cloning and characterization of the bipa gene encoding er chaperone bip from aspergillus oryzae.we describe the cloning and characterization of the er localized chaperone (bip) encoding gene from the filamentous fungus aspergillus oryzae. the bip encoding gene, designated bipa, has three introns and encodes a protein with 672 amino acids, which has a high homology with various bip-like proteins. sequences resembling heat shock elements (hses) and unfolded protein response (upr) elements, as found in the saccharomyces cerevisiae kar2 promoter, are present in the 5' no coding region of the b ...199916232647
production and some properties of salt-tolerant beta-xylosidases from a shoyu koji mold, aspergillus oryzae in solid and liquid cultures.beta-xylosidase production from a shoyu (soy sauce) koji mold, aspergillus oryzae hl15, cultured in solid and liquid media was examined and some properties of the enzymes were studied. three beta-xylosidases (xy11, xy12 and xy13) were easily extracted with 0.5% nacl from a solid medium and purified homogeneously on sds-page by chromatography. on the other hand, in a liquid medium, a. oryzae hl15 produced mainly cell-wall-bound beta-xylosidases which could not be extracted with 0.5% nacl or any d ...199916232648
molecular cloning of the isoamyl alcohol oxidase-encoding gene (mrea) from aspergillus oryzae.isoamyl alcohol oxidase (iaaod) is a novel enzyme that catalyzes the formation of isovaleraldehyde, which is the main component of mureka that gives sake an off-flavor (yamashita et al. biosci. biotechnol. biochem., 63, 1216-1222, 1999). we cloned the genomic dna sequence encoding iaaod from a koji mold, aspergillus oryzae, using a pcr-amplified dna fragment corresponding to the partial amino acid sequences of the purified protein as a probe. the cloned gene comprises 1903 bp of an open reading ...200016232791
production and product quality assessment of human hepatitis b virus pre-s2 antigen in submerged and solid-state cultures of aspergillus oryzae.pre-s2 is a diagnostically important antigen of human hepatitis b virus (hbv). in order to produce pre-s2 antigen in aspergillus oryzae, the gene [pre-s2]3, which encodes a tandemly triplicated repeat of pre-s2 polypeptides was fused with the partial glaa gene encoding glucoamylase lacking the starch-binding domain. in submerged culture, a. oryzae transformants carrying glaa-[pre-s2]3 secreted a heterogeneously glycosylated form of the fusion protein that was partially degraded. contrarily, util ...200016232829
accumulation of a recombinant aspergillus oryzae lipase artificially localized on the bacillus subtilis cell surface.cutl cdna encoding an extracellular lipase, l1, from aspergillus oryzae was fused to the cell wall-binding domain (cwb) region of a plasmid, phcb3r. sds-polyacrylamide gel electrophoresis (page) and zymography of proteins extracted from the cell surface of bacillus subtilis 168 harboring a fused lipase plasmid (phcb3rcl) revealed that the fused gene product, cwb-cutl, was localized in the b. subtilis cell wall and retained lipase activity. b. subtilis wasd (wpra sigd), recently used for the accu ...200016232883
rapid detection of homologously integrated dna fragments and accurate quantitation of their copy number in transgenic aspergillus oryzae by pcr.a simple and rapid method for the analysis of artificially introduced dna fragments has been developed using competitive pcr and long and accurate pcr. the locus and the copy number of the dna fragments in each aspergillus oryzae transformant could be detected more rapidly and accurately by this method than by the conventional southern hybridization method.200016232915
improvement of production of kojic acid by a mutant strain aspergillus oryzae, mk107-39.a strain designated mk107-39, producing kojic acid with a high yield, was obtained by a new screening method using a 96-well microtiter plate after ntg treatment of aspergillus oryze atcc 22788. the amount of kojic acid produced by strain mk107-39 in a shaking flask was 28 g/l from 100 g/l of glucose, which was 7.7-times higher than that produced by parent strain. the kojic acid yields per cell and the amount of glucose consumed were 9.8 and 6.0-times higher than those of the parent strain. base ...200116232988
a second pectin lyase gene (pel2) from aspergillus oryzae kbn616: its sequence analysis and overexpression, and characterization of the gene products.a second pectin lyase gene, designated pel2, was isolated from a shoyu koji mold aspergillus oryzae kbn616 and characterized. the structural gene comprised 1306 bp with three introns. the orf encoded 375 amino acids with a signal peptide of 19 amino acids. the deduced amino acid sequence showed high similarity to those of a. oryzae pel1, aspergillus niger pectin lyases and glomerella cingulata pn1a. the pel2 gene was overexpressed under the control of the promoter of the a. oryzae tef1 gene for ...200116233008
overexpression and purification of aspergillus aculeatus beta-mannosidase and analysis of the integrated gene in aspergillus expression plasmid for the manb gene encoding aspergillus aculeatus beta-d-mannosidase (manb) was constructed by using an expression vector carrying an improved promoter. after transformation of a. oryzae by the plasmid, several transformants formed colonies emitting fluorescence on a plate containing 4-methylumbelliferyl beta-d-mannopyranoside (mu-man) under uv-irradiation. the transformant that displayed the strongest fluorescence, named a. oryzae bmn1, produced about 270 mg manb/l in liqui ...200116233072
kojic acid production in an airlift bioreactor using partially hydrolyzed raw corn a culture of aspergillus oryzae mk-107-39 in a 3-l airlift bioreactor, kojic acid was not produced when glucose/wheat germ medium (gm1) was used. however, when a jar fermentor was used, the kojic acid yield was high. a suitable medium for culture in an airlift bioreactor consisting of partially hydrolyzed corn starch and a small amount of corn steep liquor (csl) (sm1) was selected. in the cultivation in the airlift bioreactor using sm1, nearly 40 g/l of kojic acid was produced, which was the ...200116233111
efficient production of a heterologous peroxidase, dyp from geotrichum candidum dec 1, on solid-state culture of aspergillus oryzae rd005.the productivity of a peroxidase (dyp) originating from geotrichum candidum dec 1 was enhanced in the solid-state culture using aspergillus oryzae rd005. when the humidity, water content, and temperature were adjusted to 60%, 50% and 27 degrees c, respectively, the productivity of dyp reached 5.3 g per kilogram wheat bran, which was used as the solid medium. the yield of 5.3 g per kg wheat bran corresponded to the yield of a 56 kg submerged culture. the productivity per gram carbon of the medium ...200116233153
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
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
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
molecular breeding of the mureka-non-forming sake koji mold from aspergillus oryzae by the disruption of the mrea gene.mureka-non-forming sake koji molds were constructed from an aspergillus oryzae industrial strain by the disruption of the mrea gene using a host-vector system with the ptra gene as a dominant selectable marker. all of the mrea gene disruptants obtained retained the advantages of the host strain in terms of the brewing characteristics, while their isoamyl alcohol oxidase (iaaod) activities were significantly lower than that of the host strain. sake brewing was successfully carried out using the k ...200316233364
dffa gene from aspergillus oryzae encodes l-ornithine n5-oxygenase and is indispensable for deferriferrichrysin biosynthesis.we identified and analyzed the dffa gene from aspergillus oryzae which encodes l-ornithine n5-oxygenase involved in the biosynthesis of deferriferrichrysin, a type of siderophore which is a low-molecular-weight iron chelating compound. from among more than 20,000 clones in an a. oryzae est (expressed sequence tag) library, we found only one clone encoding a protein that exhibited homology to theustilago maydis sid1 protein (sid1) and pseudomonas aeruginosa pvda protein (pvda), both known as the ...200316233371
synergistic degradation of arabinoxylan with alpha-l-arabinofuranosidase, xylanase and beta-xylosidase from soy sauce koji mold, aspergillus oryzae, in high salt condition.this study addresses induction and some properties of alpha-l-arabinofuranosidase from a soy sauce koji mold, aspergillus oryzae hl15, cultured on solid and liquid media. alpha-l-arabinofuranosidase was induced by soybean polysaccharide and secreted into media on solid cultivation; the enzyme was associated with mycelium as a cell-wall-bound form in liquid cultivation. a major alpha-l-arabinofuranosidase, which was purified homogeneously on sds-page, showed highest activity in the presence of 10 ...200316233386
production and properties of phytase and acid phosphatase from a sake koji mold, aspergillus oryzae.we identified three types of acid phosphatase (acp-i, acp-ii, and acp-iii) produced by aspergillus oryzae in a submerged culture using only phytic acid as the phosphorous substrate. the optimum ph for the activities of the three enzymes was in the range of 4.5 to 5.5. analysis of the substrate specificities of these enzymes revealed that acp-i and acp-iii were acid phosphatases, and acp-ii was a phytase. these enzymes were produced during different periods of mycelial growth: acp-ii was produced ...200316233418
production of two types of phytase from aspergillus oryzae during industrial koji our previous study, it was determined that phytase produced by aspergillus oryzae plays an important role in supplying phosphate to yeast in the process of making sake. during koji making, two types of phytase (phy-i and phy-ii) are produced. the purified phytases have high thermal and ph stability, in comparison to phytase purified from a submerged culture (acp-ii). in the present study, phy-i and phy-ii retained their activities for 45 h. the nh2-terminal sequence of phy-1, which is eight a ...200316233440
overexpression of aspergillus aculeatus cellobiohydrolase i in aspergillus express the cbhi gene, encoding aspergillus aculeatus cellobiohydrolase i (cbhi), in aspergillus oryzae, a plasmid was constructed. the strain that displayed the strongest cbhi activity among the transformants produced about 941 mg/l in liquid culture. it was confirmed by a pcr method that the plasmid was integrated at the niad locus.200316233469
growth of aspergillus oryzae during treatment of cassava starch processing wastewater with high content of suspended solids.aspergillus oryzae ifo 30113 was used for the treatment of the cassava starch processing (csp) wastewater. the observations on the fungal morphology showed that, in the shake flasks containing the csp wastewater with the high concentration of suspended solids, the formation of pellets originated from the adherence of germinated spores to solid particles in medium. the attached solid particles were also digested during the fungal fermentation and resulted in the formation of the smooth and hollow ...200416233638
cloning of a novel tyrosinase-encoding gene (melb) from aspergillus oryzae and its overexpression in solid-state culture (rice koji).we have cloned a novel tyrosinase-encoding gene (melb) specifically expressed in solid-state culture of aspergillus oryzae. a tyrosinase-encoding gene (melo) from a. oryzae was already cloned and the protein structures of its catalytic and copper binding domains were investigated. however, our recent results revealed that the melo gene was highly expressed in submerged culture but not in solid-state culture. because tyrosinase activity was also detected in solid-state culture, we assumed that an ...200416233650
cloning and expression of a nad+-dependent xylitol dehydrogenase gene (xdha) of aspergillus oryzae.xdha, which encodes a xylitol dehydrogenase gene, was cloned from aspergillus oryzae genomic dna. it consists of 1214 bp structural region, which is interrupted by two introns, and encodes 358-amino-acid protein (38,197 da). it is similar to the known nad(+)-dependent xylitol dehydrogenase (ec the gene was expressed in escherichia coli bl21-ai using a t7 promoter. the cell-free extract of the transformant showed a 36.5 kda band upon sds-page and nad(+)-dependent xylitol dehydrogenase a ...200416233653
isolation and characterization of a novel gene encoding alpha-l-arabinofuranosidase from aspergillus oryzae.we cloned and characterized a novel gene (abfa) encoding alpha-l-arabinofuranosidase (alpha-l-afase) from aspergillus oryzae. one clone homologous to the alpha-l-afase gene of thermotoga maritima was found in an expressed sequence tag (est) library of a. oryzae and a corresponding gene was isolated. molecular analysis showed that the abfa gene carried six exons interrupted by five introns and had an open reading frame encoding 481 amino acid residues. the amino acid sequence similarity at active ...200416233670
characteristics of alpha-glucosidase production from recombinant aspergillus oryzae by membrane-surface liquid culture in comparison with various cultivation methods.alpha-glucosidase was produced using recombinant aspergillus oryzae by membrane-surface liquid culture (mslc), a method previously developed by the authors and the results compared with other methods, including shaking flask culture (sfc), agar-plate culture (apc), culture on urethane sponge supports (usc), and liquid surface culture (lsc) to determine possible reasons for the advantageous features of mslc. when yeast extract was used as a nitrogen source, the amount of enzyme produced by mslc w ...200416233690
a novel amine oxidase-encoding gene from aspergillus oryzae.we cloned a novel gene (aoxa) encoding amine oxidase (aox) from aspergillus oryzae. one cdna clone showing extreme homology to the aox-encoding genes was found in an expressed sequence tag (est) library of a. oryzae. molecular analysis revealed that the aoxa carried four exons interrupted by three introns and had an open reading frame encoding 672 amino acid residues. the deduced amino acid sequence showed about 83.5% identity to the aspergillus niger ao-i. the strictly conserved residues for co ...200416233720
cloning and expression analysis of two catalase genes from aspergillus oryzae.fungi contain distinct genes encoding the same class of enzyme that are differentially regulated according to conditions. we cloned two catalase genes, cata and catb, from aspergillus oryzae. the cata gene predicts a 747-amino-acid polypeptide sharing 81% identity with aspergillus fumigatus catalase (cata) and 77% with aspergillus nidulans catalase (cata). the catb gene predicts a 725-amino-acid polypeptide sharing 82% identity with a. fumigatus catalase (catb) and 75% with a. nidulans catalase ...200516233832
olive-mill wastewaters: a promising substrate for microbial lipase production.the present study investigated the valorization of olive-mill wastewater (omw) by its use as a possible growth medium for the microbial production of extra-cellular lipase. to this end, strains of geotrichum candidum (nrrl y-552 and y-553), rhizopus arrhizus (nrrl 2286 and isrim 383), rhizopus oryzae (nrrl 6431), aspergillus oryzae (nrrl 1988 and 495), aspergillus niger (nrrl 334), candida cylindracea (nrrl y-17506) and penicillium citrinum (nrrl 1841 and 3754, isrim 118) were screened. all stra ...200616236495
effects of cooked molasses blocks and fermentation extract or brown seaweed meal inclusion on intake, digestion, and microbial efficiency in steers fed low-quality hay.five ruminally, duodenally, and ileally cannulated steers (376 +/- 8.1 kg of initial bw) were used in a 5 x 5 latin square to evaluate effects of cooked molasses block supplementation and inclusion of fermentation extract (aspergillus oryzae) or brown seaweed meal (ascophyllum nodosum) on intake, site of digestion, and microbial efficiency. diets consisted of switchgrass hay (6.0% cp; dm basis) offered ad libitum, free access to water, and one of three molasses blocks (0.341 kg of dm/d; one-half ...200516282634
l-leucine aminopeptidase production by filamentous aspergillus screen various filamentous fungi belonging to aspergillus spp. producing leucine and methionine aminopeptidases.200516305677
value of bilirubin oxidase and its mutants in the diagnosis of elucidate the significance of the coordination amino acid residues in bilirubin oxidase (bo) and their kinetic characteristics, and evaluate whether bo mutants may serve as better diagnostic agent for hyperbilirubinemia.200516305950
taka-amylase a in the conidia of aspergillus oryzae rib40.a study of taka-amylase a of conidia from aspergillus oryzae rib40 was done. during the research, proteins from conidia and germinated conidia were analyzed using sds-page, 2-d gel electrophoresis, western blot analysis, maldi-tof mass spectrometry, and native-page combined with activity staining of taa. the results showed that taa exists not only in germinated conidia but also in conidia. some bands representing degraded products of taa were detected. conidia, which formed on starch (scya), glu ...200516306682
cloning and characterization of saponin hydrolases from aspergillus oryzae and eupenicillium brefeldianum.we purified saponin hydrolases from aspergillus oryzae pf1224 and eupenicillium brefeldianum pf1226. it was confirmed that the enzymes from a. oryzae pf1224 (sda1) and e. brefeldianum pf1226 (sde1) are glycoproteins with molecular masses of 82 and 90 kda respectively. the deduced amino acid sequences of each enzyme from the cloned genes (sda1 or sde1) showed approximately 50% homology with that of the saponin hydrolase sdn1 from neocosmospora vasinfecta var. vasinfecta pf1225 (ddbj accession no. ...200516306700
cloning and expression of nad+-dependent l-arabinitol 4-dehydrogenase gene (lada) of aspergillus oryzae.a gene of aspergillus oryzae, lada, which encodes l-arabinitol 4-dehydrogenase (ec, and its cdna were cloned in escherichia coli. the gene consisted of a 1209-bp coding region, interrupted by a 59-bp intron, which encoded a 382-amino-acid polypeptide (40,812 da). the protein showed 67% identity to a well-studied l-arabinitol 4-dehydrogenase (lad1) of hypocrea jecorina. the cell-free extract of e. coli, which expressed lada cdna, showed l-arabinitol dehydrogenase activity with nad+. it ...200516310740
isolation and analysis of molds from soy sauce koji in thailand.five different isolates of aspergillus and one of mucor were compared with a japanese commercial strain of aspergillus oryzae for proteolytic activity on wheat bran substrate. one isolate of aspergillus with superior protease production, identified as aspergillus flavus var. columnaris, showed no detectable aflatoxin production on glutinous rice or soybean substrate. preliminary tests using this fungus as a koji mold in a traditionally operated factory resulted in a soy sauce superior in quality ...198016345516
purification and characterization of extracellular proteinases of aspergillus oryzae.[this corrects the article on p. 507 in vol. 30.].198316346450
conidium germination rate in wild and domesticated yellow-green aspergilli.conidia of domesticated yellow-green aspergilli from strains of aspergillus oryzae (ahlburg) cohn and aspergillus sojae sakaguchi and yamada ex murakami, used in the preparation of koji inoculum, germinate approximately 3 h sooner than conidia produced by related wild species, aspergillus flavus link ex fr. and aspergillus parasiticus speare. there was no consistent relationship between average conidium size and estimated 50% germination time. germination trials were conducted on czapek agar at ...198416346471
genetic transformation of an argb mutant of aspergillus argb mutant of aspergillus oryzae nrrl 492 has been genetically transformed with the aspergillus nidulans argb gene. protoplasts were generated with a combination of novozyme 234 and beta-glucuronidase and regenerated on sucrose-stabilized minimal medium without arginine as described for a. nidulans. a frequency of 5 to 10 transformants per mug of dna was obtained; however, most transformants appeared abortive. the a. nidulans argb gene and vector sequences appeared to be integrated into the ...198816347669
effect of dicarboxylic acids and aspergillus oryzae fermentation extract on lactate uptake by the ruminal bacterium selenomonas ruminantium.the objective of this study was to determine the effects of l-aspartate, fumarate, l-malate, and an aspergillus oryzae fermentation extract (amaferm) on growth on lactate as well as lactate uptake by selenomonas ruminantium hd4. growth of s. ruminantium in medium that contained 2 g of dl-lactate per liter was stimulated approximately twofold by 10 mm l-aspartate, fumarate, or l-malate after 24 h. both l-aspartate and fumarate increased lactate uptake over 4-fold, while l-malate stimulated uptake ...199016348354
in vitro stimulation of forage fiber degradation by ruminal microorganisms with aspergillus oryzae fermentation extract.aspergillus oryzae fermentation extract (amaferm) was evaluated for its ability to influence degradation of brome grass and switchgrass fiber fractions by mixed ruminal microorganisms in vitro. addition of amaferm at a concentration of 0.067 mg/ml, which is approximately the concentration found in the rumen ecosystem (0.06 mg/ml), increased the degradation of brome grass neutral detergent fiber (ndf) by 28% after fermentation for 12 h (p < 0.01), but had no effect after fermentation for 24 or 48 ...199316349057
filamentous fungi with high cytosolic phospholipid transfer activity in the presence of exogenous phospholipid.the phospholipid transfer activity of cell extracts from 15 filamentous fungus strains grown on a medium containing phospholipids as the carbon source was measured by a fluorescence assay. this assay was based on the transfer of pyrene-labeled phosphatidylcholines forming the donor vesicles to acceptor vesicles composed of egg phosphatidylcholines. the highest phosphatidylcholine transfer activity was obtained with cell extracts from aspergillus oryzae. the presence of exogenous phospholipids in ...199416349388
induced autolysis of aspergillus oryzae (a. niger group): iv. carbohydrates.the examination of substances formed during induced autolysis by aspergillus niger was continued in this work, which dealt in particular with carbohydrates. the autolysate contained a large amount of d-glucose (14 to 20% dry wt) and traces of glycolic aldehyde, dihydroxyacetone, ribose, xylose, and fructose. it also contained glycopeptides (about 10% dry wt), which were split from the cell wall during autolysis and which differed from one another in their level of polymerization and their compos ...196216349623
fungi associated with softening of bisulfite-brined cherries.softening of sound, calcium bisulfite-brined cherries was induced fairly quickly by brining them with cherries rotted by aspergillus niger, cytospora leucostoma, and penicillium expansum, but not with cherries rotted by a variety of other microorganisms, including alternaria sp., aspergillus oryzae, aureobasidium pullulans, botrytis cinerea, cladosporium sp., mucor racemosus, rhizopus stolonifer, and sclerotinia fructicola. rapid softening was correlated with the presence of a bisulfite-stable p ...196316349630
utilization of methylthio-s-triazine for growth of soil fungi.aspergillus niger van tieghem, aspergillus tamarii kita, and aspergillus flavus link ex fries utilized the methylthio moiety of 2,4-bis(isopropylamino) -6-methyl-mercapto-s-triazine (prometryne) as a sulfur nutrient source. other soil fungal isolates not affected by prometryne concentrations to 1 mg/ml culture included: aspergillus oryzae (ahlburg) cohn, curvularia lunata (wakker) boedijn, trichoderma viride persoon ex fries, alternaria tenuis nees ex corda, penicillium funiculosum thom, and pae ...197016349873
an iterative type i polyketide synthase pksn catalyzes synthesis of the decaketide alternapyrone with regio-specific octa-methylation.a biosynthetic gene cluster containing five genes, alt1-5, was cloned from alternaria solani, a causal fungus of early blight disease to tomato and potato. homology searching indicated that the alt1, 2, and 3 genes code for cytochrome p450s and the alt4 gene for a fad-dependent oxygenase/oxidase. the alt5 gene encodes a polyketide synthase (pks), named pksn, that was found to be an iterative type i complex reduced-type pks with a c-methyltransferase domain. to identify the pksn function, the alt ...200516356847
sequencing of aspergillus nidulans and comparative analysis with a. fumigatus and a. oryzae.the aspergilli comprise a diverse group of filamentous fungi spanning over 200 million years of evolution. here we report the genome sequence of the model organism aspergillus nidulans, and a comparative study with aspergillus fumigatus, a serious human pathogen, and aspergillus oryzae, used in the production of sake, miso and soy sauce. our analysis of genome structure provided a quantitative evaluation of forces driving long-term eukaryotic genome evolution. it also led to an experimentally va ...200516372000
genome sequencing and analysis of aspergillus oryzae.the genome of aspergillus oryzae, a fungus important for the production of traditional fermented foods and beverages in japan, has been sequenced. the ability to secrete large amounts of proteins and the development of a transformation system have facilitated the use of a. oryzae in modern biotechnology. although both a. oryzae and aspergillus flavus belong to the section flavi of the subgenus circumdati of aspergillus, a. oryzae, unlike a. flavus, does not produce aflatoxin, and its long histor ...200516372010
development of a modified positive selection medium that allows to isolate aspergillus oryzae strains cured of the integrated niad-based plasmid.the nitrate reductase gene (niad) is the most frequently utilized as a selectable marker for homologous integration at the niad locus of aspergillus oryzae. in this study we developed a method for curing of the niad-based plasmid integrated on the a. oryzae genome. positive selection using a modified chlorate medium containing leucine as a nitrogen source enabled efficient isolation of the strains deficient in nitrate assimilation from the niad(+) transformant. pcr analysis of the strains confir ...200516377911
effect of casein hydrolysate, prepared with protease derived from aspergillus oryzae, on subjects with high-normal blood pressure or mild hypertension.casein hydrolysate, prepared with aspergillus oryzae protease, contains angiotensin i-converting enzyme inhibitory peptides, such as val-pro-pro and ile-pro-pro. we conducted a randomized, double-blind, placebo-controlled study to evaluate the effect of casein hydrolysate on the blood pressure of 144 subjects with high-normal blood pressure (n = 104) and mild hypertension (n = 40). subjects were randomly assigned to two groups for a 12-week intake period. in the test group, both systolic (sbp) a ...200516379551
visualization of the endocytic pathway in the filamentous fungus aspergillus oryzae using an egfp-fused plasma membrane protein.endocytosis is an important process for cellular activities. however, in filamentous fungi, the existence of endocytosis has been so far elusive. in this study, we used aouapc-egfp, the fusion protein of a putative uric acid-xanthine permease with enhanced green fluorescent protein (egfp) in aspergillus oryzae, to examine whether the endocytic process occurs or not. upon the addition of ammonium into the medium the fusion protein was internalized from the plasma membrane. the internalization of ...200616380079
micrococcus luteus k-3-type glutaminase from aspergillus oryzae rib40 is salt-tolerant.aspergillus oryzae rib40 possesses the gene of glutaminase (micrococcus luteus k-3-type glutaminase; aogls), which has 40% homology with the salt-tolerant glutaminase from m. luteus k-3 (micrococcus glutaminase). it was found that aogls is a salt-tolerant enzyme, and its properties are similar to those of micrococcus glutaminase.200516384800
comparison of the aflr gene sequences of strains in aspergillus section flavi.aflatoxins are polyketide-derived secondary metabolites produced by aspergillus parasiticus, aspergillus flavus, aspergillus nomius and a few other species. the toxic effects of aflatoxins have adverse consequences for human health and agricultural economics. the aflr gene, a regulatory gene for aflatoxin biosynthesis, encodes a protein containing a zinc-finger dna-binding motif. although aspergillus oryzae and aspergillus sojae, which are used in fermented foods and in ingredient manufacture, h ...200616385126
molecular analysis of an inactive aflatoxin biosynthesis gene cluster in aspergillus oryzae rib help assess the potential for aflatoxin production by aspergillus oryzae, the structure of an aflatoxin biosynthesis gene homolog cluster in a. oryzae rib 40 was analyzed. although most genes in the corresponding cluster exhibited from 97 to 99% similarity to those of aspergillus flavus, three genes shared 93% similarity or less. a 257-bp deletion in the aflt region, a frameshift mutation in nora, and a base pair substitution in vera were found in a. oryzae rib 40. in the aflr promoter, two s ...200616391082
genomics reveals traces of fungal phenylpropanoid-flavonoid metabolic pathway in the f ilamentous fungus aspergillus oryzae.fungal secondary metabolites constitute a wide variety of compounds which either play a vital role in agricultural, pharmaceutical and industrial contexts, or have devastating effects on agriculture, animal and human affairs by virtue of their toxigenicity. owing to their beneficial and deleterious characteristics, these complex compounds and the genes responsible for their synthesis have been the subjects of extensive investigation by microbiologists and pharmacologists. a majority of the funga ...200516410762
studies of the toxicological potential of tripeptides (l-valyl-l-prolyl-l-proline and l-isoleucyl-l-prolyl-l-proline): iii. single- and/or repeated-dose toxicity of tripeptides-containing lactobacillus helveticus-fermented milk powder and casein hydrolysate in rats.the objective of these studies was to assess the toxicological potential of orally administered tripeptides in rats. the studies employed powdered l-valyl-l-prolyl-l-proline (vpp)- and l-isoleucyl-l-prolyl-l-proline (ipp)-containing test articles, including (1) powdered lactobacillus helveticus-fermented milk (fm), (2) pasteurized casein hydrolysate (ch) generated by aspergillus oryzae protease, and (3) synthesized vpp. all test articles were administered by oral gavage to male and female spragu ...200516419576
studies of the toxicological potential of tripeptides (l-valyl-l-prolyl-l-proline and l-isoleucyl-l-prolyl-l-proline): v. a 13-week toxicity study of tripeptides-containing casein hydrolysate in male and female rats.the objective of this multiple-dose toxicity study was to assess the toxicological potential of two tripeptides, l-valyl-l-prolyl-l-proline (vpp) and l-isoleucyl-l-prolyl-l-proline (ipp), when administered once daily for 91 consecutive days to rats. the test article, powdered casein hydrolysate (ch) known to contain 0.6% vpp plus ipp, was prepared using aspergillus oryzae protease. prior to administration to the rats by oral gavage, the test article was suspended in sterile water. groups of 12 m ...200516419578
studies of the toxicological potential of tripeptides (l-valyl-l-prolyl-l-proline and l-isoleucyl-l-prolyl-l-proline): vii. micronucleus test of tripeptides-containing casein hydrolysate and lactobacillus helveticus-fermented milk powders in rats and mice.the objective of these in vivo experiments was to assess the mutagenic potential of tripeptides as reflected by the ability of the test compound to induce the formation of micronuclei in mouse polychromatic erythrocytes. the test agents used in these experiments were (1) powdered aspergillus oryzae protease casein hydrolysate (ch) and (2) powdered lactobacillus helveticus-fermented milk (fm). both test agents contain two tripeptides, l-valyl-l-prolyl-l-proline (vpp) and l-isoleucyl-l-prolyl-l-pr ...200516419580
identification and analysis of ku70 and ku80 homologs in the koji molds aspergillus sojae and aspergillus oryzae.ku genes play a key role in the non-homologous end-joining pathway. we have identified ku70 and ku80 homologs in the koji molds aspergillus sojae and aspergillus oryzae, and have constructed the disruption mutants of ku70, ku80, and ku70-80 to characterize the phenotypic change in these mutants. neither ku70- nor ku80-disrupted strains show hypersensitivity to the dna damaging agents methylmethane sulfonate (mms) and phleomycin. moreover, undesirable phenotypes, such as poor growth or repressed ...200616428831
cladal relatedness among aspergillus oryzae isolates and aspergillus flavus s and l morphotype isolates.aspergillus flavus is the main etiological agent for aflatoxin contamination of crops. its close relative, a. oryzae, does not produce aflatoxins and has been widely used to produce fermented foods. we compared the phylogeny of a. oryzae isolates and l- and s-type sclerotial isolates of a. flavus using single nucleotide polymorphisms in the omta gene in the aflatoxin biosynthesis gene cluster and deletions in and distal to the norb-cypa intergenic region as phylogenetic signals. aflatoxin-produc ...200616430983
extraction of milk-clotting enzyme produced by solid state fermentation of aspergillus oryzae.studies on the extraction of milk-clotting enzyme after solid-state fermentation (ssf) of wheat bran by a local strain of aspergillus oryzae ls1 were done. the extraction of the enzyme was found to be depended on different parameters like nature of extractant, soaking time, temperature etc. from different inorganic and organic extractants, calcium chloride (0.05%) and glycerol (40%) were found to be the best solvents for leaching out milk-clotting enzyme. the optimum volume of calcium chloride w ...200516450841
growth and amylase production by aspergillus oryzae during solid state fermentation using banana waste as substrate.aspergillus oryzae, isolated from the starch rich litter soil, produced amylase when banana fruit stalk was used as substrate in a solid state fermentation system. the effects of incubation period, ph, temperature and various carbon sources on the production of amylase were studied. a maximum yield of 380u/mg was recorded on 96th hour of incubation. the amylase is tolerant to wide range of initial culture ph values (3 to 8) and temperature (25 to 35 degrees c), with an optimum ph of 5 and temper ...200516459551
direct assay for alpha-amylase using fluorophore-modified cyclodextrins.a simple assay method for alpha-amylase was developed based on fluorophore-modified cyclodextrins (cds). four kinds of cd derivatives bearing a 4-amino-7-nitrobenz-2-oxa-1,3-diazole (nbd-amine) moiety were prepared as artificial substrates for the assay method. the fluorescence intensity of all the nbd-amine-modified cds decreased upon addition of aspergillus oryzae alpha-amylase, indicating a reduction in hydrophobicity near the nbd-amine moiety induced by hydrolysis of the cd ring. nc4gammacd, ...200616464599
chemoprevention of mouse urinary bladder carcinogenesis by fermented brown rice and rice bran.fermented brown rice by aspergillus oryzae (fbra) has been shown to be a potent anti-carcinogenic compound. here, we investigated the modifying effects of dietary feeding with a naturally occurring anti-oxidant fbra on n-butyl-n-(4-hydroxybutyl)-nitrosamine (oh-bbn)-induced urinary bladder carcinogenesis in male icr mice. five-week-old male icr mice were divided into 7 groups, and groups 1-5 were given oh-bbn (500 ppm) in drinking water for 6 weeks starting at 7 weeks of age. groups 2 and 3 were ...200616465408
vacuolar membrane dynamics in the filamentous fungus aspergillus oryzae.vacuoles in filamentous fungi are highly pleomorphic and some of them, e.g., tubular vacuoles, are implicated in intra- and intercellular transport. in this report, we isolated aovam3, the homologue of the saccharomyces cerevisiae vam3 gene that encodes the vacuolar syntaxin, from aspergillus oryzae. in yeast complementation analyses, the expression of aovam3 restored the phenotypes of both deltavam3 and deltapep12 mutants, suggesting that aovam3p is likely the vacuolar and/or endosomal syntaxin ...200616467481
enhanced gene targeting frequency in ku70 and ku80 disruption mutants of aspergillus sojae and aspergillus the koji molds aspergillus sojae and aspergillus oryzae, exogenous dna is integrated in the genome, in most cases irrespective of the sequence homology, suggesting that dna integration occurs predominantly through a nonhomologous end joining pathway where two ku genes, namely, ku70 and ku80, play a key role. to determine the effect of ku gene disruption on the gene targeting frequency, we constructed ku70-, ku80-, and ku70-ku80-disrupted strains of a. sojae and a. oryzae. the gene targeting f ...200616470383
distribution profiles of isoflavone isomers in black bean kojis prepared with various filamentous fungi.this study was conducted to compare the transformation of both isoflavone derivatives (aglycones, beta-glucosides, and acetyl and malonyl gluclucosides) and beta-glucosidase activity in kojis fermented with various generally recognized as safe filamentous fungi including aspergillus awamori, aspergillus oryzae, aspergillus sojae, rhizopus azygosporus, and rhizopus sp. no. 2. solid fermentation was performed to prepare the kojis by inoculating the steamed black beans with starter organism and cul ...200616478253
effects of dietary oyster extract on lipid metabolism, blood pressure, and blood glucose in sd rats, hypertensive rats, and diabetic rats.oyster extract was prepared by hydrolysis of oyster protein with proteases, aloase (a protease from bacillus subtilis), and pancitase (a protease from aspergillus oryzae). rats were fed a diet containing 20% casein (the control diet) or 15% casein and 5% oyster extract (the oyster extract diet) as the protein source. the oyster extract diet exerted a significant reduction in serum cholesterol and liver triglyceride concentrations as compared with the control diet in sprague-dawley (sd) rats fed ...200616495664
cloning and expression of 1,2-alpha-mannosidase gene (fmanib) from filamentous fungus aspergillus oryzae: in vivo visualization of the fmanibp-gfp fusion protein.1,2-alpha-mannosidase catalyzes the specific cleavage of 1,2-alpha-mannose residues from protein-linked n-glycan. in this study, a novel dna sequence homologous to the authentic 1,2-alpha-mannosidase was cloned from a cdna library prepared from solid-state cultured aspergillus oryzae. the fmanib cdna consisted of 1530 nucleotides and encoded a protein of 510 amino acids in which all consensus motifs of the class i alpha-mannosidase were conserved. expression of the full length of 1,2-alpha-manno ...200616495665
[cloning and characterization of glyceraldehyde-3-phosphate dehydrogenase gene from chaetomium globosum].the amino acid sequence of glyceraldehyde 3-phosphate dehydrogenase (gapdh) gene from neurospora crassa (xp-327967) and colletotrichum lindemuthianu (p35143) were subjected to local tblastn searching against the ests local datebase of chaetomium globosum. the full length cdna sequence of 1240bp encoding gapdh gene with an open reading frame of 1014bp and encoding 337 amino acids was obtained. the protein molecular weight was 36.1kd. the dna sequence of gapdh gene was obtained through pcr amplifi ...200516496697
genomics of aspergillus fumigatus.aspergillus fumigatus is a filamentous fungal saprophyte that is ubiquitous in the environment. it is also a human pathogen and induces allergenic response, negatively impacting health care and associated costs significantly around the world. much of the basic biology of this organism is only poorly understood, but the recent completion and publication of its genome sequence provides an excellent tool for researchers to gain insight into these processes. in this review we will summarize some of ...200516499415
succession of bacterial and fungal communities during a traditional pot fermentation of rice vinegar assessed by pcr-mediated denaturing gradient gel electrophoresis.denaturing gradient gel electrophoresis (dgge) based on small subunit rrna gene was applied to a traditional rice vinegar fermentation process in which the conversion of rice starch into acetic acid proceeded in a pot. the fungal dgge profile indicated that the transition from aspergillus oryzae to saccharomyces sp. took place at the initial stage at which alcohol production was observed. the early stage was characterized by the coexistence of saccharomyces sp. and lactic acid bacteria. almost a ...200616499984
characterization of a periplasmic s1-like nuclease coded by the mesorhizobium loti symbiosis island.dna sequences encoding hypothetical proteins homologous to s1 nuclease from aspergillus oryzae are found in many organisms including fungi, plants, pathogenic bacteria, and eukaryotic parasites. one of these is the m1 nuclease of mesorhizobium loti which we demonstrate herein to be an enzymatically active, soluble, and stable s1 homolog that lacks the extensive mannosyl-glycosylation found in eukaryotic s1 nuclease homologs. we have expressed the cloned m1 protein in m. loti and purified recombi ...200616529713
evidence of rip (repeat-induced point mutation) in transposase sequences of aspergillus oryzae.a dna methyl-binding column was used to isolate genomic fragments enriched for dna-methylation from aspergillus parasiticus. one of the isolated sequences presented 67% identity at the protein level with the transposase from the transposable element tan1 of aspergillus niger var. awamori, and was found to be present in at least 20 copies in the aspergillus oryzae database. analysis of four copies showed evidence of c:g to t:a transitions in at least 98.2% of the mutations found over a 1,032-1,18 ...200616531081
combined expression of aspergillus nidulans endoxylanase x24 and aspergillus oryzae (alpha)-amylase in industrial baker's yeasts and their use in bread making.the aspergillus nidulans endoxylanase x24 and the aspergillus oryzae (alpha)-amylase cdnas were placed under the control of the saccharomyces cerevisiae actin promoter (pact1) and introduced into baker's yeast. bread made with transformants expressing both enzymes (yepact-amy-act-x24) showed a 30% increase in volume and reduced firmness in comparison with that produced with a commercial strain. endoxylanase x24 and (alpha)-amylase seem to act synergistically to improve the quality of bread in te ...199616535419
an enzymatically produced novel cyclomaltopentaose cyclized from amylose by an alpha-(1-->6)-linkage, cyclo-{-->6)-alpha-d-glcp-(1-->4)-alpha-d-glcp-(1-->4)-alpha-d-glcp-(1-->4)-alpha-d-glcp-(1-->4)-alpha-d-glcp-(1-->}.a bacterial strain am7, isolated from soil and identified as bacillus circulans, produced two kinds of novel cyclic oligosaccharides. the cyclic oligosaccharides were produced from amylose using a culture supernatant of the strain as the enzyme preparation. the major product was a cyclomaltopentaose cyclized by an alpha-(1-->6)-linkage, cyclo-{-->6)-alpha-d-glcp-(1-->4)-alpha-d-glcp-(1-->4)-alpha-d-glcp-(1-->4)-alpha-d-glcp-(1-->4)-alpha-d-glcp-(1-->}. the other minor product was cyclomaltohexao ...200616545346
g-protein and camp-mediated signaling in aspergilli: a genomic perspective.we have carried out an in silico exploration of the genomes of aspergillus nidulans, aspergillus fumigatus, and aspergillus oryzae, and identified components of g-protein/camp-mediated signaling. putative g-protein coupled receptors (gpcrs) were distributed over nine classes. the gpcrs within classes were well conserved among aspergilli but varied in other ascomycetes. as previously observed in a. nidulans and other fungi, three galpha, one gbeta, and one ggamma subunits of g proteins were ident ...200616546420
deletion analysis of the superoxide dismutase (sodm) promoter from aspergillus oryzae.the manganese superoxide dismutase gene (sodm) is very highly expressed in aspergillus oryzae. to elucidate the basis for this high-level expression, deletion analysis of the promoter was undertaken using beta-glucuronidase (gus) as a reporter. deletion of a 63-bp sequence from -200 to -138 in the 1,038-bp sodm promoter caused a drastic decrease in gus activity. in addition, an electrophoretic gel mobility shift assay (emsa) implicated a 30-bp element from -209 to -178 containing cis-element(s) ...200616547700
suppression on the mutagenicity of 4-nitroquinoline-n-oxide by the methanol extracts of soybean koji prepared with various filamentous this study, solid fermentation of soybean with various gras filamentous fungi including aspergillus sojae bcrc 30103, aspergillus oryzae bcrc 30222, aspergillus awamori, actinomucor taiwanesis and rhizopus sp. was performed to prepare various soybean kojis. toxicity, mutagenicity and suppression on the mutagenesis induced by a direct mutagen, 4-nitroquinoline-n-oxide (4-nqo) on salmonella typhimurium ta 100, by the various methanol extracts of the prepared soybean koji and unfermented soybean ...200616580082
novel hydrophobic surface binding protein, hsba, produced by aspergillus oryzae.hydrophobic surface binding protein a (hsba) is a secreted protein (14.5 kda) isolated from the culture broth of aspergillus oryzae rib40 grown in a medium containing polybutylene succinate-co-adipate (pbsa) as a sole carbon source. we purified hsba from the culture broth and determined its n-terminal amino acid sequence. we found a dna sequence encoding a protein whose n terminus matched that of purified hsba in the a. ozyzae genomic sequence. we cloned the hsba genomic dna and cdna from a. ory ...200616597938
enhancement of enantioselectivity in the bacillus subtilis protease-catalyzed hydrolysis of n-free amino acid esters using the ester grouping-modification approach.the generality of enantioselectivity enhancement through the modification of the alcohol moiety of a substrate ester was ascertained, for in the bacillus subtilis protease-catalyzed hydrolysis of n-unprotected amino acid esters the enantioselectivity was enhanced largely by switching the conventional methyl ester to esters with a longer alkyl chain such as the isobutyl ester (from e = 3 to e = 130-170 in the case of 4-fluorophenylalanine esters) as in the enzymatic hydrolysis mediated by aspergi ...200616614915
the region in a subunit of the aspergillus ccaat-binding protein similar to the hap4p-recruiting domain of saccharomyces cerevisiae hap5p is not essential for transcriptional enhancement.the ccaat-binding complex in aspergillus species, known as the hap complex, consists of at least three subunits, hapb, hapc, and hape. each hap subunit contains an evolutionarily conserved core domain. in this study, a series of the truncated gene, which encodes the hape subunit of aspergillus oryzae, was constructed to survey the regions essential for the transcriptional enhancement of fungal genes. it was revealed that the non-conserved regions and the conserved region similar to the hap4p rec ...200616636442
lentinula edodes tlg1 encodes a thaumatin-like protein that is involved in lentinan degradation and fruiting body senescence.lentinan is an antitumor product that is purified from fresh lentinula edodes fruiting bodies. it is a cell wall component, comprising beta-1,3-glucan with beta-1,6-linked branches, which becomes degraded during postharvest preservation as a result of increased glucanase activity. in this study, we used n-terminal amino acid sequence to isolate tlg1, a gene encoding a thaumatin-like (tl) protein in l. edodes. the cdna clone was approximately 1.0 kb whereas the genomic sequence was 2.1 kb, and co ...200616648221
substrate specificity of aspergillus oryzae family 3 beta-glucosidase.among glycoside hydrolases, beta-glucosidase plays a unique role in many physiological and biocatalytical processes that involve the beta-linked o-glycosyl bond of various oligomeric saccharides or glycosides. structurally, the enzyme can be grouped into glycoside hydrolase family 1 and 3. although the basic ("retaining, double-displacement") mechanism for the catalysis of family 3 beta-glucosidase has been established, in-depth understanding of its structure-function relationship, particularly ...200616650812
proteomic analysis of extracellular proteins from aspergillus oryzae grown under submerged and solid-state culture conditions.filamentous fungi are widely used for the production of homologous and heterologous proteins. recently, there has been increasing interest in aspergillus oryzae because of its ability to produce heterologous proteins in solid-state culture. to provide an overview of protein secretion by a. oryzae in solid-state culture, we carried out a comparative proteome analysis of extracellular proteins in solid-state and submerged (liquid) cultures. extracellular proteins prepared from both cultures sequen ...200616672490
extracellular production of neoculin, a sweet-tasting heterodimeric protein with taste-modifying activity, by aspergillus oryzae.neoculin (ncl), a protein with sweetness approximately 500-fold that of sugar, can be utilized as a nonglycemic sweetener. it also has taste-modifying activity to convert sourness to sweetness. ncl is a heterodimer composed of an n-glycosylated acidic subunit (nas) and a basic subunit (nbs), which are conjugated by disulfide bonds. for the production of recombinant ncl (rncl) by aspergillus oryzae, alpha-amylase with a kex2 cleavage site, -k-r-, was fused upstream of each of nas and nbs and the ...200616672522
aspergillus oryzae strains with a large deletion of the aflatoxin biosynthetic homologous gene cluster differentiated by chromosomal breakage.recently we divided aspergillus oryzae rib strains into group 1, having seven aflatoxin biosynthesis homologous genes (aflt, nor-1, aflr, nora, avna, verb, and vbs), and group 2, having three homologues (avna, verb, and vbs). here, partial aflatoxin homologous gene cluster of rib62 from group 2 was sequenced and compared with that of rib40 from group 1. rib62 showed a large deletion upstream of ver-1 with more than half of the aflatoxin homologous gene cluster missing including aflr, a positive ...200616673111
[production of antitumor substances from shochu distillation remnants].this study aimed to develop a new processing method for the effective use of rice shochu distillation remnants. we examined the inhibitory effects on the growth of human lung carcinoma cells in the medium of rice shochu distillation remnants with various fungi. interestingly, high inhibitory effects on the growth of human lung carcinoma cells in the medium of rice shochu distillation remnants with aspergillus oryzae were obtained, although no inhibitory effect was observed in the case of synthet ...200616679744
[genome sequencing of aspergillus oryzae]. 200616686348
development of aflp-derived, functionally specific markers for environmental persistence studies of fungal strains.the ability to rapidly identify and quantify a microbial strain in a complex environmental sample has widespread applications in ecology, epidemiology, and industry. in this study, we describe a rapid method to obtain functionally specific genetic markers that can be used in conjunction with standard or real-time polymerase chain reaction (pcr) to determine the abundance of target fungal strains in selected environmental samples. the method involves sequencing of randomly cloned aflp (amplified ...200616699570
square-plate culture method allows detection of differential gene expression and screening of novel, region-specific genes in aspergillus oryzae.when grown on solid agar medium, the mycelium of a filamentous fungus, aspergillus oryzae, forms three morphologically distinct regions: the tip (t), white (w), and basal (b) regions. in this study, we developed the square-plate culture method, a novel culture method that enabled the extraction of mrna samples from the three regions and analyzed the differential gene expression of the a. oryzae mycelium in concert with the microarray technique. expression of genes involved in protein synthesis w ...200616708193
functional expression of candida antarctica lipase b in eschericha coli.candida antarctica lipase b (calb) is an important catalyst in bio-organic synthesis. to optimize its performance, either the reaction medium is changed or the lipase itself is modified. in the latter case, mutants are generated in eschericha coli and subsequently expressed in fungal hosts for their characterization. here we present the functional expression of calb in the periplasm of e. coli. by step-wise deletion of the calb signal and propeptide we were able to express and purify two differe ...200616713003
alpha-galactosidase production by aspergillus oryzae in solid-state fermentation.comparisons were made for alpha-galactosidase production using red gram plant waste (rgpw) with wheat bran (wb) and other locally available substrates using the fungus aspergillus oryzae under solid-state fermentation (ssf). rgpw proved to be potential substrate for alpha-galactosidase production as it gave higher enzyme titers (3.4 u/g) compared to wb (2.7 u/g) and other substrates tested. mixing wb with rgpw (1:1, w/w) resulted enhanced alpha-galactosidase yield. the volume of moistening agent ...200716713256
improvement of the glab promoter expressed in solid-state fermentation (ssf) of aspergillus oryzae.the glucoamylase-encoding gene (glab) promoter should be very useful for recombinant protein production in solid-state fermentation (ssf) of aspergillus oryzae. a 97-bp fragment containing the cis-element of the glab promoter was inserted into the glaa promoter, which was little expressed in ssf. the chimeric promoter showed about a 24-fold increase in promoter activity in ssf. eight copies of the 97-bp fragment were tandemly fused with the glab promoter. the improved promoter showed about a 4.6 ...200616717420
differential distribution of the endoplasmic reticulum network as visualized by the bipa-egfp fusion protein in hyphal compartments across the septum of the filamentous fungus, aspergillus oryzae.we visualized the endoplasmic reticulum (er) network by expression of the bipa-egfp fusion protein in the filamentous fungus, aspergillus oryzae, and focused on the spatial difference of the er distribution throughout hyphae. the er formed an interconnected network with motility and displayed a gradient distribution from the apical region. the er was also found as a tubular network along the septum, which was formed soon after the completion of septation. discontinuity of the er network distribu ...200616759887
food-processing enzymes from recombinant microorganisms--a review.enzymes are commonly used in food processing and in the production of food ingredients. enzymes traditionally isolated from culturable microorganisms, plants, and mammalian tissues are often not well-adapted to the conditions used in modern food production methods. the use of recombinant dna technology has made it possible to manufacture novel enzymes suitable for specific food-processing conditions. such enzymes may be discovered by screening microorganisms sampled from diverse environments or ...200616769167
reduction of flatus-inducing factors in soymilk by immobilized alpha-galactosidase.alpha-galactosidase from aspergillus oryzae was immobilized on chitosan beads using glutaraldehyde as a cross-linking agent. the general properties of free and immobilized enzymes were determined. the optimum ph for the free and immobilized enzymes was 4.8 and 4.6 respectively. the optimum temperature for the free enzyme was 50 degrees c, whereas that of immobilized enzyme was increased to 56 degrees c. kinetic parameters were determined with synthetic substrate (p-nitrophenyl alpha-d-galactopyr ...200616780418
production of high-content galacto-oligosaccharide by enzyme catalysis and fermentation with kluyveromyces marxianus.of three beta-galactosidases from aspergillus oryzae, kluyveromyces lactis and bacillus sp., used for the production of low-content galacto- oligosaccharides (gos) from lactose, the latter produced the highest yield of trisaccharides and tetrasaccharides. gos production was enhanced by mixing beta-galactosidase glucose oxidase. the low-content gos syrups, produced either by beta-galactosidase alone or by the mixed enzyme system, were subjected to the fermentation by kluyveromyces marxianus, wher ...200616786243
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