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
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
a transcriptional activator, aoxlnr, controls the expression of genes encoding xylanolytic enzymes in aspergillus 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
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
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
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
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
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
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
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
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
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
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
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
ultra-high vacuum and microorganisms.experimental study of the space environment effect on living organisms is one of the most important tasks of exobiology. however location of the living organisms under study outside space vehicles and sputniks is combined with technical difficulties. therefore it is much more suitable to reproduce conditions found in space in laboratory and to study its effect on living cells. it was proved that low temperatures, even those approaching absolute zero do not kill microorganisms. the fact of microo ...196512035798
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
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
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
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
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
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 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
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
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
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
[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
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
subtractive cloning of cdna from aspergillus oryzae differentially regulated between solid-state culture and liquid (submerged) 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
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
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
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
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
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
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
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
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
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 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
kinetic analysis of enhanced thermal stability of an alkaline protease with engineered twin disulfide bridges and calcium-dependent stability.the thermal stability of a cysteine-free alkaline protease (alp) secreted by the eukaryote aspergillus oryzae was improved both by the introduction of engineered twin disulfide bridges (cys-69/cys-101 and cys-169/cys-200), newly constructed as part of this study, and by the addition of calcium ions. we performed an extensive kinetic analysis of the increased thermal stability of the mutants as well as the role of calcium dependence. the thermodynamic activation parameters for irreversible therma ...200312451555
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
pulsed feeding during fed-batch fungal fermentation leads to reduced viscosity without detrimentally affecting protein expression.the goal in this study was to determine if pulsed addition of substrate could be used to alter filamentous fungal morphology during fermentation, to result in reduced broth viscosity. in all experiments, an industrially relevant strain of aspergillus oryzae was grown in 20-liter fermentors. as a control, cultures were fed limiting substrate (glucose) continuously. tests were performed by altering the feeding strategy so that the same total amount of glucose was fed in repeated 300-s cycles, with ...200312474257
altering the expression of two chitin synthase genes differentially affects the growth and morphology of aspergillus 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
molecular breeding of yeast with higher metal-adsorption capacity by expression of histidine-repeat insertion in the protein anchored to the cell wall.a fusion protein of hexa-histidine repeat (his) and glycosylphosphatidylinositol (gpi)-anchor region of saccharomyces cerevisiae cwp1 with aspergillus oryzae taka-amylase a (taa) was expressed on the yeast cell surface. the expressed fusion protein (taa-his-cwp1) was localized on the cell wall and demonstrated amylolytic activity. in comparison with the taa-cwp1 expressing strain, these cells exhibited 1.6- to 2.8-fold higher adsorbing capacity for cu(2+), ni(2+), and zn(2+).200012483584
production of alkaline protease by a genetically engineered aspergillus oryzae u1521.the production of alkaline protease of aspergillus oryzae u1521 was examined in liquid culture. in a culture of defatted soybean only, it gave satisfactory enzyme yields at 584,000 u/g defatted soybean. when various carbohydrates were supplemented, enzyme production was significantly increased. an increase in production by lactose was the most marked. enrichment with casitone or casein increased productivity, but not cornsteep solid. media formulation (g/l) of defatted soybean 10, lactose 5, cas ...199912501379
evolutionary relationships among aspergillus oryzae and related species based on the sequences of 18s rrna genes and internal transcribed spacers. 199812501432
effect of deletion of chitin synthase genes on mycelial morphology and culture viscosity in aspergillus oryzae.the objective of this study was to quantify the effect of disrupting two chitin synthases, chsb and csma, on the morphology and rheology during batch cultivation of aspergillus oryzae. the rheological properties were characterized in batch cultivations at different biomass concentrations (from 3.4-22.5 g kg(-1) biomass) and the power-law model adequately described the rheological properties. in the cultivations there were pellets, clumps, and freely dispersed hyphal elements. the different morph ...200312514801
aorsin, a novel serine proteinase with trypsin-like specificity at acidic ph.a proteinase that hydrolyses clupeine and salmine at acidic ph, called aorsin, was found in the fungus aspergillus oryzae. purified aorsin also hydrolysed benzyloxycarbonyl-arg-arg-4-methylcoumaryl-7-amide optimally at ph 4.0. the specificity of aorsin appeared to require a basic residue at the p(1) position and to prefer paired basic residues. aorsin activated plasminogen and converted trypsinogen to trypsin. the trypsin-like activity was inhibited strongly by antipain or leupeptin, but was not ...200312519073
dipeptidyl peptidase iv on activated t cells as a target molecule for therapy of rheumatoid arthritis.the extracellular domain of the t cell co-stimulatory molecule cd26 possesses dipeptidyl peptidase iv (dp iv) enzyme activity. activated t cells are known to increase expression of cell surface dp iv and some specific inhibitors of this enzyme have been reported to suppress t cell function. previously we have identified a dp iv inhibitor, designated tmc-2, found in culture supernatant of aspergillus oryzae. administration of tmc-2 to rats with adjuvant arthritis caused marked suppression of paw ...200312519388
change in maltose- and soluble starch-hydrolyzing activities of chimeric alpha-glucosidases of mucor javanicus and aspergillus oryzae.the chimeric alpha-glucosidases of mucor javanicus and aspergillus oryzae, which has high activity toward not only maltooligosaccharides but also soluble starch and has high activity toward maltooligosaccharides but faint activity toward soluble starch, respectively, were constructed by shuffling the c-terminal regions where low homology is observed between the two enzymes. the chimera genes were expressed in pichia pastoris to produce and secrete the enzymes that have predicted molecular masses ...200312535604
sequence comparison of aflr from different aspergillus species provides evidence for variability in regulation of aflatoxin production.aflatoxin contamination of foods and feeds is a world-wide agricultural problem. aflatoxin production requires expression of the biosynthetic pathway regulatory gene, aflr, which encodes a cys6zn2-type dna-binding protein. homologs of aflr from aspergillus nomius, bombycis, parasiticus, flavus, and pseudotamarii were compared to investigate the molecular basis for variation among aflatoxin-producing taxa in the regulation of aflatoxin production. variability was found in putative promoter consen ...200312553937
[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
pulsed addition of limiting-carbon during aspergillus oryzae fermentation leads to improved productivity of a recombinant enzyme.fungal morphology in many filamentous fungal fermentations leads to high broth viscosity which limits oxygen mass transfer, and often results in reduced productivity. the objective in this study was to determine if a simple, fed-batch, process strategy-pulsed addition of limiting-carbon source-could be used to reduce fungal broth viscosity, and increase productivity of an industrially relevant recombinant enzyme (glucoamylase). as a control, three aspergillus oryzae fed-batch fermentations were ...200312569630
single-strand-specific nucleases.single-strand-specific nucleases are multifunctional enzymes and widespread in distribution. their ability to act selectively on single-stranded nucleic acids and single-stranded regions in double-stranded nucleic acids has led to their extensive application as probes for the structural determination of nucleic acids. intracellularly, they have been implicated in recombination, repair and replication, whereas extracellular enzymes have a role in nutrition. although more than 30 single-strand-spe ...200312586391
crystal structures of aspergillus oryzae aspartic proteinase and its complex with an inhibitor pepstatin at 1.9a resolution.the x-ray structures of aspergillus oryzae aspartic proteinase (aoap) and its complex with inhibitor pepstatin have been determined at 1.9a resolution. aoap was crystallized in an orthorhombic system with the space group p2(1)2(1)2(1) and cell dimensions of a=49.4a, b=79.4a, and c=93.6a. by the soaking of pepstatin, crystals are transformed into a monoclinic system with the space group c2 and cell dimensions of a=106.8a, b=38.6a, c=78.7a, and beta=120.3 degrees. the structures of aoap and aoap/p ...200312595261
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
separation of enzymes from polyenzyme mixture used in medicine and pharmacy. ii. purification and characterization of extracellular beta-glycosidases with high transglycosylation activities from aspergillus oryzae.three extracellular beta-glycosidases with different substrate specificities have been isolated from aspergillus oryzae (luizym) and purified to electrophoretic homogeneity by molecular-sieve and ion-exchange chromatographic methods. the enzymes were characterized as monomeric glycoproteins with an estimated molecular mass of 95 kda by sds-page and 92 kda by gel-permeation chromatography on superose 12 hr 10/30. beta-glycosidase i (phopt 4.8; topt 40 degrees c, pl 4.5) was able to catalyze the h ...200312622257
solubilization of trichloroacetic acid (tca) precipitated microbial proteins via naoh for two-dimensional preparing intracellular microbial samples for one- or two-dimensional electrophoresis, trichloroacetic acid (tca) precipitation is frequently used to remove interfering compounds. solubilization of tca precipitate typically requires the addition of a number of chaotropes or detergents, in a multistep process, that requires hours to carry out. in this study, a simple, rapid, one-step method to solubilize tca precipitated proteins is presented. precipitated proteins are pretreated with 0.2 m na ...200312643547
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
mycotoxin production in wheat grains by different aspergilli in relation to different relative humidities and storage periods.four different aspergilli (aspergillus oryzae, a. parasiticus, a. terreus and a. versicolor) were grown on wheat grains underdifferent degrees of relative humidity 14, 50, 74, 80 and 90%. samples of wheat grains were taken monthly for a period of six months and examined for mycotoxin production. a. oryzae was found to produce aflatoxins b1, b2, zearalenone, don and t-2 toxins under elevated degrees of humidity and prolonged periods of storage. a. parasiticus produced aflatoxins b1, g1, niv, don ...200312653428
electrophoretically mediated reaction of glycosidases at a nanoliter scale.we have investigated electrophoretically mediated microanalysis (emma) for the assay of a native glyco-enzyme. as a representative of this class of enzyme, beta-glucosidase was selected, and the reaction was analyzed. our emma was based on the plug-plug interaction of enzyme and substrate plugs, which is essential to reduce quantities of materials. furthermore, we have addressed the problem of incompatibility of the enzymatic reaction and separation of the reactants. as a result, emma of native ...200312658703
phya gene product of aspergillus ficuum and peniophora lycii produces dissimilar phytases.phya gene products of aspergillus ficuum (af) and peniophora lycii (pl) as expressed in industrial strains of aspergillus niger and aspergillus oryzae, respectively, were purified to homogeneity and then characterized for both physical and biochemical properties. the pl phytase is 26 amino acid residues shorter than the af phytase. dynamic light scattering studies indicate that the active af phytase is a monomer while the pl phytase is a dimer. while both of the phytases retained four identical ...200312659840
inducive effect of cresoquinone on microbiological transformation of l-tyrosine to 3,4 dihydroxy phenyl l-alanine by aspergillus oryzae ng-11(p1).the present work describes the inducive effect of cresoquinone on microbiological transformation of l-tyrosine to 3,4 dihydroxy phenyl l-alanine ( l-dopa) by aspergillus oryzae ng-11(p1). mould mycelium was used for biochemical conversion of l-tyrosine to l-dopa because tyrosinases, beta-carboxylases and tyrosine hydroxylases are intracellular enzymes. the maximum conversion of l-tyrosine to l-dopa (0.428 mg/ml) was achieved after 60 min of biochemical reaction. to enhance the production of l-do ...200312664148
enzymatic preparation of ginsenosides rg2, rh1, and f1.during investigation of the hydrolysis of a protopanaxatriol-type saponin mixture by various glycoside hydrolases, crude preparations of beta-galactosidase from aspergillus oryzae and lactase from penicillium sp. were found to produce two minor saponins, ginsenoside rg(2) [6-o-(alpha-l-rhamnopyranosyl-(1-->2)-beta-d-glucopyranosyl)-20(s)-protopanaxatriol] and ginsenoside rh(1) (6-o-beta-d-glucopyranosyl-20(s)-protopanaxatriol), respectively, in high yields. moreover, a naringinase preparation fr ...200312672992
enzymatic preparation of ginsenosides rg2, rh1, and f1 from protopanaxatriol-type ginseng saponin mixture.during investigations on the hydrolysis of a protopanaxatriol-type saponin mixture by various glycoside hydrolases, it was found that two minor saponins, ginsenosides rg 2 and rh 1, were formed in high yields by crude beta-galactosidase from aspergillus oryzae and crude lactase from penicillium sp., respectively. moreover, a crude preparation of naringinase from penicillium decumbens readily hydrolyzed a protopanaxatriol-type saponin mixture to give an intestinal bacterial metabolite, ginsenosid ...200312677539
fungal beta-n-acetylhexosaminidases with high beta-n-acetylgalactosaminidase activity and their use for synthesis of beta-galnac-containing oligosaccharides.about 60 fungal strains were tested for production of extracellular beta-n-acetylhexosaminidases. a unique beta-n-acetylhexosaminidase with the beta-galnac-ase/beta-glcnac-ase ratio of 2.3-2.8 was found in the culture filtrates of some strains of penicillium oxalicum. addition of 20% (w/v) mgso(4) increased the beta-galnac-ase/beta-glcnac-ase ratio to the value of 3.35. cultivation conditions influence this ratio as well. beta-n-acetylhexosaminidases from p. oxalicum ccf 2430 and aspergillus ory ...200312681926
characterization of aspergillus oryzae fermentation extract effects on the rumen fungus neocallimastix frontalis, eb 188. part 1. zoospore development and physiology.experiments were performed to determine the effect of aspergillus oryzae (ao) fermentation extract on zoospore development in the rumen fungus neocallimastix frontalis eb 188. powdered product, or liquid extract prepared from such powder, was added at the recommended value for supplementation in dairy cattle. stationary and stirred cultures were periodically sampled and assayed for extracellular and intracellular protein and enzymes, gas production, zoospore production and maturation, and carbon ...200412690417
characterization of aspergillus oryzae fermentation extract effects on the rumen fungus neocallimastix frontalis, eb 188. part 2. carbon source utilization and effects on zoospore production.the effect of a commercial aspergillus oryzae fermentation extract on the utilization of carbon source and zoospore production by the rumen fungus neocallimastix frontalis eb 188 was determined. in addition, the composition of a soluble extract prepared from the commercial product was analyzed. this extract was added to n. frontalis eb 188 cultures grown on a variety of substrates and periodically assayed for protein, enzymes, zoospore production, and carbon source utilization. the powdered prod ...200412690418
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
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
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
[quantitative determination of 3-nitropropionic acid by hplc].a reversed-phase liquid chromatographic method to determine the concentration of 3-nitropropionic acid (3-npa) in the culture medium with aspergillus oryzae was developed. the culture medium samples were extracted with ethyl acetate. the analytic column was hypersil c18 and the mobile phase was ch3cn/kh2po4(0.02 mol/l, ph 3.0) (1/3, v/v). the sample was detected at uv210 nm. the detection limit of this method was at 10 micrograms/kg level and recoveries were 90.1%-98.0.199912712703
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 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
in vivo visualization of the distribution of a secretory protein in aspergillus oryzae hyphae using the rnta-egfp fusion protein.a fusion gene encoding ribonuclease t1-egfp (rnta-egfp) was constructed and expressed to use it as a tool for studies on the secretory pathway in aspergillus oryzae. the successful secretion of the intact rnta-egfp fusion protein was detected by fluorescence measurement and western analysis. with use of the rnta-egfp system, we were able to see high fluorescence at hyphal tips and observe concentrated fluorescence at septa in basal cells during growth at optimal conditions. cold or heat shock du ...200312729022
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
a novel heterofunctional epoxy-amino sepabeads for a new enzyme immobilization protocol: immobilization-stabilization of beta-galactosidase from aspergillus oryzae.the properties of a new and commercially available amino-epoxy support (amino-epoxy-sepabeads) have been compared to conventional epoxy supports to immobilize enzymes, using the beta-galactosidase from aspergillus oryzae as a model enzyme. the new support has a layer of epoxy groups over a layer of ethylenediamine that is covalently bound to the support. this support has both a great anionic exchanger strength and a high density of epoxy groups. epoxy supports require the physical adsorption of ...200312790680
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
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
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
construction of an efficient expression system for aspergillus isopullulanase in pichia pastoris, and a simple purification method.aspergillus niger atcc 9642 isopullulanase (ipu) was heterologously expressed by pichia pastoris gs115 under three different signal sequences of saccharomyces cerevisiae acid phosphatase, s. cerevisiae alpha-factor prepro peptide, and a. niger isopullulanase. one-step purification using lectin con a affinity chromatography yielded recombinant ipu (ipu-pp) with high purity. ipu-pp had a higher carbohydrate content than native ipu and ipu-ao expressed in a. oryzae m-2-3. ipu-pp hydrolyzed various ...200312834298
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