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enzymatic synthesis of 2-acetamido-4-o-(2-acetamido-2-deoxy-beta-d- galactopyranosyl)-2-deoxy-d-glucopyranose and 2-acetamido-6-o-(2-acetamido-2- deoxy-beta-d-galactopyranosyl)-2-deoxy-d-glucopyranose catalysed by the beta-n-acetylhexosaminidase from aspergillus oryzae. 19958593628
cloning, characterization and overproduction of nuclease s1 gene (nucs) from aspergillus oryzae.the nuclease s1 gene (nucs) from aspergillus oryzae was isolated using a polymerase-chain-reaction-amplified dna fragment as a probe, and a 2.6-kb sali-ecori fragment containing the nucs gene was sequenced. it was deduced that the nucs gene had two short introns, 49 and 50 nucleotides in length. the nucs gene had an open-reading frame of 963 base pairs and coded for a protein of 287 amino acid residues, comprising the signal peptide of 20 amino acids and a mature protein of 267 amino acids. the ...19958597544
influence of protein conformation on disulfide bond formation in the oxidative folding of ribonuclease t1.in oxidative protein folding the interdependence between the acquisition of an ordered native-like conformation and disulfide bond formation was investigated by using the c2s/c10n variant of ribonuclease t1 as a model. this protein of 104 residues has a single disulfide bond between cys6 and cys103. in the reduced form it is unfolded in the presence of urea, but native-like folded when > or = 1.5 m nacl is present. the influence of a folded conformation on the individual thiol/disulfide exchange ...19957643382
effects of lactitol-oligosaccharides on the intestinal microflora in rats.lactitol-oligosaccharide (lo) was prepared from lactitol by a transgalactosylation reaction catalyzed by aspergillus oryzae beta-galactosidase. the utilization of lo by human intestinal bacteria, the digestion of lo by rat jejunum mucosal homogenates and the effects of lo on the intestinal microflora in rats were compared with those of lactitol. 1) lo was utilized in vitro by bifidobacterium, but lactitol was not utilized. 2) neither lo nor lactitol were digested by rat jejunum mucosal homogenat ...19957616329
molecular cloning and characterization of a rhamnogalacturonan acetylesterase from aspergillus aculeatus. synergism between rhamnogalacturonan degrading enzymes.a rhamnogalacturonan acetylesterase (rgae) was purified to homogeneity from the filamentous fungus aspergillus aculeatus, and the nh2-terminal amino acid sequence was determined. full-length cdnas encoding the enzyme were isolated from an a. aculeatus cdna library using a polymerase chain reaction-generated product as a probe. the 936-base pair rha1 cdna encodes a 250-residue precursor protein of 26,350 da, including a 17-amino acid signal peptide. the rha1 cdna was overexpressed in aspergillus ...19957592973
purification and characterization of a novel specific phosphatidylglycerol-phosphatidylinositol transfer protein with high activity from aspergillus oryzae.a novel phospholipid transfer protein has been purified to homogeneity 406-fold from the filamentous fungus aspergillus oryzae. the successive steps of purification comprised ultrafiltration, gel filtration on sephadex g-75, ion exchange chromatographies on deae-sepharose and mono q. the active protein is a monomer with a molecular mass of 19,000, estimated from sds electrophoresis, amino acid composition as well as gel filtration. the isoelectric point is 4.8. the amino acid composition is char ...19957742351
isolation and characterization of polygalacturonase genes (peca and pecb) from aspergillus flavus.two genes, peca and pecb, encoding endopolyglacturonases were cloned from a highly aggressive strain of aspergillus flavus. the peca gene consisted of 1,228 bp encoding a protein of 363 amino acids with a predicted molecular mass of 37.6 kda, interrupted by two introns of 58 and 81 bp in length. accumulation of peca mrna in both pectin- or glucose-grown mycelia in the highly aggressive strain matched the activity profile of a pectinase previously identified as p2c. transformants of a weakly aggr ...19957574642
the nitrate reductase gene from a shoyu koji mold, aspergillus oryzae kbn616.a niad gene encoding nitrate reductase was isolated from aspergillus oryzae kbn616 and sequenced. the structural gene comprises 2973 bp and 868 amino acids, which showed a high degree of similarity to nitrate reductases from other filamentous fungi. the coding sequence is interrupted by six introns varying in size from 48 to 98 bp. the intron positions are all conserved among the niad genes from a. oryzae, aspergillus nidulans, and aspergillus niger. a homologous transformation system was develo ...19958520125
cloning, sequence analysis and expression in yeasts of a cdna containing a lipomyces kononenkoae alpha-amylase-encoding gene.the yeast lipomyces kononenkoae (lk) secretes a highly active raw starch-degrading alpha-amylase (alpha amy) that liberates reducing groups from glucose polymers containing both alpha-1,4 and alpha-1,6 bonds. the lka1 gene encoding this industrially important alpha amy was cloned as a 2261-bp cdna fragment from a glucose-derepressed mutant (igc4052b) of lk and characterized. the nucleotide (nt) sequence of the cdna fragment was determined, revealing an open reading frame of 1872 bp, encoding a 5 ...19958529895
cloning and nucleotide sequence of the ribonuclease t1 gene (rnta) from aspergillus oryzae and its expression in saccharomyces cerevisiae and aspergillus oryzae.a genomic dna encoding ribonuclease (rnase) t1 from aspergillus oryzae was cloned using a synthetic oligonucleotide probe. the cloned gene (designated rnta) encoded functional rnase t1, since an a. oryzae transformant with multiple copies of the rnta gene showed higher rnase t1 activity (over 200 times) than a transformant with a vector. a cdna was cloned by reverse transcription polymerase chain reaction (rt-pcr) with primers corresponding to the 5' terminus and 3' terminus of the reading frame ...19958534978
enzymatic synthesis of peptides containing unnatural amino acids.several proteases were studied as potential catalysts for the enzymatic synthesis of oligopeptides containing the unnatural amino acid allylglycine, the overall objective being the synthesis of a reactive tetrapeptide that could be chemically polymerized into a potentially biocompatible or biodegradable material. commercially available enzymes were screened for esterase activity toward the methyl ester of the amino acid allylglycine (dl-agome) to identify potential catalysts for dipeptide synthe ...19958541021
close evolutionary relatedness among functionally distantly related members of the (alpha/beta)8-barrel glycosyl hydrolases suggested by the similarity of their fifth conserved sequence region.a short conserved sequence equivalent to the fifth conserved sequence region of alpha-amylases (173_lpdld, aspergillus oryzae alpha-amylase) comprising the calcium-ligand aspartate, asp-175, was identified in the amino acid sequences of several members of the family of (alpha/beta)8-barrel glycosyl hydrolases. despite the fact that the aspartate is not invariantly conserved, the stretch can be easily recognised in all sequences to be positioned 26-28 amino acid residues in front of the well-know ...19958543020
molecular cloning and nucleotide sequence of the protyrosinase gene, melo, from aspergillus oryzae and expression of the gene in yeast cells.the coding region of the protyrosinase gene, melo, from aspergillus oryzae occupies 1671 base pairs of the genomic dna and is separated into two exons by one intron. the full-length cdna of the melo gene was cloned. analysis of the 1617 base pairs nucleotide sequence revealed a single open reading frame coding 539 amino acid residues. the cdna has been expressed in yeast cells. the predicted protein product derived from the melo gene is identified by western blotting and activity determination. ...19957893753
modulation of the behavior of a protein in polyacrylamide gel electrophoresis in the presence of dodecyl sulfate by varying the cations.the denaturation of aspergillus oryzae alpha-amylase by dodecyl sulfates can be modulated by the change of cations among sodium, lithium, and several alkanolammonium ions. the denaturation can be detected by polyacrylamide gel electrophoresis in the presence of alkanolammonium dodecyl sulfates, particularly around 0 degree c where the denaturation reaction can be virtually frozen. the moderate stability of the amylase against denaturation by dodecyl sulfates helped to demonstrate the difference ...19957733460
reduction of cyanide levels in cassava during sequential sundrying and solid state fermentation.the cyanide levels were followed during protein enrichment of cassava by the fungus aspergilus oryzae. the total cyanogen level decreased by 158 mg/kg dry weight to 54.2 mg/kg dry weight as a result of the whole process including fermentation. the cyanogenic glucoside level decreased by 88% during the fermentation process while acetone cyanohydrin was retained in the cassava. the prefermentation processing which involved crushing, sundrying and milling the cassava into flour, reduced the total c ...19957712337
induction and repression of alpha-amylase production in batch and continuous cultures of aspergillus oryzae.the intra- and extracellular concentrations of alpha-amylase in aspergillus oryzae have been measured during batch culture of a wild-type strain and two recombinant strains. the mean intracellular level for the two recombinant strains was about four to five times the level of the wild-type strain. the recombinant strains also had a higher alpha-amylase productivity, whereas the residence time of the intracellular alpha-amylase pool was approximately the same for the three strains. at high glucos ...19957582005
sequence variability in homologs of the aflatoxin pathway gene aflr distinguishes species in aspergillus section flavi.the aspergillus parasiticus aflr gene, a gene that may be involved in the regulation of aflatoxin biosynthesis, encodes a putative zinc finger dna-binding protein. pcr and sequencing were used to examine the presence of aflr homologs in other members of aspergillus section flavi. the predicted amino acid sequences indicated that the same zinc finger domain, ctscasskvrctkekpacarcierglac, was present in all of the aspergillus sojae, aspergillus flavus, and aspergillus parasiticus isolates examined ...19957887625
cloning and nucleotide sequence of the calmodulin-encoding gene (cmda) from aspergillus oryzae.a cdna and genomic gene encoding calmodulin were isolated from aspergillus oryzae using a part of the calmodulin gene from a. nidulans as a hybridization probe. the gene was in a 3.4-kb sphi fragment and southern-blot analysis of genomic dna suggested the existence of a single copy of the calmodulin gene in a. oryzae. the nucleotide sequence analysis showed that the gene consists of five introns and six exons. although the nucleotide sequence homology with that of a. nidulans was not so high (68 ...19957549095
nucleotide sequence and expression of alpha-glucosidase-encoding gene (agda) from aspergillus oryzae.we have isolated an alpha-glucosidase(agl)-encoding gene (agda) from aspergillus oryzae by heterologous hybridization using the corresponding aspergillus niger gene as a probe. southern hybridization analysis showed that the agda gene is on a 5.0-kb scai fragment and there is a single copy in the a. oryzae chromosome. comparison with the a. niger agda gene indicated that the agda gene contains three putative introns from 52 to 59 nucleotides long, and that it encodes 985 amino acid residues. the ...19957549103
gene transfer by electroporation of filamentous fungi. 19957550746
molecular cloning of the cdna and gene for an elastinolytic aspartic proteinase from aspergillus fumigatus and evidence of its secretion by the fungus during invasion of the host lung.hydrolysis of structural proteins in the lung by extracellular proteinases secreted by aspergillus fumigatus is thought to play a significant role in invasive aspergillosis. this fungus was found previously to secrete an elastinolytic serine proteinase and a metalloproteinase. we report that a. fumigatus also secretes an aspartic proteinase (aspergillopepsin f) that can catalyze hydrolysis of the major structural proteins of basement membrane, elastin, collagen, and laminin. the ph optimum for t ...19957558282
an acetylglucomannan esterase of aspergillus oryzae; purification, characterization and role in the hydrolysis of o-acetyl-galactoglucomannan.an acetyl glucomannan esterase (agme) was purified to electrophoretic homogeneity from the culture supernatant of aspergillus oryzae. this new enzyme had a molecular mass of 36 kda and an isoelectric point of 4.6. it was most active in the ph range 5.0-5.5 and was stable for 24 h at 40 degrees c at ph 5.0-6.0. the purified esterase liberated acetic acid from o-acetyl-galactoglucomannan, o-acetyl-4-o- methylglucuronoxylan and alpha-naphtyl acetate. the specific activity was 10-times higher for ac ...19957576539
skin-prick tests for hypersensitivity to alpha-amylase preparations.twenty-five asthmatic subjects with suspected alpha-amylase hypersensitivity were studied by skin-prick tests, a capture elisa, immunoblotting and bronchial provocation tests. at the same time, different amylases were analysed by sds-page and immunoblotting using a polyclonal rabbit antiserum. eight patients showed a positive bronchial response to amylase. seven of them had positive skin-prick tests, with this method being the most sensitive approach for diagnosis. however, in four cases, skin t ...19957605978
isolation and characterization of the gene encoding the surface membrane 3'-nucleotidase/nuclease of leishmania donovani.leishmania donovani and related trypanosomatid protozoa possess an externally oriented surface membrane enzyme capable of hydrolyzing both 3'-nucleotides and nucleic acids. by virtue of these activities, this 3'-nucleotidase/nuclease (3'-nt/nu), previously shown to be analogous to fungal and plant class-i single-strand-specific nucleases, is thought to play a critical role in the salvage of purines, essential for the survival of these organisms. the 43-kda 3'-nt/nu was purified from l. donovani ...19957630383
purification and properties of alpha-amylase from aspergillus oryzae atcc 76080.an alpha-amylase was purified from the solid cultural extract of aspergillus oryzae atcc 76080 by sequential steps of amylopectin affinity adsorption, deae-sepharose ion-exchange chromatography and sephacryl s-200 hr gel filtration. by these steps, the purity of the enzyme increased by 16 fold and recovery of the enzyme activity was 45%. the purified enzyme had an optimal ph between 4 to 5, optimal temperature at 50 degrees c and a km value of 0.22% for hydrolysis of starch. about 80% of the enz ...19957663414
purification and characterization of a lipase from aspergillus oryzae.a lipase from aspergillus oryzae was purified by ammonium sulfate fractionation, anion exchange chromatography, hydrophobic interaction chromatography, and anion exchange chromatography. the purified enzyme was a monomeric protein with a molecular mass of 41 kda estimated by sds-page and 39 kda by gel filtration. the optimum ph at 30 degrees c and optimum temperature at ph 7.0 were 7.0 and 30 degrees c, respectively. the enzyme was stable over a ph range of 6-9 at 25 degrees c for 18 h, and up t ...19957670177
genome structure and nucleotide sequence of a lipolytic enzyme gene of aspergillus oryzae.aspergillus oryzae, which is widely used for japanese traditional fermentation, produced at least two lipolytic enzymes (l1 and l2). southern hybridization analysis of restriction enzyme-digested genomic dna fragments of aspergillus oryzae with 23-mer oligonucleotides synthesized according to the amino acid sequence of the enzyme l1 as probes suggested that there is single copy of the l1 gene in the genome. dna fragments containing the l1 gene were cloned in escherichia coli. nucleotide sequenci ...19957705606
disulfide bonds and glycosylation in fungal peroxidases.four conserved disulfide bonds and n-linked and o-linked glycans of extracellular fungal peroxidases have been identified from studies of a lignin and a manganese peroxidase from trametes versicolor, and from coprinus cinereus peroxidase (cip) and recombinant c. cinereus peroxidase (rcip) expressed in aspergillus oryzae. the eight cysteine residues are linked 1-3, 2-7, 4-5 and 6-8, and are located differently from the four conserved disulfide bridges present in the homologous plant peroxidases. ...19957851395
alpha-amylase contained in bread can induce food allergy. 19957822654
biological activity of a heptaketide metabolite from pleiochaeta setosa.an uncommon heptaketide metabolite, setosol (2,8-dimethyl-4 methoxy-6,10,11-trihydroxy-benzo-oxaonin), was isolated from a liquid culture filtrate of the fungus pleiochaeta setosa. the biological activity of the molecule was studied by using 12 microbial strains consisting of three bacteria, three yeasts and six fungi. the level of activity was compared with those of known antibiotics and antifungal agents. the metabolite exhibited antifungal and antibiotic activity against drechslera oryzae, ge ...19957766015
industrial application of immobilized biocatalysts in japan. 19957785866
expression cloning, purification and characterization of a beta-1,4-galactanase from aspergillus aculeatus.expression cloning has been used to isolate a cdna encoding beta-1,4-galactanase from the filamentous fungus aspergillus aculeatus. a cdna library was prepared from mycelia, inserted in a yeast expression vector and transformed into saccharomyces cerevisiae. thirteen clones secreting galactanase activity were identified from a screening of approximately 2.5 x 10(4) yeast colonies. all clones expressed transcripts of the same galactanase gene. the cdna was re-cloned in an aspergillus expression v ...19957788716
stability of ribonuclease t2 from aspergillus oryzae.the stability of ribonuclease t2 (rnase t2) from aspergillus oryzae against guanidine hydrochloride and heat was studied by using cd and fluorescence. rnase t2 unfolded and refolded reversibly concomitant with activity, but the unfolding and refolding rates were very slow (order of hours). the free energy change for unfolding of rnase t2 in water was estimated to be 5.3 kcal.mol-1 at 25 degrees c by linear extrapolation method. from the thermal unfolding experiment in 20 mm sodium phosphate buff ...19957795525
a system for production of commercial quantities of human lactoferrin: a broad spectrum natural antibiotic.we previously reported the production of limited quantities of biologically active recombinant human lactoferrin in the filamentous fungus aspergillus oryzae. in the present study, we report a modification of this production system combined with a classical strain improvement program that has enabled production of levels of recombinant human lactoferrin in excess of 2 g/l. the protein was expressed in aspergillus awamori as a glucoamylase fusion polypeptide which was secreted into the growth med ...19959634791
molecular cloning and sequence analysis of a gene encoding an aspartic proteinase from aspergillus oryzae. 19958540376
variability in heat-induced fragmentation of a protein in the presence of dodecyl sulfate: the role of an intramolecular sulfhydryl/disulfide exchange.alpha-amylase from aspergillus oryzae was investigated to better understand how disulfide-bonded proteins behave during denaturation with dodecyl sulfate. it was previously reported that the alpha-amylase, when denatured with dodecyl sulfate, forms two species, d1 and d2. in d1, the disulfide bonds remain intact, while in d2, there is a restricted single sulfhydryl/disulfide (sh/ss) exchange reaction. this phenomenon was re-examined as follows: electrophoretic analysis of fragments created with ...19958720123
effect of probiotic supplementation on serum/yolk cholesterol and on egg shell thickness in layers.1. the effect of probiotic supplementation on egg production, on serum and yolk cholesterol and on egg shell thickness in 24 white leghorn layers was studied from 28-38 weeks of age. 2. in 3 treatments the diet was supplemented with 0, 100 and 150 mg probiotic/kg food. 3. in the 100 mg probiotic group, egg production improved by 5%, and shell thickness improved slightly, with fewer thin-shelled eggs than in the control (8.6% compared to 18.6%). 4. the initial serum cholesterol concentration of 1 ...19958746981
characteristic expression of three amylase-encoding genes, agda, amyb, and glaa in aspergillus oryzae transformants containing multiple copies of the agda gene.in aspergillus oryzae wild-type strains, the expression of the agda gene encoding alpha-glucosidase (agl) is induced by maltose at the transcriptional level in a similar manner to the amyb gene encoding takaamylase a (taa) and the glaa gene encoding glucoamylase (gla). in a. oryzae transformants containing multiple copies of the agda gene, a high-level of agl activity was observed. this was accompanied by a significant reduction in taa and gla activities. moreover, transformants with the highest ...19958611747
s1 nuclease: immunoaffinity purification and evidence for the proximity of cysteine 25 to the substrate binding site.a simple procedure, involving heat treatment, gel filtration on sephadex-g 100 followed by chromatography on anti-s1 nuclease antibodies bound to sepharose, was developed for purification of s1 nuclease to homogeneity with an overall yield of 72%. s1 nuclease was rapidly inactivated, at ph 6.0 and 37 degrees c, in presence of o-phthalaldehyde. kinetic analysis of o-phthalaldehyde medicated inactivation showed that the reaction followed pseudo-first-order kinetics and the loss of enzyme activity ...19958619949
functionally essential, invariant glutamate near the c-terminus of strand beta 5 in various (alpha/beta)8-barrel enzymes as a possible indicator of their evolutionary relatedness.twelve different (alpha/beta)8-barrel enzymes belonging to three structurally distinct families were found to contain, near the c-terminus of their strand beta 5, a conserved invariant glutamic acid residue that plays an important functional role in each of these enzymes. the search was based on the idea that a conserved sequence region of an (alpha/beta)8-barrel enzyme should be more or less conserved also in the equivalent part of the structure of the other enzymes with this folding motif owin ...19958637850
structural consequences of neopullulanase mutations.bacillus stearothermophilus neopullulanase (npl) structure was modeled based on aspergillus oryzae alpha-amylase (taa) to understand the structure-function relationships of this pullulan hydrolyzing enzyme. the npl structure seems to consist of a central (alpha/beta)8 barrel to which the other domains are attached. the immediate surroundings of the npl catalytic site were found to have very similar structure to taa. the more distant sites are different due to the stereochemical requirements of a ...19968695646
effects of saccharomyces cerevisiae and aspergillus oryzae cultures on ruminal fermentation in dairy cows.four lactating holstein cows, fitted with ruminal and duodenal cannulas, were used in a 4 x 4 latin square design to examine the effects of supplemental yeast (saccharomyces cerevisiae) and fungal (aspergillus oryzae) cultures on ruminal fermentation, microbial populations, and nutrient supply to the small intestine. cows were fed a basal diet comprising 32.5% corn silage, 17.5% alfalfa hay, 35.3% corn grain, 12.7% soybean meal, and 2% vitamin and mineral mixture on a dm basis. treatments were a ...19968708102
identification of the active-site peptide of 2,3-dihydroxybenzoic acid decarboxylase from aspergillus oryzae.the non-oxidative decarboxylation of aromatic acids is a poorly understood reaction. the transformation of 2,3-dihydroxybenzoic acid to catechol in the fungal metabolism of indole is a prototype of such a reaction. 2,3-dihydroxybenzoic acid decarboxylase (ec 4.1.1.46) which catalyzes this reaction was purified to homogeneity from anthranilate induced cultures of aspergillus oryzae using affinity chromatography. the enzyme did not require cofactors like nad+, plp, tpp or metal ions for its activi ...19968620029
invariant glycines and prolines flanking in loops the strand beta 2 of various (alpha/beta)8-barrel enzymes: a hidden homology?the question of parallel (alpha/beta)8-barrel fold evolution remains unclear, owing mainly to the lack of sequence homology throughout the amino acid sequences of (alpha/beta)8-barrel enzymes. the "classical" approaches used in the search for homologies among (alpha/beta)8-barrels (e.g., production of structurally based alignments) have yielded alignments perfect from the structural point of view, but the approaches have been unable to reveal the homologies. these are proposed to be "hidden" in ...19968762144
the effect of saccharomyces cerevisiae and aspergillus oryzae on fermentations in the rumen of faunated and defaunated sheep; protozoal and probiotic interactions.we measured the effect of the direct addition to the rumen of saccharomyces cerevisiae (sc 50 mg/day) and aspergillus oryzae (ao 3 g/day) on the fermentation processes in fistulated sheep. the measurements were carried out on animals whose rumens were first defaunated and then refaunated. the animals received a ration composed of hay (600 g/day), barley (600 g/day) and soybean meal (150 g/day), fed twice daily in two equal meals. the number of fungi and total, viable or cellulolytic bacteria wer ...19968766732
effect of probiotic supplementation on growth, nitrogen utilisation and serum cholesterol in broilers.1. the effect of dietary probiotic supplementation on the growth, nitrogen utilisation and serum cholesterol content of broiler chickens was studied in 2 trials. 2. in experiment 1, the birds receiving the 0, 75, 100, 125 mg probiotic/kg diets had weight gains of 1204.0, 1272.0, 1268.3 and 1210.5, respectively at the end of 8 weeks of feeding. the group of birds fed on the 75 mg probiotic supplemented diet retained significantly (p < 0.01) more nitrogen than the control birds. serum cholesterol ...19968773848
isolation and characterization of the nuclease o gene (nuco) from aspergillus oryzae.nuclease o in the mycelia of aspergillus oryzae has been purified 55-fold by successive steps of chromatography from the filtrate of the autolyzate. the molecular mass of nuclease o was 32 kda, as estimated by sds polyacrylamide-gel electrophoresis. the nuclease o gene (nuco) encoding this enzyme was cloned and sequenced. the open reading frame is interrupted by four introns with conserved splice sites and contains 328 amino-acid residues of the mature enzyme. a. nidulans transformants obtained ...19968781174
fungal contamination of potential medical interest in spanish grain stores.a one-year study was made of fungal spores detected in the air and in the grain of two silos and two seed stores near córdoba, spain. gravimetric and volumetric methods were used simultaneously on culture mediums to sample the air. the dilution method was employed to analyze seed contamination. a total of 70 taxa were isolated, 67 of these from the air and 46 in seeds. the most abundant airborne taxa were: aspergillus oryzae, cladosporium cladosporioides and yeast, while yeast, a. niger and a. o ...19968807511
cloning and nucleotide sequence of the mono- and diacylglycerol lipase gene (mdlb) of aspergillus oryzae.aspergillus oryzae ifo4202 produces at least two extracellular lipolytic enzymes l1 and l2 (cutinase, and mono- and diacylglycerol lipase, respectively). southern hybridization of restriction enzyme-digested genomic dna fragments with 23-mer oligonucleotides synthesized according to the amino acid sequence of the l2 as probe suggested the presence of the l2 gene (tentatively designated as mdlb) and an additional weakly hybridizing region. a fragment containing the genomic mdlb gene was cloned in ...19968807803
ergot alkaloid glycosides with immunomodulatory activities.new glycosides derived from ergot alkaloids elymoclavine and dh-lysergol were synthesized by chemoenzymatic methods. beta-glucosides were obtained either by chemical method or by transglycosylation (glycosidase from aspergillus oryzae), lactosides were prepared by further extension of carbohydrate chain using beta-1,4-galactosyltransferase (bovine milk) and alpha-5-n-acetylneuraminyl-(2-->6)-beta-d-galactopyranosyl-(l-->4)-2- acetamido-2-deoxy-beta-d-glucopyranosyl-(1-->o)-elymoclavine was prepa ...19968818236
acceleration of sterol excretion, little meteorism, and less inhibition of iron absorption by okara koji, a new foodstuff, in rats.okara koji (ok) is the term for okara fermented by aspergillus oryzae (a fungus for making miso koji). the amount of sterol excreted in the feces from rats fed with ok as the dietary fiber (df) source was greater than that from rats fed with okara (oc, the insoluble residue of homogenized soybean) as the df source. the digestive residue of ok's insoluble dietary fiber (idf) was more porous than that of oc-idf. the stronger sterol excretion with ok would have arisen from the complementary action ...19968829521
heating inactivates the enzymatic activity and partially inactivates the allergenic activity of asp o 2.sensitization to various flours and flour additives in the baking industry has been known for some time. however, most studies refer to allergens in their native state.19968835132
raw-starch-digesting and thermostable alpha-amylase from the yeast cryptococcus sp. s-2: purification, characterization, cloning and sequencing.a starch-degrading enzyme produced by the yeast cryptococcus sp. s-2 was purified in only one step by using an alpha-cyclodextrin-sepharose 6b column, and was characterized as an alpha-amylase (ec 3.2.1.1). the molecular mass and isoelectric point of purified alpha-amylase (amy-cs2) were estimated to be 66 kda and 4.2 respectively. amy-cs2 has raw-starch-digesting and raw-starch-absorbing activities. furthermore it was shown to be thermostable. an open reading frame of the cdna specified 611 ami ...19968836148
characterization of soybean allergens causing sensitization of occupationally exposed bakers.fourteen bakers suffering from workplace-related respiratory symptoms and sensitized to soybean were studied. twelve of them were also allergic to wheat flour, 10 to rye flour, and five to alpha-amylase of aspergillus oryzae (asp o 2). ige estimation by rast strongly indicated that the trypsin inhibitor and lipoxidase are major allergens of soybean. various allergenic components could be characterized by immunoblotting after two-dimensional electrophoresis. our rast and immunoblotting results sh ...19968836337
influence of gelatin matrices cross-linked with transglutaminase on the properties of an enclosed bioactive material using beta-galactosidase as model system.transglutaminase (protein-glutamine: amine gamma-glutamyltransferase, ec 2.3.2.13) from streptoverticillium mobaraense has been used to stabilize immobilisates produced with beta-galactosidase (beta-d-galactoside galactohydrolase, ec 3.2.1.23) from aspergillus oryzae and acid-processed gelatins of different qualities as support. the isopeptide level of n epsilon-(gamma-l-glutamyl)-l-lysine bonds formed by transglutaminase was determined to estimate their influence on the kinetic properties of th ...19968853118
the synthesis of galactopyranosyl derivatives with beta-galactosidases of different origins.beta-galactosidases from four different sources were used to catalyze the transfer of beta-d-galactopyranosyl from 4-nitrophenyl-beta-d-galactopyranoside to a hydroxyl group of 2-acetamido-2-deoxy-galactopyranose in the synthesis of gal beta (1-3)galnac (1), gal beta (1-4)galnac (2) and gal beta (1-6)galnac (3), in triethyl phosphate buffered solutions (20-60%). when beta-galactosidases from penicillium multicolor and aspergillus oryzae were used as the catalysts, the beta (1-6)-linked disacchar ...19968870243
the effect of 6'-galactooligosaccharides on bone mineralization of rats adapted to different levels of dietary calcium.6'-galactooligosaccharides (6'-gos), a mixture of galactosyl oligosaccharides formed from lactose by the transgalactosyl reaction with beta-d-galactosidase derived from aspergillus oryzae and streptococcus thermophillus, are unhydrolyzed in the small intestine and are fermented by the intestinal bacteria. the effects of 6'-gos on calcium (ca) absorption and bone mineralization were examined in male wistar rats adapted to different levels of dietary ca for 30 days. dietary 6'-gos (5 g/100 g of di ...19968899459
construction of a promoter probe vector autonomously maintained in aspergillus and characterization of promoter regions derived from a. niger and a. oryzae genomes.we used a plasmid carrying a sequence for autonomous maintenance in aspergillus (ama1) and the e. coli uida gene as a reporter gene to search the a. oryzae and a. niger genomes for dna fragments having strong promoter activity. beta-glucuronidase (gus)-producing a. oryzae transformants containing the no. 8an derived from a. niger, or the no. 9ao derived from a. oryzae, were constitutive for the expression of the uida gene when cultivated in the presence of a variety of carbon and nitrogen source ...19968901095
cloning and sequencing of the gene encoding tannase and a structural study of the tannase subunit from aspergillus oryzae.we cloned the aspergillus oryzae tannase gene using three oligodeoxyribonucleotide (oligo) probes synthesized according to the tannase n-terminal and an internal amino acid (aa) sequence. the nucleotide (nt) sequence of the tannase gene was determined and compared with that of a tannase dna complementary to rna (cdna) by means of reverse transcriptase pcr. the results indicated that there was no intron in the tannase gene and that it coded for 588 aa with a molecular weight of about 64,000. the ...19968917102
pectin methyl esterase from aspergillus aculeatus: expression cloning in yeast and characterization of the recombinant enzyme.seventeen full-length cdnas encoding pectin methyl esterase i (pme i) have been isolated from the filamentous fungus aspergillus aculeatus by expression cloning in yeast. yeast colonies expressing functional pme i were identified on agar plates containing highly esterified pectin, and a cdna encoding pme i was isolated. the deduced amino acid sequence of pme i is highly similar (74% identity) to the pme from aspergillus niger. a full-length cdna encoding pme i was cloned into an aspergillus expr ...19968920970
deletion analysis of promoter elements of the aspergillus oryzae agda gene encoding alpha-glucosidase.the nucleotide sequence of a 1.5-kb fragment of the promoter region of the aspergillus oryzae agda gene encoding alpha-glucosidase was determined. a comparison with the promoter regions of other aspergillus amylase genes indicated that there are three highly conserved sequences, designated regions i, ii and iii, located at -670 nt, -596 nt and -544 nt relative to the start codon, respectively. the function of these consensus sequences in the agda promoter was investigated by deletion analysis of ...19968929396
separation of enzymes from polyenzyme mixtures used in medicine and pharmacy. i. isolation and characterization of xylanase from aspergillus oryzae.three multiple forms (f-i, f-ii and f-iii) of xylanase produced from aspergillus oryzae were established in the commercial preparation luizym. two methods were used (ion-exchange chromatography and polyacrylamide gel electrophoresis). the active xylanase fraction (f-iii-ii-i) was purified and characterized (phopt 4.5; topt 37 degrees c and molecular mass 38.5 kda). the products of the enzymatic reaction were analysed by hplc. the hydrolysis pattern showed that the xylanase was an exo-endo acting ...19968941946
method for the extraction of riboflavin for high-performance liquid chromatography and application to casein.three different approaches, including the commonly used acid digestion, were compared for the efficient extraction of very low concentrations of riboflavin (vitamin b2) from casein. an extraction method that used pepsin to release riboflavin was the most efficient. the pepsin extraction method was then further optimized using a factorial design to yield the maximum amount of riboflavin. the enzyme takadiastase was used for the conversion of all forms of riboflavin to the free form, and this conv ...19968952454
purification, characterization, molecular cloning, and expression of two laccase genes from the white rot basidiomycete trametes villosa.two laccases have been purified to apparent electrophoretic homogeneity from the extracellular medium of a 2,5-xylidine-induced culture of the white rot basidiomycete trametes villosa (polyporus pinsitus or coriolus pinsitus). these proteins are dimeric, consisting of two subunits of 63 kda as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and have typical blue laccase spectral properties. under nondenaturing conditions, the two purified laccases have different pis; pur ...19968975613
efficient expression of a phanerochaete chrysosporium manganese peroxidase gene in aspergillus oryzae.a manganese peroxidase gene (mnp1) from phanerochaete chrysosporium was efficiently expressed in aspergillus oryzae. expression was achieved by fusing the mature cdna of mnp1 with the a. oryzae taka amylase promoter and secretion signal. the 3' untranslated region of the glucoamylase gene of aspergillus awamori provided the terminator. the recombinant protein (rmnp) was secreted in an active form, permitting rapid detection and purification. physical and kinetic properties of rmnp were similar t ...19968975615
new international f.i.p. method for the determination of the activity of aspergillus oryzae proteases.proteases of aspergillus oryzae are used as a drug in the therapy of digestive disorders. to standardize these enzyme the enzyme commission of the fédération internationale pharmaceutique (f.i.p.) has tested a new determination method, which will be described below. the standard preparation of a mixture of aspergillus oryzae proteases used in this test is characterized.19968985984
cloning, sequencing, and expression of the cellulase genes of humicola grisea var. thermoidea.we have cloned an endoglucanase (egi) gene and a cellobiohydrolase (cbhi) gene of humicola grisea var. thermoidea using a portion of the trichoderma reesei endoglucanase i gene as a probe, and determined their nucleotide sequences. the deduced amino acid sequence of egi was 435 amino acids in length and the coding region was interrupted by an intron. the egi lacks a hinge region and a cellulose-binding domain. the deduced amino acid sequence of cbhi was identical to the h. grisea cbhi previously ...19968987622
sequence-specific binding sites in the taka-amylase a g2 promoter for the crea repressor mediating carbon catabolite repression.the n-terminal part of the crea protein encompassing two zinc fingers was expressed in escherichia coli as a fusion protein with the maltose binding protein (male) of e. coli. our results show that crea binds to the promoter of the taa-g2 gene encoding taka-amylase a of aspergillus oryzae. dnase i footprinting experiments showed that crea bound to three sites with high affinity and to one site with low affinity within the first 401-bp region upstream of the transcription initiation site. all of ...19968987852
molecular cloning, purification and characterization of two endo-1,4-beta-glucanases from aspergillus oryzae kbn616.two endo-1,4-beta-glucanase genes, designated cela and celb, from a shoyu koji mold aspergillus oryzae kbn616, were cloned and characterized. the cela gene comprised 877 bp with two introns. the cela protein consisted of 239 amino acids and was assigned to the cellulase family h. the celb gene comprised 1248 bp with no introns. the celb protein consisted of 416 amino acids and was assigned to the cellulase family c. both genes were overexpressed under the promoter of the a. oryzae taka-amylase a ...19969008887
[uses of microbial beta-galactosidases to reduce lactose content in milk and dairy products].the commercial sources of microbial beta-galactosidases (lactases) include the yeasts species kluyveromyces marxianus, kluyveromyces lactis and candida kefyr which are used to hydrolyse lactose in milk due to their optimum ph. on the other hand, lactases obtained from the moulds aspergillus niger and aspergillus oryzae have an acid optimum ph and therefore are used to hydrolyse lactose in whey to obtain whey syrups to be used as raw materials in the food industry. the lactose intolerance problem ...19969122548
interaction of proteases with legume seed inhibitors. molecular features.after having found that raw black beans (phaseolus vulgaris) were toxic, while the cooked ones constitute the basic diet of the underdeveloped peoples of the world, in the sixties, our research directed by dr. jaffé, concentrated mainly around the detection and identification of the heat labile toxic factors in legume seeds. a micromethod for the detection of protease inhibitors (pi) in individual seeds was developed, for the purpose of establishing that the multiple trypsin inhibitors (ti) foun ...19969137634
molecular cloning of a cdna encoding enolase from the filamentous fungus, aspergillus oryzae.a 1.6-kbp full-length cdna for the aspergillus oryzae enolase gene (enoa) was cloned. the sequenced insert contained a continuous open reading frame of 1314 bp encoding a protein of molecular weight 47 405. among all enolases sequenced to-date, the deduced amino-acid sequence showed the highest homology (74.9%) with candida albicans enolase (eno1). strong codon biases and multiple transcription start sites downstream from ct-blocks in the 5'-flanking region suggested strong expression. enoa mrna ...19968929395
molecular cloning of a genomic dna for enolase from aspergillus oryzae.we have isolated an enolase gene (enoa) from aspergillus oryzae by heterologous hybridization using the corresponding saccharomyces cerevisiae eno2 gene as a probe. a 2.9-kb bglii-fragment contained the entire structural gene enoa including 5'- and 3'- flanking regions. the homology between a. oryzae enoa and s. cerevisiae eno2 genes is 66.9% when introns are removed. genomic southern analysis indicated that there is only one enolase gene in a. oryzae.19968824839
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
the crystal structure of ribonuclease rh from rhizopus niveus at 2.0 a resolution.the three-dimensional structure of ribonuclease rh (rnase rh), a new class of microbial ribonuclease from rhizopus niveus, has been determined at 2.0 a resolution. the overall structure of rnase rh is completely different from those of other previously studied rnases, such as rnase a from bovine pancreas and rnase t1 from aspergillus oryzae. in the structure of rnase rh, two histidine residues (his46 and his109) and one glutamic acid residue (glu105), which were predicted to be critical to the a ...19968551522
the identification and characterization of four laccases from the plant pathogenic fungus rhizoctonia solani.four distinct laccase genes, lcc1, lcc2, lcc3 and lcc4, have been identified in the fungus rhizoctonia solani. both cdna and genomic copies of these genes were isolated and characterized. hybridization analyses indicate that each of the four laccase genes is present in a single copy in the genome. the r. solani laccases can be divided into two groups based on their protein size, intron/exon organization, and transcriptional regulation. three of these enzymes have been expressed in the fungus asp ...19968598061
safety evaluation of a lipase expressed in aspergillus oryzae.a programme of studies was conducted to establish the safety of a lipase artificially expressed in aspergillus oryzae to be used in the detergent industry and as a processing aid in the baking industry. laboratory animal studies were used to assess general and inhalation toxicity, skin sensitization, and skin and eye irritation. its potential to cause mutagenicity and chromosomal aberrations was assessed in microbial and tissue culture in vitro studies. the pathogenicity of a. oryzae, the organi ...19968606032
morphology and physiology of an alpha-amylase producing strain of aspergillus oryzae during batch cultivations.the microscopic morphology, that is, total hyphal length and total number of tips, has been characterized during batch cultivations of aspergillus oryzae. the specific growth rate estimated by measuring the total hyphal length (mu(h)) corresponds well with the specific growth rate estimated from dry weight measurements during cultures grown as free hyphal elements. the average tip extension rate can be described with a saturation type kinetics with respect to the average total hyphal length, and ...199618623577
effect of organic solvents on enantioselectivity of protease catalysis.the protease-catalyzed transesterifications between n-trifluoroacetyl-dl-phenylalanine 2,2,2-trifluoroethyl ester and 1-propanol were studied in a variety of anhydrous organic solvents at 30 degrees c. the protease preparations lyophilized from phosphate buffer solutions (ph 8.0) were used as catalysts. the organic solvent affected both rate of reaction and enantioselectivity differently. proteases such as aspergillus oryzae protease, subtilisin carlsberg, and subtilisin bpn' always preferred th ...199718629956
proteolysis in dry fermented sausages: the effect of selected exogenous proteases.the effect of three commercial proteases (pronase e from streptomyces griseus, aspartyl proteinase from aspergillus oryzae and papain) on protein breakdown and the sensory characteristics of dry fermented sausages was investigated. water soluble, non-protein, 5% phosphotungstic acid soluble, 5% sulphosalicylic acid soluble and total volatile basic nitrogen contents increased during fermentation, stabilizing later until the end of ripening (26th day). nitrogen values were always greater in the as ...199722061850
isolation of taka-amylase a peptides required for substrate binding.an α-amylase from aspergillus oryzae, taka-amylase a (taa), was cleaved into peptide fragments by an acid protease. inactivation of taa was greatly retarded by the addition of α-cyclodextrin or ca(2+). taa peptide fragments were separated into two groups having no and high affinity to the substrate, soluble starch. this separation was done by the forced affinity chromatography method by a column of epichlorohydrin cross-linked soluble starch gel. three peptides were isolated from the high-affini ...199727396736
[comparative characteristics of microbial proteases by the level of hydrolysis of protein substrates].screening of enzyme preparations displaying a maximum proteolytic activity at ph 4.0-5.5 and effecting deep proteolysis of plant proteins was performed. amyloprotooryzin prepared from aspergillus oryzae 387 containing a complex of proteolytic enzymes was the most effective. the amino acid composition of the hydrolysates obtained was studied. amyloprotooryzin increased the contents of amino acids by 108-227%, depending on the substrate used. the enzymatic complex of amyloprotooryzin was studied; ...19979139311
safety evaluation of tannase enzyme preparation derived from aspergillus oryzae.tannase is an acylhydrolase enzyme preparation from aspergillus oryzae that can be used as a processing aid for the manufacture of cold water-soluble tea beverages. a 91-day oral toxicity study in the rat and a gene mutation study in salmonella typhimurium were performed to establish the safety of the enzyme preparation for the consumer. general toxicity was low, with no adverse effects observed at the highest dose tested, 1% in the diet. there was no evidence of mutagenic potential with or with ...19979146733
effects of lactitol-oligosaccharides on calcium and magnesium absorption in rats.lactitol-oligosaccharide (lo) was prepared from lactitol by transglycosylation reaction with aspergillus oryzae beta-galactosidase. lo is resistant to metabolism in the small intestine but not in the large intestine. the effects of lo, lactose (lac), lactitol (lacol) and galactooligosaccharide (gl) on calcium and magnesium absorption were determined by feeding 8-week-old sprague-dawley male rats diets containing 5% of the above carbohydrates for two weeks. the results obtained were as follows. 1 ...19979151246
cloning of a protopectinase gene of trichosporon penicillatum and its expression in saccharomyces cerevisiae.a protopectinase (ppase)-encoding gene, pse3, from trichosporon penicillatum was cloned by colony hybridization using two oligonucleotide probes synthesized from the n-terminal amino acid sequences of native ppase se1 and one peptide from a lysyl endopeptidase digest. nucleotide sequencing revealed that pse3 contains an orf encoding a 367 amino acid protein. mature ppase se3 is composed of 340 amino acids and the n-terminus of the orf appeared to correspond to a signal peptide and a propeptide p ...19979168614
molecular cloning and heterologous expression of the isopullulanase gene from aspergillus niger a.t.c.c. 9642.isopullulanase (ipu) from aspergillus niger a.t.c.c. (american type culture collection) 9642 hydrolyses pullulan to isopanose. ipu is important for the production of isopanose and is used in the structural analysis of oligosaccharides with alpha-1,4 and alpha-1,6 glucosidic linkages. we have isolated the ipua gene encoding ipu from the filamentous fungi a. niger a.t.c.c. 9642. the ipua gene encodes an open reading frame of 1695 bp (564 amino acids). ipu contained a signal sequence of 19 amino ac ...19979169610
efficient expression of mono- and diacylglycerol lipase gene from penicillium camembertii u-150 in aspergillus oryzae under the control of its own promoter.the gene, mdla, coding for mono- and diacylglycerol lipase from penicillium camembertii u-150 was expressed efficiently in aspergillus oryzae under the control of its own promoter. the gene product was secreted into the culture medium with a highest productivity of 1 g/liter and correctly processed at both n- and c-termini. kex2-like processing was suggested to occur at the c-terminus in both a. oryzae and p. camembertii. specific activity and substrate specificity of the purified recombinant pr ...19979178556
the acetate regulatory gene facb of aspergillus nidulans encodes a zn(ii)2cys6 transcriptional activator.genetic studies have indicated that the facb gene of aspergillus nidulans is a major regulatory gene involved in acetamide and acetate utilisation. sequencing of the facb gene revealed that it encodes a protein that contains an n-terminal gal4-like zn(ii)2cys6 (or c6 zinc) binuclear cluster for dna binding, leucine zipper-like heptad repeat motifs and central and c-terminal acidic alpha-helical regions, consistent with a function as a dna-binding transcriptional activator. the zn(ii)2cys6 cluste ...19979197408
influence of tallow and aspergillus oryzae fermentation extract in dairy cattle rations.objectives were to determine the effects of adding 3 g/d of aspergillus oryzae fermentation extract to diets with or without 5.6% added tallow. twenty-eight holstein cows (mean = 98 d of lactation) were assigned to a randomized block experiment in a 2 x 2 factorial arrangement of treatments. treatments were the basal diet 1) without tallow or extract, 2) with extract but no tallow, 3) with tallow but no extract, and 4) with tallow and extract. milk production, dry matter intake, 3.5% fat correct ...19979201589
an overview of the safety evaluation of the thermomyces lanuginosus xylanase enzyme (sp 628) and the aspergillus aculeatus xylanase enzyme (sp 578).xylanases sp 628 and sp 578 were produced by submerged fermentation of aspergillus oryzae, containing a gene code originating from thermomyces lanuginosus and aspergillus aculeatus, respectively. both enzymes were subject to the same series of toxicological tests to document their safety in use. the enzymes are to be applied as processing aids in the baking industry and in wheat starch separation. neither enzyme was found to be mutagenic in the salmonella typhimurium reverse mutation assay, nor ...19979205568
characterization of the gene encoding an extracellular laccase of myceliophthora thermophila and analysis of the recombinant enzyme expressed in aspergillus oryzae.a genomic dna segment encoding an extracellular laccase was isolated from the thermophilic fungus myceliophthora thermophila, and the nucleotide sequence of this gene was determined. the deduced amino acid sequence of m. thermophila laccase (mtl) shows homology to laccases from diverse fungal genera. a vector containing the m. thermophila laccase coding region, under transcriptional control of an aspergillus oryzae alpha-amylase gene promoter and terminator, was constructed for heterologous expr ...19979251203
structure of the aspergillus oryzae alpha-amylase complexed with the inhibitor acarbose at 2.0 a resolution.the three-dimensional structure of the aspergillus oryzae alpha-amylase (taka-amylase), in complex with the inhibitor acarbose, has been determined by x-ray crystallography at a resolution of 1. 98 a. the tetrasaccharide inhibitor is present as a hexasaccharide presumably resulting from a transglycosylation event. the hexasaccharide occupies the -3 to +3 subsites of the enzyme, consistent with the known number of subsites determined by kinetic studies, with the acarbose unit itself in the -1 to ...19979283074
purification and characterization of an aspergillus oryzae-produced carboxylesterase that catalyzes o-deacetylation of a fully acetylated o-glucoside of n-phenylacetohydroxamic acid.a carboxylesterase [2,3,4,6-tetra-o-acetyl-1-[(n-acetyl-n-phenylamino)oxy]-1-deoxy-beta-d-g lucopyranoside (gpa) o-deacetylase] from a culture product of aspergillus oryzae (taka diastase) was purified 8500-fold with a yield of 3%. the molecular mass of the purified enzyme was shown to be 35 +/- 1 kda by sds/page. the enzyme shows a selective o-deacetylation activity of gpa to give the fully o-deacetylated glucoside. among the substrates tested, the enzyme did not hydrolyze benzoyl and phenylace ...19979310360
tmc-2a, -2b and -2c, novel dipeptidyl peptidase iv inhibitors produced by aspergillus oryzae a374. i. taxonomy of producing strain, fermentation, and biochemical properties.tmc-2a(1), -2b (2) and -2c (3), novel dipeptidyl peptidase iv (dpiv) inhibitors, were isolated from the fermentation broth of aspergillus oryzae a374. tmc-2a, -2b and -2c inhibited rat kidney dpiv with ic50 value of 8.1 microm, 17 microm, and 20 microm, respectively, as well as human dpiv prepared from mononuclear cells and adenocarcinoma cells. tmc-2 compounds inhibited only dpiv among the proteases tested, indicating their high selectivity for dpiv. the kinetic analyses revealed that tmc-2a wa ...19979315076
tmc-2a, -2b and -2c, new dipeptidyl peptidase iv inhibitors produced by aspergillus oryzae a374. ii. isolation and structure determination.new dipeptidyl peptidase iv inhibitors, tmc-2a, -2b, and -2c, were isolated from the fermentation broth of aspergillus oryzae a374. on the basis of chemical, spectroscopic and x-ray crystallographic analyses, their structures were established to be peptide-like compounds composed of three moieties, l-tryptophan, mono- or dihydroxy-l-leucine and highly substituted isoquinoline.19979315077
optical resolution of dl-alanine by using immobilized aspergillus oryzae cells.mycelium pellets with diameters of 1-2 mm and containing abundant aminoacylase were obtained by the liquid fermentation of aspergillus oryzae 3042. by the cross-linking method with reagents of gelatin and formaldehyde, the immobilized a. oryzae cells (iac) were prepared with much higher activity and reactive properties. the effects of factors on enzymatic activity of iac were investigated. when substrate concentration was less than 0.15 mol/l, the michaelis-menten mechanism was suitable for this ...19979343710
cloning and sequence analysis of the gene (epra1) encoding an extracellular protease from aeromonas hydrophila.a gene (epra1) encoding the extracellular protease of aeromonas hydrophila ah1 has been cloned and sequenced. nucleotide sequence analysis of epra1 predicted a single open reading frame (orf) of 1038 bp encoding a 346 amino acid (aa) polypeptide, with a potential 21-aa signal peptide. when the epra1 gene was expressed in minicells, one major band of approx. 37 kda was identified, while protease activity staining experiments identified a caseinolytic band of approx. 29 kda determined by sds-page ...19979358060
effect of steam-flaked or steam-rolled corn with or without aspergillus oryzae in the diet on performance of dairy cows fed during hot weather.the objective of this study was to determine the effects of steam-rolled versus steam-flaked corn in the diet with or without the addition of a culture of aspergillus oryzae on the performance of high producing dairy cows during hot summer weather. thirty-two holstein cows averaging 92 (+/- 60) d in milk were fed a pretreatment diet for 21 d followed by a 70-d experimental period in a completely randomized block design with a 2 x 2 factorial arrangement of treatments. diets were 1) steam-flaked ...19979436111
in vivo antioxidant activity of okara koji, a fermented okara, by aspergillus oryzae.the antioxidants in okara koji (ok), an okara (oc) fermented by aspergillus oryzae, gamma-tocopherol, delta-tocopherol, genistin, daizein, genistein, and 3-hydroxyanthranilic acid were identified by hplc. ok's extract with 80% methanol strongly inhibited linoleate peroxidation, much more than other ok's extracts with hexane or hot water. the methanol extract accelerated 12-hydroxyeicosatetraenoic acid formation in membrane lipids at 10(-3) concentration, but inhibited the formation at higher con ...19979438976
immobilization of enzymes on methacrylamide-based copolymers.a series of reactive polymeric supports for the covalent binding of enzymes was prepared from methacrylamide in conjunction with various copolymerized monomers. in all cases the specific surface area of the prepared copolymer particles increased with the ratio of crosslinker to monomer. the polymers containing oxirane and aldehyde groups were promising for direct binding of beta-galactosidase, whereas the supports containing amino groups also qualified for enzyme immobilization with glutaraldehy ...19979090722
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