investigation of the functional interplay between the primary site and the subsite of rnase t1: kinetic analysis of single and multiple mutants for modified substrates.we report on the functional cooperativity of the primary site and the subsite of ribonuclease t1 (rnase t1; ec the kinetic properties of the single tyr-38-phe and asn-98-ala mutants have been compared with those of the corresponding double mutant. the tyr-38-phe mutation has been used to probe enzyme-substrate interactions at the primary site; the asn-98-ala mutation monitors subsite interactions. in addition to the dinucleoside phosphate substrate gpc, we measured the kinetics for gp ...19948208724
[are amylases in bakery products and flour potential food allergens?].the enzyme alpha-amylase from the mould aspergillus oryzae (asp o ii) routinely used for the production of bread, cakes and pastries has in recent years been identified as an inhalative allergen for occupational diseases (bakers' asthma). it is doubtful whether this amylase in the final product, i.e. after the baking procedure, can still be regarded as an allergen. to clarify this question, detailed case histories on 138 subjects were recorded (98 allergics, 20 patients suffering form chronic in ...19948209207
probiotics and manipulation of rumen development and function.there is good evidence that the bacterial and fungal probiotics are effective in the manipulation of rumen development and function. the effects of adherent streptococcus bovis preparations were manifested by significant increase of adherent s. bovis bacteria and alpha amylase activity in the rumen wall of young ruminants. fungal probiotics based on saccharomyces cerevisiae and aspergillus oryzae are rapidly gaining acceptance as a means of improving productivity in adult ruminants. however, fun ...19938215880
effects of an aspergillus oryzae fermentation extract and other factors on lactate utilization by the ruminal bacterium megasphaera elsdenii.the objective of this study was to determine the effects of an aspergillus oryzae fermentation extract (amaferm) as well as other factors on lactate utilization by the ruminal bacterium megasphaera elsdenii b159. addition of amaferm or a filter-sterilized amaferm filtrate stimulated l-lactate uptake by both m. elsdenii and the ruminal selenomonad strain h18. growth of m. elsdenii in medium that contained dl-lactate (2 g/l), trypticase, and yeast extract was only slightly stimulated by the additi ...19938226379
miso from peas (pisum sativum) and beans (phaseolus vulgaris) of domestic origin. fermented foods from agricultural products in europe. ii.miso is a fermented soybean paste widely used in japan as a soup base or as a seasoning agent. koji (cereal grains with the mold aspergillus oryzae) serves as enzyme source. peas (pisum sativum) and beans (phaseolus vulgaris) of german origin can be used as substitutes for soybeans in the preparation of miso-like products. the legumes (peas, beans and soybeans for comparison) are washed, soaked in boiled water, dehulled and cooked for 35 min at reduced pressure. after grounding the seeds are mix ...19938237083
characterization of an aspartic proteinase of mucor pusillus expressed in aspergillus oryzae.the aspartic proteinase (mpp) gene from the zygomycete fungus mucor pusillus was introduced into an ascomycete fungus, aspergillus oryzae, by protoplast transformation using the nitrate reductase (niad) gene as the selective marker. southern blot analysis indicated that the mpp gene was integrated into the resident niad locus at a copy number of 1-2. mpp secreted by the recombinant a. oryzae was correctly processed but was more highly glycosylated than that produced in the original m. pusillus s ...19938246885
the sequence and x-ray structure of the trypsin from fusarium oxysporum.the trypsin from fusarium oxysporum is equally homologous to trypsins from streptomyces griseus, streptomyces erythraeus and to bovine trypsin. a dfp (diisopropylfluorophosphate) inhibited form of the enzyme has been crystallized from 1.4 m na2so4, buffered with citrate at ph 5.0-5.5. the crystals belong to space group p2(1) with cell parameters a = 33.43 a, b = 67.65 a, c = 39.85 a and beta = 107.6 degrees. there is one protein molecule in the asymmetric unit. x-ray diffraction data to a resolu ...19938332590
preparation, properties and application of aspergillus oryzae s1 nuclease covalently bound to aminobutyl-bio-gel p-2 through its carbohydrate moiety.purified aspergillus oryzae s1 nuclease, when covalently coupled to aminobutyl-(ab)-bio-gel p-2, via its carbohydrate moiety, retained 40-50% activity of the soluble enzyme. optimization of coupling conditions showed that the most active immobilized preparations are obtained when 50-60 units of 1 mm periodate-oxidized enzyme are allowed to react with 1 ml (packed volume) of ab-bio-gel p-2 at 4 degrees c, in the presence of 20% (v/v) ethylene glycol, for 15 h. immobilization did not change the ph ...19938338642
secondary structure and temperature-induced unfolding and refolding of ribonuclease t1 in aqueous solution. a fourier transform infrared spectroscopic study.the secondary structure of ribonuclease t1 (rnase t1) in aqueous solution and its temperature-induced structural changes have been investigated by fourier-transform infrared (ft-ir) spectroscopy. 13 to 14% alpha-helix and 32 to 33% beta-sheet were estimated from the resolution-enhanced ft-ir spectra, in agreement with the crystal structure which indicates 16% alpha-helix and 35% beta-sheet. specific ir-marker bands are assigned to the different beta-sheet structures, to the slightly bent alpha-h ...19938355280
structural features of a polygalacturonase gene cloned from aspergillus oryzae kbn616.a genomic gene encoding a polygalacturonase from aspergillus oryzae, used in soy sauce production, was cloned and sequenced. the structural gene comprises 1227 bp coding for 363 amino acids with a putative prepropeptide of 28 amino acids and the open reading frame is disrupted by two short introns of 57 bp and 81 bp. the deduced amino acid sequence of the mature protein showed 63, 63, 63 and 64% homology with those of aspergillus niger polygalacturonase i, aspergillus niger polygalacturonase ii, ...19938359678
polypeptide folding of bacillus cereus atcc7064 oligo-1,6-glucosidase revealed by 3.0 a resolution x-ray analysis.the crystal structure of an oligo-1,6-glucosidase from bacillus cereus atcc7064 was determined by the x-ray diffraction method at 3.0 a resolution. the structure was solved by the multiple isomorphous replacement method and refined to a crystallographic r-factor of 0.208, using the molecular dynamics refinement program, x-plor. the electron density map revealed the folding of a polypeptide chain consisting of 558 amino acid residues. the molecule can be subdivided into three domains (n-terminal ...19938370659
effects of aspergillus oryzae fermentation extract on in vitro equine cecal fermentation.the objective of this study was to examine the effects of aspergillus oryzae fermentation extract on the in vitro equine cecal fermentation of soluble starch, amino acids/peptides, coastal bermudagrass hay, and alfalfa hay. cecal contents were obtained from a cecally fistulated quarter horse gelding fed coastal bermudagrass and grain (70:30) either unadapted or adapted to dietary a. oryzae supplementation (2 g/d). mixed cecal microorganisms were incubated in anaerobic media for either 24 h (solu ...19938376241
influence of aspergillus oryzae fermentation extract on forage intake, site of digestion, in situ degradability, and duodenal amino acid flow in steers grazing cool-season pasture.ten ruminally and duodenally cannulated (326 +/- 28 kg) and four esophageally fistulated (394 +/- 23 kg) steers grazing cool-season pasture throughout the growing season were used to evaluate the influence of aspergillus oryzae fermentation extract (ao) supplementation on intake, forage nutrient utilization, and duodenal amino acid flow. steers grazed a predominantly smooth brome (bromus inermis l.) pasture, and measurements were taken in three periods (june, july, and august). steers were dosed ...19938385089
effect of aspergillus oryzae fermentation extract (amaferm) on in vitro fiber degradation.the influence of aspergillus oryzae fermentation extract (amaferm) on in vitro fiber degradation was determined by incubating eight ground fibrous feed-stuffs with rumen fluid and buffer inoculum. amaferm was added at 0, .4, .8, or 1.2 g/l of fermentation mixture. both ndf and adf degradabilities were determined after 96 h of incubation. addition of extract had no effect on ndf or adf degradability of pure cellulose, low endophyte fescue, wheat straw, corn silage, or prairie hay. addition of ama ...19938385163
effect of nonfiber carbohydrate level and aspergillus oryzae fermentation extract on intake, digestion, and milk production in lactating dairy cows.eight multiparous, ruminally cannulated holstein cows averaging 40 d in milk and 575 kg bw at the start of trial were in a replicated 4 x 4 latin square arrangement (28-d periods) to determine the effects of dietary nonfiber carbohydrate (nfc) level and aspergillus oryzae fermentation extract (ao) on intake, milk production, and nutrient digestibility. treatments were 42 or 35% nfc and 0 or 3 g of ao per day arranged as a 2 x 2 factorial. diets formulated to contain 21% ndf from alfalfa silage ( ...19938386716
crystallization and x-ray diffraction analysis of recombinant coprinus cinereus peroxidase.crystals suitable for an x-ray diffraction investigation have been obtained of recombinant coprinus cinereus peroxidase expressed in aspergillus oryzae. the crystals were grown by the hanging drop method with polyethylene glycol 6000 as the precipitant. a ph range from 6.2 to 8.0 and cacl2 or mgcl2 present at a concentration of 0.35 m were essential for the crystal growth. a metastable monoclinic modification can be obtained under certain conditions, and with variations in temperature they are t ...19938394939
expression of aspergillus oryzae alpha-amylase gene in saccharomyces cerevisiae.a fragment containing the full length cdna from aspergillus oryzae alpha-amylase has been amplified by pcr using specific synthetic oligonucleotides. the amplified cdna was designed to favour its expression in yeast by modifying its upstream untranslated region. it was subcloned in the expression vector pyex alpha 1, placed under the control of the yeast cyc1-gal10 promoter and used to transform saccharomyces cerevisiae. cells were then able to express and secrete active alpha-amylase to the med ...19938405943
antibodies against active-site peptides common to glucosyltransferases of mutans streptococci.polyclonal antibodies were raised against peptides derived from an active-site sequence common to the family of mutans streptococcal glucosyltransferases (gtfs). the sequence contains an aspartic acid residue that functions in formation of the enzyme transition state in catalysis. two gtfs were targeted with similar but not identical sequences in this region: one that synthesizes an alpha-1,3-linked water-insoluble glucan and a homologous gtf that synthesizes an alpha-1,6-linked water-soluble gl ...19938406882
decreased protein-stabilizing effects of cryoprotectants due to crystallization.the stabilizing effects of various additives against inactivation of an enzyme (beta-galactosidase from aspergillus oryzae) during freeze-drying were studied, with a focus on their crystallinity. the crystalline morphology of mannitol and inositol in freeze-dried cakes depended on the solute concentrations before freezing and the freeze-drying method used. the additives in their amorphous state showed concentration-dependent stabilization of the enzyme, whereas additive crystallization during fr ...19938415413
transition-state discrimination by adenosine deaminase from aspergillus oryzae.adenosine deaminase from aspergillus oryzae resembles mammalian adenosine deaminases in its ability to catalyze the hydrolytic removal of many substituents from c-6, and in the chirality at c-6 of the active isomer of the transition-state-analogue inhibitor 6-hydroxymethyl-1,6-dihydropurine ribonucleoside. the 5'-oh group of adenosine has been found to contribute a factor of 5.10(4) to transition-state stabilization by calf intestinal adenosine deaminase, and crystallographic observations sugges ...19938422418
stability of beta-galactosidase, a model protein drug, is related to water mobility as measured by 17o nuclear magnetic resonance (nmr).the inactivation of freeze-dried beta-galactosidase during storage was studied, focusing on the effect of water mobility as measured by the spin-lattice relaxation time, t1, of water using 17o nmr. inactivation of beta-galactosidase lyophilized from phosphate buffer solution was studied as a function of water content, which in turn affected the t1 of water. an increase in the water content of freeze-dried beta-galactosidase brought about an increase in the t1 of water, as well as a rise in ph. f ...19938430045
carbohydrate and aspergillus oryzae effects on intake, digestion, and milk production by dairy cows.six multiparous, ruminally cannulated holstein cows (46 dim, 584 kg of bw) and 6 primiparous holstein cows (35 dim, 506 kg of bw) were used in two 6 x 6 latin squares with 21-d periods to examine the effects of level of non-fiber carbohydrate, source of fibrous carbohydrate, and aspergillus oryzae fermentation extract on intake, digestion, and milk production. treatments were 42 and 36% non-fiber carbohydrate; shelled corn and soybean meal were replaced partially by beet pulp and dried brewers g ...19938436677
hypersensitivity pneumonitis in a soy sauce brewer caused by aspergillus oryzae. 19938436784
aspergillus nidulans nuclear proteins bind to a ccaat element and the adjacent upstream sequence in the promoter region of the starch-inducible taka-amylase a gene.aspergillus nidulans was used as an intermediate host to investigate the regulation of the taka-amylase a (taa) gene from aspergillus oryzae. the induction of taa by starch was confirmed to be regulated at the transcriptional level by analyzing the transcripts specific for taa synthesized in vitro in nuclei from starch- and glucose-grown cells. a 55 bp dna fragment containing a consensus ccaat sequence from the promoter region of the taa gene was shown to confer starch inducibility on the gene. ...19938455560
isolation and characterization of the aspergillus oryzae gene encoding aspergillopepsin o.we have cloned and determined the nucleotide sequence of a genomic dna segment from aspergillus oryzae which contains pepo, the gene encoding the aspartic proteinase, aspergillopepsin o (pepo). the organization of pepo is strikingly similar to that of pepa from a. niger var. awamori (previously called a. awamori) in that both are composed of four exons and three introns with virtually identical lengths, and the positions of the introns are exactly conserved. from the deduced amino acid (aa) sequ ...19938462873
crystals of beta-xylanase from aspergillus endo-xylanase was isolated from the culture of fungus aspergillus oryzae variant d5. the purified enzyme had a molecular weight of 24,000 and the isoelectric point of 3.6. xylanase crystals were obtained from a polyethylene glycol 6000 solution by the hanging-drop method. seeding was used for the enlargement of the crystal size. crystals belong to the monoclinic space group p2(1) with cell dimensions a = 54.9 a, b = 74.5 a, c = 50.8 a, and beta = 108.7 degrees. crystals diffract beyond 2.5 a ...19938464071
a structural domain (the lid) found in pancreatic lipases is absent in the guinea pig (phospho)lipase.typically pancreatic lipases are characterized by the following properties: (1) they are activated by lipid/water interfaces (interfacial activation), (2) they are inhibited by bile salts but reactivated by colipase (a small activator protein), and (3) they do not hydrolyze significantly phospholipids. a cdna clone encoding a guinea pig pancreatic (phospho)lipase (gpl) has been sequenced and expressed. the enzyme (recombinant as well as native) differs from other pancreatic lipases in that (1) i ...19938490016
evidence for possible involvement of an elastolytic serine protease in aspergillosis.a number of isolates of aspergillus fumigatus obtained from the hospital environment produced extracellular elastolytic activity. this activity was found to be catalyzed by a single 33-kda protein which was purified and characterized to be a serine protease. a. fumigatus, when grown on the insoluble structural material obtained from murine and bovine lung, produced the same extracellular 33-kda elastolytic protease, indicating that this enzyme is likely to be produced when the organism infects t ...19938500876
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
molecular cloning and sequence analysis of a gene encoding an aspartic proteinase from aspergillus oryzae. 19958540376
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
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
issues and advances in the use of transgenic organisms for the production of thaumatin, the intensely sweet protein from thaumatococcus danielli.the thaumatins are a class of intensely sweet proteins isolated from the fruit of the tropical plant thaumatococcus danielli. thaumatin is approved for use in many countries and has application as both a flavor enhancer and a high-intensity sweetener. the supply of naturally occurring thaumatin is limited, which has prompted extensive research into its synthesis via transgenic organisms. the gene encoding thaumatin has been introduced into various microorganisms under transcriptional control of ...19958573283
glycosidase-catalysed oligosaccharide synthesis: preparation of n-acetylchitooligosaccharides using the beta-n-acetylhexosaminidase of aspergillus oryzae.the beta-n-acetylhexosaminidase of aspergillus oryzae catalyses the formation of 2-acetamido-4-o-(2-acetamido-2-deoxy-beta-d-glucopyranosyl)-2-deoxy-d- glucopyranose (di-n-acetylchitobiose) and 2-acetamido-6-o-(2-acetamido-2-deoxy-beta-d-glucopyranosyl)-2-deoxy-d- glucopyranose from p-nitrophenyl 2-acetamido-2-deoxy-beta-d-glucopyranoside and 2-acetamido-2-deoxy-d-glucopyranose. the ratio of the two disaccharides is time-dependent. the ratio of (1-->4)- to (1-->6)-isomers is a maximum (approxima ...19958593627
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
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
characteristic expression of three amylase-encoding genes, agda, amyb, and glaa in aspergillus oryzae transformants containing multiple copies of the agda 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
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 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
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
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
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 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
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
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 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 from streptoverticillium mobaraense has been used to stabilize immobilisates produced with beta-galactosidase (beta-d-galactoside galactohydrolase, ec 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
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
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
a capillary electrophoretic study on the specificity of beta-galactosidases from aspergillus oryzae, escherichia coli, streptococcus pneumoniae, and canavalia ensiformis (jack bean).the specificities of the beta-galactosidases from aspergillus oryzae, escherichia coli, streptococcus pneumoniae, and canavalia ensiformis (jack bean) have been studied by capillary zone electrophoresis. various di- and oligosaccharides as well as a biantennary asialo n-glycan were used as substrates. following enzymatic hydrolysis, the mixtures of substrates and products were derivatized with ethyl 4-aminobenzoate and separated by high-performance capillary electrophoresis in a borate buffer sy ...19979056188
cloning, sequencing, and expression of a thermostable cellulase gene of humicola grisea.the egl2 gene coding a thermostable endoglucanase (egl2) was cloned from humicola grisea. the dna sequence of egl2 predicted two putative introns in the coding region. the deduced amino acid sequence of egl2 was 388 amino acids in length and showed 99.5% identity with the h. insolens cmc 3. in addition to tata box and caat motifs, putative crea binding sites were observed in the 5' upstream region of the egl2 gene. the egl2 gene was expressed in aspergillus oryzae, and egl2 was purified. egl2 pr ...19979058960
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
matrix-assisted laser desorption/ionization mass spectrometry (maldi) of endonuclease digests of rna.the determination of rna sequences using base- specific enzymatic cleavages is a well established method. different synthetic rna molecules were analyzed for uniformity of degradation by rnase t1, u2, a and phym under reaction conditions compatible with matrix-assisted laser desorption/ionization mass spectrometry (maldi-ms), to identify the positions of g, a and pyrimidine residues. in order to get a complete set of fragments derived from cleavage at every phosphodiester bond, the samples were ...19979115363
[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
[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
expression cloning of fungal enzyme genes; a novel approach for efficient isolation of enzyme genes of industrial relevance.expression cloning is a relatively new method for fast and efficient cloning of enzyme genes from fungi that are known to make complex enzyme mixtures. in contrast to traditional cloning methods that are usually dependent on knowledge of at least a partial amino acid sequence in order to synthesize appropriate dna probes or primers, the expression cloning method solely relies on access to reliable and sensitive enzyme assays. a representative expression cdna library is made in saccharomyces cere ...19979299700
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 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
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