae.aspergillus oryzae produces multinucleate conidia, which makes the obtaining of homokaryons labor-intensive. analysis of conidia by flow cytometry clarified the relationship that conidia of lower nuclear number were smaller in size. based on this, we have developed a simple way to enrich uninucleate conidia with a membrane filter. our results also suggest that the method is useful for elimination of heterokaryons.200212005075
microbiological transformation of l-tyrosine to 3,4-dihydroxyphenyl l-alanine (l-dopa) by a mutant strain of aspergillus oryzae uv-7.the present study deals with the microbiological transformation of l-tyrosine to 3,4-dihydroxyphenyl l-alanine by a mutant strain of aspergillus oryzae uv-7. sixteen different mutant strains of aspergillus oryzae (gcb-6) were isolated through uv-irradiation. these mutant strains were screened for the production of mold mycelia by submerged fermentation in 250-ml erlenmeyer flasks. of all the mutant strains examined, uv-7 gave maximum production of l-dopa (1.28 mg/ml). the reaction was carried ou ...200212070684
cloning and characterization of vmaa, the gene encoding a 69-kda catalytic subunit of the vacuolar h+-atpase during alkaline ph mediated growth of aspergillus oryzae.screening of a cdna library constructed under alkaline ph mediated growth of aspergillus oryzae implicated a vacuolar h+-atpase gene (vmaa) as a putative candidate involved in alkaline ph adaptation. a. oryzae vmaa genomic dna extended to 2072 bp including three introns and encoded a protein of 605 amino acids. vmaap was homologous to vma-1p from neurospora crassa (71%), vma1p from saccharomyces cerevisiae (69%) and atp6a2 from human (49%). the vmaa cdna complemented s. cerevisiae v-atpase disru ...200212007818
polymerase chain reaction-mediated characterization of molds belonging to the aspergillus flavus group and detection of aspergillus parasiticus in peanut kernels by a multiplex polymerase chain reaction.the aspergillus flavus group covers species of a. flavus and aspergillus parasiticus as aflatoxin producers and aspergillus oryzae and aspergillus sojae as koji molds. genetic similarity among these species is high, and aflatoxin production of a culture may be affected by cultivation conditions and substrate composition. therefore, a polymerase chain reaction (pcr)-mediated method of detecting the aflatoxin-synthesizing genes to indicate the degree of risk a genotype has of being a phenotypic pr ...200212030297
cloning and expression of the exo-beta-d-1,3-glucanase gene (exgs) from aspergillus saitoi.a gene of exo-1,3-beta-d-glucanase (exgs) was cloned from a koji mold, aspergillus saitoi, genomic dna using pcr. the exgs has an orf comprising 2832 bp, which contains one intron of 45 bp, and encodes 945 amino acids. the deduced amino acid sequences showed that the exgs has a non-homologous linker region consisting of 180 amino acids, which encompassed highly conserved regions observed in exg homologues from filamentous fungi. a recombinant protein (exgs) has been recovered from the cultural f ...200212224649
subtractive cloning of cdna from aspergillus oryzae differentially regulated between solid-state culture and liquid (submerged) culture.in solid-state cultures (sc), aspergillus oryzae shows characteristics such as high-level production and secretion of enzymes and hyphal differentiation with asexual development which are absent in liquid (submerged) culture (lc). it was predicted that many of the genes involved in the characteristics of a. oryzae in sc are differentially expressed between sc and lc. we generated two subtracted cdna libraries with bi-directional cdna subtractive hybridizations to isolate and identify such genes. ...200212172969
toxicological studies on polyporus pinsitus laccase expressed by aspergillus oryzae intended for use in food.the laccase used in the study was produced by submerged fermentation of aspergillus oryzae, containing a gene originating from polyporus pinsitus. laccase is to be employed as a processing aid in the juice industry to make a clear and stable juice. the enzyme was subject to a series of toxicological tests to document its safety in use. it was not mutagenic in the salmonella typhimurium reverse mutation assay, and it did not cause chromosomal aberrations in cultured human lymphocytes. no evidence ...200211962689
a polymerase chain reaction-based method for cloning novel members of a gene family using a combination of degenerate and inhibitory primers.we have developed a novel method for cloning gene family members by using a polymerase chain reaction technique. the method is based on the amplification of a broad range of homologous genes in combination with the specific inhibition of already cloned genes. to accomplish this, we designed degenerate primers to highly conserved regions among the gene family members, and inhibitory primers to the divergent region at the 3'-margin of each degenerate primer. the 5'-end of the inhibitory primer, th ...200212036596
effects of increased impeller power in a production-scale aspergillus oryzae fermentation.the goal in this study was to determine how increased impeller power affects enzyme expression in large-scale (80 m(3)), fed-batch aspergillus oryzae fermentations. an approximate 50% increase in average impeller power was achieved by increasing impeller diameter approximately 10%, while operating at slightly reduced speed. measured decreases in terminal (95%) mixing time show increased power improved bulk mixing. however, batches operated at increased power had lower recombinant enzyme producti ...200212052056
contribution of aerial hyphae of aspergillus oryzae to respiration in a model solid-state fermentation system.oxygen transfer is for two reasons a major concern in scale-up and process control in industrial application of aerobic fungal solid-state fermentation (ssf): 1) heat production is proportional to oxygen uptake and it is well known that heat removal is one of the main problems in scaled-up fermenters, and 2) oxygen supply to the mycelium on the surface of or inside the substrate particles may be hampered by diffusion limitation. this article gives the first experimental evidence that aerial hyph ...200212115123
synthesis and utility of sulfated chromogenic carbohydrate model substrates for measuring activities of mucin-desulfating enzymes.a chromogenic substrate, 4-nitrophenyl 2-acetamido-2-deoxy-beta-d-glucopyranoside 6-sodium sulfate was synthesized and used in combination with beta-n-acetylhexosaminidase for detection of the sulfatase, mdsa, by release of 4-nitrophenol. mdsa was originally isolated from the bacterium prevotella strain rs2 and is believed to be involved in desulfation of sulfomucins, major components of the mucus barrier protecting the human colon surface. the exo nature of the mdsa sulfatase was indicated by i ...200212062525
antifungal efficacy of bacteria isolated from marine sedentary organisms.the antibiotic-producing ability of 57 bacteria isolated from 8 marine sedentary organisms, 6 sponges (spirastrella sp., phyllospongia sp., ircinia sp., aaptos sp., azorica sp., axinella sp.), 1 soft coral (lobophytum sp.) and 1 alga (sargassum sp.), was evaluated against 6 phytopathogenic fungi (helminthosporium oryzae, rhizoctonium solani, pyricularia oryzae, fusarium oxysporum, aspergillus oryzae and a. fumigatus). bacteria of the genus bacillus (20%), pseudomonas (33%) and flavobacterium (40 ...200211980270
nonfunctionality of aspergillus sojae aflr in a strain of aspergillus parasiticus with a disrupted aflr gene.aspergillus sojae belongs to the aspergillus section flavi but does not produce aflatoxins. the functionality of the a. sojae aflr gene (aflrs) was examined by transforming it into an deltaaflr strain of a. parasiticus, derived from a nitrate-nonutilizing, versicolorin a (vera)-accumulating strain. the a. parasiticus aflr gene (aflrp) transformants produced vera, but the aflrs transformants did not. even when aflrs was placed under the control of the amylase gene (amyb) promoter of aspergillus o ...200212147467
visualization of vacuoles in aspergillus oryzae by expression of cpy-egfp.vacuolar carboxypeptidase y (cpy) from aspergillus nidulans was used to construct a cpy-egfp fusion protein and expressed in a. oryzae to study vacuolar morphology and functions in a. oryzae. while the fluorescence of egfp was barely detectable in a. oryzae expressing cpy-egfp grown under normal conditions at ph 5-6, the increase in ph of the growth medium towards alkalinity restored the fluorescence. in accordance with such an observation, the fluorescence of cpy-egfp fusion protein in cell ext ...200212223187
toxicological studies on laccase from myceliophthora thermophila expressed in aspergillus oryzae.the bioindustrially produced enzyme laccase can be used in different technical and food applications to facilitate processes. it can be added to different oral care products such as mouthwash, toothpaste, mints, and gums to prevent halitosis. laccase, produced by submerged fermentation of aspergillus oryzae, containing a gene originating from myceliophthora thermophila, was subject to a series of toxicological tests to document its safety in use. it was not found to be mutagenic in the salmonell ...200212202045
transformation of aspergillus sp. and trichoderma reesei using the pyrithiamine resistance gene (ptra) of aspergillus oryzae.a pyrithiamine (pt) resistance gene (ptra) was cloned from a pt resistant mutant of aspergillus oryzae and was useful as a dominant selectable marker for transformation of all a. oryzae wild type strain as well as a. nidulans. for further study, we examined whether or not ptra could be used as the transformation marker in other species of filamentous fungi. two types of plasmid, which contain ptra as a selectable marker, were constructed, and the transformation experiments were done with them. o ...200211999416
preventive effect of fermented brown rice and rice bran against colon carcinogenesis in male f344 rats.epidemiological and preclinical studies demonstrate that nutrition plays an important role in the etiology of cancer. it has been reported that rice components, especially rice germ plays a key role in prevention of cancer. the experiments described here examined the potential anticancer properties of brown rice fermented by aspergillus oryzae (fbra) in male f344 rats using inhibition of the formation of azoxymethene (aom) induced aberrant crypt foci (acf) and tumors in the colon as the measure ...200212066215
reactivity and stability of mycelium-bound carboxylesterase from aspergillus oryzae.the reactivity and thermostability of a novel mycelium-bound carboxylesterase from lyophilized cells of aspergillus oryzae are explored in organic solvent. ethanol acetylation was selected as reference esterification reaction. high carboxylesterase activity cells were used as biocatalyst in batch esterification tests at 12.5 < s(o) < 125 mmol l(-1), 5.0 < x(o) < 30 g l(-1), 0.49 < log p < 4.5 and 30 < t < 80 degrees c, as well as in residual activity tests after incubation at 40 < t < 90 degrees ...200211753931
impact of four (13)c-proline isotope labels on the infrared spectra of ribonuclease t1.ribonuclease t1 was biosynthesized, with all four prolines (13)c-labeled in the peptide c[double bond]o bond, using a proline auxotrophic yeast strain of saccharomyces cerevisiae. the (13)c- and (12)c-proline isotopomers of ribonuclease t1 were investigated by infrared spectroscopy in the thermally unfolded and natively folded state at 80 and 20 degrees c, respectively. in the thermally unfolded state, both proteins established almost indistinguishable spectral features in the secondary structur ...200212033852
validation of a model for process development and scale-up of packed-bed solid-state bioreactors.we have validated our previously described model for scale-up of packed-bed solid-state fermenters (weber et al., 1999) with experiments in an adiabatic 15-dm(3) packed-bed reactor, using the fungi coniothyrium minitans and aspergillus oryzae. effects of temperature on respiration, growth, and sporulation of the biocontrol fungus c. minitans on hemp impregnated with a liquid medium were determined in independent experiments, and the first two effects were translated into a kinetic model, which w ...200211787011
metabolic engineering of the morphology of aspergillus oryzae by altering chitin synthesis.morphology and alpha-amylase production during submerged cultivation were examined in a wild-type strain (a1560) and in strains of aspergillus oryzae in which chitin synthase b (chsb) and chitin synthesis myosin a (csma) have been disrupted (chsb/g and cm101). in a flowthrough cell, the growth of submerged hyphal elements was studied online, making it possible to examine the growth kinetics of the three strains. the average tip extension rates of the cm101 and chsb/g strains were 25 and 88% lowe ...200211916702
physiological characterisation of recombinant aspergillus nidulans strains with different crea genotypes expressing a. oryzae alpha-amylase.the physiology of three strains of aspergillus nidulans was examined--a crea deletion strain, a wild type crea genotype and a strain containing extra copies of the crea gene, all producing aspergillus oryzae alpha-amylase. the strains were cultured in batch and continuous cultivations and the biomass formation and alpha-amylase production was characterised. overexpression of the crea gene resulted in a lower maximum specific growth rate and a slightly higher repression of the alpha-amylase produ ...200211689252
generic model of morphological changes in growing colonies of fungi.fungal colonies are able to exhibit different morphologies depending on the environmental conditions. this allows them to cope with and adapt to external changes. when grown in solid or semisolid media the bulk of the colony is compact and several morphological transitions have been reported to occur as the external conditions are varied. here we show how a unified simple mathematical model, which includes the effect of the accumulation of toxic metabolites, can account for the morphological cha ...200211863559
observation of egfp-visualized nuclei and distribution of vacuoles in aspergillus oryzae arpa null mutant.the arpa gene encoding arp1 (actin-related protein) was previously cloned and characterized from aspergillus oryzae. phenotypes of the arpa null mutant indicate its requirement for normal nuclear distribution and morphology of conidiophores. in this study, we further characterized the function of the arpa gene in distribution of organelles. for further analysis of nuclear migration in living cells, an expression system consisting of a fusion protein of aspergillus nidulans histone h2b and egfp ( ...200211786257
production of cellulose- and xylan-degrading enzymes by a koji mold, aspergillus oryzae, and their contribution to the maceration of rice endosperm cell wall.the production of cellulose- (cel), xylan- (xyl), and pectin-degrading enzymes (pec) by a koji mold, aspergillus oryzae, was studied, and their contributions to the maceration of the rice endosperm cell wall were investigated with regard to the utilization of available rice in the sake mash. the sake koji mold showed higher cel and xyl productivities, whereas the miso and soy sauce koji molds showed higher pec productivity. statistical analyses indicated that cel and xyl contribute predominantly ...200216233157
enzymatic synthesis of l-menthyl alpha-maltoside and l-menthyl alpha-maltooligosides from l-menthyl alpha-glucoside by cyclodextrin glucanotransferase.l-menthyl alpha-d-glucopyranosyl-(1-->4)-alpha-d-glucopyranoside (alpha-meng2), a novel glycoside of l-menthol, was synthesized enzymatically and its physicochemical properties were characterized. production of alpha-meng2 from l-menthyl alpha-d-glucopyranoside (alpha-meng) was attempted since we had already succeeded in the high-yield production of alpha-meng using a xanthomonas campestris enzyme (nakagawa h., et al. j. biosci. bioeng., 89, 138-144, 2000). through production tests on enzymes, i ...200216233280
recent studies of protein secretion by filamentous fungi.filamentous fungi have been widely exploited for the homologous and heterologous protein production, because of the high capacity of their protein secretion machinery. however, the production of heterologous proteins is often limited while the production of homologous proteins can be very high. various researches have reported the methods for overcoming this problem and some techniques, such as the fusion gene system, improve the production of heterologous proteins. recently, the molecular biolo ...200216233346
simultaneous display of bacterial and fungal lipases on the cell surface of bacillus subtilis.a small cell wall-binding domain (cwb(c)) of the bacillus subtilis peptidoglycan hydrolase cwlc fused to b. subtilis lipase b was able to be localized on the cell wall of b. subtilis. with the aim of developing an efficient lipid-hydrolyzing bacterium with two different lipases on the cell surface, plasmids possessing genes for lipb-cwb(c) and cwb(b)-cutl (the cell wall-binding domain of cwlb fused to the aspergillus oryzae lipolytic enzyme cutl) were constructed. b. subtilis harboring these pla ...200216233158
properties of cellulose-degrading enzymes from aspergillus oryzae and their contribution to material utilization and alcohol yield in sake mash fermentation.four cellulose-degrading enzymes were identified in a solid-state culture of aspergillus oryzae. the three major enzymes were purified and named cel-1, cel-2, and cel-3, respectively. the molecular weights were determined to be 62, 120, and 34 kda, respectively. the optimum temperature of cel-3 activity was higher than that of the other enzymes. an acidic ph was found to be more suitable for cel-1 activity than for the other enzymes, and cel-3 was more stable under acidic conditions than the oth ...200216233235
specific expression and temperature-dependent expression of the acid protease-encoding gene (pepa) in aspergillus oryzae in solid-state culture (rice-koji).the synthesis of acid protease in rice-koji is important for sake brewing. northern blot analysis was carried out to study the transcriptional regulation of acid protease-encoding gene (pepa in aspergillus orytae. the pepa gene was not expressed in submerged culture, while it was expressed when cultured on steamed rice. additionally, the culture at high temperature (>38 degrees c) caused a marked decrease in transcription level of pepa, although the alpha-amylase (amyb) and actin genes were expr ...200216233250
intake, digestion, and digestive characteristics of neotyphodium coenophialum-infected and uninfected fescue by heifers offered hay diets supplemented with aspergillus oryzae fermentation extract or laidlomycin propionate.tarentaise heifers fitted with a rumen cannula (539 +/- 7.5 and 487 +/- 15.7 kg avg initial bw in exp. 1 and 2, respectively) were used in two latin square metabolism experiments having 2 x 2 factorial treatment arrangements to determine the effects of supplementation with aspergillus oryzae fermentation extract (ao) or laidlomycin propionate (lp) on intake, digestion, and digestive characteristics of neotyphodium coenophialum-infected (if) or uninfected (ff) tall fescue (festuca arundinacea) ha ...200211831521
statistical media optimization and production of its alpha-amylase from aspergillus oryzae in a bioreactor.the production of an intermediate temperature-stable (its) alpha-amylase from aspergillus oryzae was studied by using a central composite design with three independent variables, viz., starch, yeast extract, and k(2)hpo(4). the model equation provided a suitable model for the response surface for alpha-amylase production, and, from the optimal concentrations of the medium components, a model was predicted, which was then used for enzyme production in a 150-l bioreactor. in the bioreactor studies ...200212177743
cloning, nucleotide sequencing, and expression of the beta-galactosidase-encoding gene (laca) from aspergillus oryzae.laca coding for beta-galactosidase (beta-gal) was cloned from the genomic dna of aspergillus oryzae rib40. there were 9 exons in laca and the coding region of 3,015 bp encoded a protein of 1,005 aa with a deduced molecular mass of 109,898. a. oryzae laca was highly homologous to fungal beta-gals, with the highest aa identity of 70.7% to a. niger laca, and also showed significant identity to acid beta-gals belonging to family 35 glycosyl hydrolases. approximately 10 copies of laca under control o ...200212469296
secreted metalloprotease gene family of microsporum canis.keratinolytic proteases secreted by dermatophytes are likely to be virulence-related factors. microsporum canis, the main agent of dermatophytosis in dogs and cats, causes a zoonosis that is frequently reported. using aspergillus fumigatus metalloprotease genomic sequence (mep) as a probe, three genes (mep1, mep2, and mep3) were isolated from an m. canis genomic library. they presented a quite-high percentage of identity with both a. fumigatus mep and aspergillus oryzae neutral protease i genes. ...200212228297
evaluation of antioxidative and mutagenic properties of 50% ethanolic extract from red beans fermented by aspergillus oryzae.various bean products fermented by microorganisms are commonly consumed in asian diets; however, the safety or functional properties of fermented beans can vary with different microbial species and with different processes being applied to different beans. the objectives of this study were to evaluate the antioxidative and mutagenic properties of 50% ethanolic extracts from red beans fermented by aspergillus oryzae. the extracts' antioxidative activities, including alpha,alpha;-diphenyl-beta-pic ...200212233859
molecular biology of the koji molds. 200212236056
progress of aspergillus oryzae genomics. 200212236061
transcriptional activator, aoxlnr, mediates cellulose-inductive expression of the xylanolytic and cellulolytic genes in aspergillus oryzae.aoxlnr was isolated as a transcriptional activator of the major xylanase gene, xynf1, in aspergillus oryzae. to investigate the spectrum of genes under the control of aoxlnr, expression of the xylanolytic and cellulolytic genes in an a. oryzae wild type strain, an aoxlnr disruptant and an aoxlnr overexpressed strain was analyzed by northern blotting. aoxlnr mediated expression of at least four xylanolytic genes and four cellulolytic genes when induced by xylan and d-xylose. moreover, aoxlnr was ...200212297320
synthesis of alpha-amylase by aspergillus oryzae in solid-state fermentation.spent brewing grains (sbg) was evaluated for its efficacy to be used as sole carbon source for the synthesis of alpha-amylase in solid-state fermentation using a fungal strain of aspergillus oryzae nrrl 6270. enzyme production was superior when the culture grew on mesophilic temperatures and best yields were at 25 degrees c. at 30 degrees c, yields were almost comparable. maximum production of alpha-amylase [6870 u/g dry substrate (gds)] was obtained when ssf was carried out at 30 degrees c for ...200212362403
effect of ph on simultaneous saccharification and isomerization by glucoamylase and glucose isomerase.ph and temperature play critical roles in multistep enzymatic conversions. in such conversions, the optimal ph for individual steps differs greatly. in this article, we describe the production of glucoamylase (from aspergillus oryzae mtcc152 in solid-state fermentation) and glucose isomerase (from streptomyces griseus ncim2020 in submerged fermentation), used in industries for producing high-fructose syrup. optimum ph for glucoamylase was found to be 5.0. for glucose isomerase, the optimum ph ra ...200212396122
production of alpha-amylase with aspergillus oryzae on spent brewing grain by solid substrate fermentation.ten aspergillus oryzae strains were screened in solid substrate fermentation for alpha-amylase production on spent brewing grain (sbg) and on corn fiber. sbg proved to be a better substrate for enzyme production than corn fiber. a plackett-burman experimental design was used to optimize the medium composition for the best strain. solid substrate fermentation on optimized medium with a. oryzae nrrl 1808 (=atcc 12892) strain in stationary 500-ml erlenmeyer flask culture yielded 4519 u of alpha-amy ...200212396145
determination of platinated purines in oligoribonucleotides by limited digestion with ribonucleases t1 and u2.platinum complexes which are known to react preferentially with guanine (g) and adenine (a) bases of oligonucleotides can be used as tools to analyze their tertiary structures and eventually to cross-link them. however, this requires efficient methods to allow the identification and quantification of the corresponding adducts which have so far been developed only for oligodeoxyribonucleotides. maxam-gilbert type digestions cannot be used for rnas and hplc techniques would require too large amoun ...200212413471
a protease-activated pathway underlying th cell type 2 activation and allergic lung disease.the respiratory allergens that induce experimental th cell type 2-dependent allergic lung inflammation may be grouped into two functional classes. one class of allergens, in this study termed type i, requires priming with adjuvants remote from the lung to overcome airway tolerogenic mechanisms that ordinarily preclude allergic responses to inhaled ags. in contrast, the other, or type ii, allergen class requires neither remote priming nor additional adjuvants to overcome airway tolerance and elic ...200212421974
differentiation of feruloyl esterases on synthetic substrates in alpha-arabinofuranosidase-coupled and ultraviolet-spectrophotometric assays.4-nitrophenyl 5-o-trans-feruloyl-alpha-l-arabinofuranoside and 4-nitrophenyl 2-o-trans-feruloyl-alpha-l-arabinofuranoside, synthesized by our group (m. mastihubová, j. szemesová, and p. biely), were found to be suitable substrates for determination of activity of feruloyl esterases (fees) exhibiting affinity for 5-o- and 2-o-feruloylated alpha-l-arabinofuranosyl residues. one assay is based on coupling the fee-catalyzed formation of 4-nitrophenyl alpha-l-arabinofuranoside with its efficient hydr ...200212441154
cloning and characterization of the naga gene that encodes beta-n-acetylglucosaminidase from aspergillus nidulans and its expression in aspergillus oryzae.we isolated a beta-n-acetylglucosaminidase encoding gene and its cdna from the filamentous fungus aspergillus nidulans, and designated it naga. the naga gene contained no intron and encoded a polypeptide of 603 amino acids with a putative 19-amino acid signal sequence. the deduced amino acid sequence was very similar to the sequence of candida albicans hex1 and trichoderma harzianum nag1. yeast cells containing the naga cdna under the control of the gal1 promoter expressed beta-n-acetylglucosami ...200212450128
[improvement of recombinant xylanase xynf1 from aspergillus oryzae expressed in pichia pastoris by site-directed mutagenesis].the recombinant xylanase xynf1 from aspergillus oryzae expressed in the methylotrophic yeast pichia pastoris was improved by site-directed mutagenesis. the mutant i156a secreted about 47.7 u/ml xylanase xynf1 in the bmmy medium. the mutant xylanase xynf1 was purified by ion exchange and gel filtration chromatographies. the molecular weight of purified xynf1 was 35 kd. xynf1 was stable between ph 4-9 and its optimum ph was 7.0. the optimum temperature of xynf1 was 45 degrees c and it was stable b ...200212557547
[cloning and sequencing of lactase gene from aspergillus candidus].genomic dna and cdna sequences of lactase from aspergillus candidus were cloned. sequences analysis revealed that the genomic dna was 3458 bp containing eight introns, cdna was 3015 bp and encoding a polypeptide of 1005 amino acid residues. signal peptide was 19 amino acid residues, eleven potential n-glycosylation sites were assumed. comparing the gene cdna and amino acid sequences with other lactase sequences from various sources, it showed a very low homology with most of other sequences. alt ...200212561200
cloning and nucleotide sequence of the glutamate decarboxylase-encoding gene gada from aspergillus oryzae.we cloned a genomic dna encoding the glutamate decarboxylase (gad) from aspergillus oryzae using a 200-bp dna fragment as the probe. this dna fragment was amplified by the reverse transcription polymerase chain reaction with mrna of a. oryzae as the template and degenerate primers designed from the conserved amino acid sequence of escherichia coli gad and arabidopsis thaliana gad. nucleotide sequencing analysis showed that the cloned gene (designated gada) encoded 514 amino acid residues and con ...200212596854
antifungal activity of substituted 8-quinolinol-5- and 7-sulfonic acids: a mechanism of action is suggested based on intramolecular synergism.8-quinolinol-5-sulfonic acid was nearly devoid of antimicrobial activity, due to what was believed to be an unfavorable partition coefficient. since twenty six 8-quinolinol-5- and 7-sulfonic acids were available from our previous work, they were tested against six fungi. the 7-chloro and 7-bromo-5-sulfonic acids and the 5-chloro and 5-bromo-7-sulfonic acids showed fungal inhibition within one order of magnitude of that of 8-quinolinol. it is suggested that a nonchelating mechanism is in part res ...200212650598
altering the expression of two chitin synthase genes differentially affects the growth and morphology of aspergillus oryzae.in aspergillus oryzae, one full-length chitin synthase (chsb) and fragments of two other chitin synthases (csma and chsc) were identified. the deduced amino acid sequence of chsb was similar (87% identity) to chsb from aspergillus nidulans, which encodes a class iii chitin synthase. the sequence obtained for csma indicated that it had high similarity to class v chitin synthases. chsb and csma disruption strains and a strain in which chsb transcription was controlled were constructed using the ni ...200212480906
protein expression and secretion in the yeast yarrowia lipolytica.strains and vectors for protein expression and secretion have been developed in the yeast yarrowia lipolytica. host strains were constructed with non-reverting auxotrophic markers, deletions of protease-encoding genes, and carrying a docking platform. to drive transcription, either the synthetic hp4d or the inducible pox2 promoter were used. protein secretion is either directed by the targeting sequence of the alkaline extracellular protease or the extracellular lipase (lip2p) signal sequence. w ...200212702287
aspergillus oryzae in solid-state and submerged fermentations. progress report on a multi-disciplinary project.we report the progress of a multi-disciplinary research project on solid-state fermentation (ssf) of the filamentous fungus aspergillus oryzae. the molecular and physiological aspects of the fungus in submerged fermentation (smf) and ssf are compared and we observe a number of differences correlated with the different growth conditions. first, the aerial hyphae which occur only in ssfs are mainly responsible for oxygen uptake. second, ssf is characterised by gradients in temperature, water activ ...200212702312
telomeric repeat sequence of aspergillus oryzae consists of dodeca-nucleotides.four telomeres in the chromosomes of aspergillus oryzae nfri1599 were cloned and sequenced. the telomeric repeat sequence of a. oryzae consisted of dodeca-nucleotides: ttagggtcaaca. the length of the telomeric repeat tract was 114-136 bp, which corresponds to 9-11 repeats of the dodeca-nucleotide sequence. compared to a chromosome internal control (18s rdna), the telomeric sequences were found to be sensitive to bal31 exonuclease digestion, thus proving that the identified telomeric repeat seque ...200312698283
cloning and overexpression of beta-n-acetylglucosaminidase encoding gene naga from aspergillus oryzae and enzyme-catalyzed synthesis of human milk oligosaccharide.we isolated a beta-n-acetylglucosaminidase encoding gene from the filamentous fungus aspergillus oryzae, and designated it naga. the naga gene encoded a polypeptide of 600 amino acids with significant similarity to glucosaminidases and hexosaminidases of various eukaryotes. a. oryzae strain carrying the naga gene under the control of the improved glaa promoter produced large amounts of beta-n-acetylglucosaminidase in a wheat bran solid culture. the beta-n-acetylglucosaminidase was purified from ...200312723619
substrate specificities of deuterolysin from aspergillus oryzae and electron paramagnetic resonance measurement of cobalt-substituted deuterolysin.the substrate specificities of deuterolysin, a 19-kda zinc-protease (ec 3.4.24.39) from aspergillus oryzae, were investigated at ph 9.0 with various fluorogenic acyl-peptide-4-methylcoumaryl-7-amides (peptide-mcas). n-butoxycarbonyl-arg-val-arg-arg-mca was the best substrate for deuterolysin. we therefore measured its kinetic parameters. deuterolysin had high activity toward the peptide bonds next to pairs of basic residues in calf thymus histone h4. the specificity of cobalt-substituted deutero ...200312728984
calotropis procera (sodom apple)--a potential material for enzyme purification.a simple method based on precipitation with calotropis procera latex was developed for the purification of crude enzyme from fermentation broth. c. procera latex (10(-2) dilution) clarified and concentrated the crude amylase of aspergillus oryzae 4-fold with 97% recovery of the initial amylase activity in the filtrate in a single step operation. the latex was stable at ph < or = 4.5 and there was no significant difference (p < or = 0.05) in the purification potential of the latex at 4 and 28 deg ...200312733587
molecular characterisation and expression analysis of the first hemicellulase gene (bxl1) encoding beta-xylosidase from the thermophilic fungus talaromyces emersonii.the gene coding for beta-xylosidase, bxl1, has been cloned from the thermophilic filamentous fungus, talaromyces emersonii. this is the first report of a hemicellulase gene from this novel source. at the genomic level, bxl1 consists of an open reading frame of 2388 nucleotides with no introns that encodes a putative protein of 796 amino acids. the bxl1 translation product contains a signal peptide of 21 amino acids that yields a mature protein of 775 amino acids, with a predicted molecular mass ...200312763033
gene cloning and functional analysis of a second delta 6-fatty acid desaturase from an arachidonic acid-producing mortierella fungus.we demonstrated that mortierella alpina 1s-4 has two delta 6-desaturases, which are involved in the desaturation of linoleic acid to gamma-linolenic acid. for one of the two delta 6-desaturases, designated as delta 6i, gene cloning and its heterologous expression in a fungus, aspergillus oryzae, has previously been reported. in addition, we indicated in this paper that there is an isozyme of the two delta 6-desaturases, designated as delta 6ii, in m. alpina 1s-4. the predicted amino acid sequenc ...200312784608
functional analysis of the calcineurin-encoding gene cnaa from aspergillus oryzae: evidence for its putative role in stress adaptation.the presence of putative stre (stress response regulatory element) and hsf (heat-shock factor) transcription factor binding sites in the promoter region of the gene encoding calcineurin ( cnaa) from aspergillus oryzae implicated a probable role for calcineurin in the stress response. the activity of calcineurin was enhanced during growth of the wild-type strain in the presence of 1 m nacl (2.6-fold), at alkaline ph 10.0 (2.9-fold) and at 37 degrees c (1.6-fold). the induction of cnaa antisense e ...200312709783
preventive effect of fermented brown rice and rice bran on diethylnitrosoamine and phenobarbital-induced hepatocarcinogenesis in male f344 rats.epidemiological and preclinical studies have suggested that nutrition plays an important role in the etiology of cancer. our group previously demonstrated that rice germ or fermented brown rice has a preventive effect on colorectal carcinogenesis. the experiment described here was examined for the potential anticancer properties of brown rice fermented by aspergillus oryzae (fbra) in male f344 rats using inhibition of diethylnitrosoamine (den) and phenobarbital (pb)-induced hepatocarcinogenesis ...200312792738
pulsed feeding during fed-batch aspergillus oryzae fermentation leads to improved oxygen mass transfer.productivity in many fungal fermentations is detrimentally affected by high broth viscosity and consequent reduced oxygen mass transfer capacity. the goal here was to determine whether pulsed feeding of limiting carbon in a fungal fermentation could lead to reduced viscosity and improved oxygen mass transfer. as a model, an industrially relevant recombinant strain of aspergillus oryzae was grown in carbon-limited, fed-batch mode. maltodextrin was used as a carbon source and was added either cont ...200312790687
substrate aggregation due to aerial hyphae during discontinuously mixed solid-state fermentation with aspergillus oryzae: experiments and modeling.solid-state fermentation (ssf) is prone to process failure due to channeling caused by evaporative cooling and the formation of an interparticle mycelium network. mixing is needed to break the mycelium network and to avoid such failure. this study presents the first attempt to quantify and predict the effect of mycelium bonds on particle mixing and vice versa. we developed a novel experimental set-up to measure the tensile strength of hyphal bonds in ssf: aspergillus oryzae was cultivated betwee ...200312827692
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