[production of immobilized alpha-amylase and its properties].active immobilized alpha-amylase was obtained when applying ae-cellulose chromatography and glutaric dialdehyde. the time of the enzyme interaction with the carrier, amount of glutaric dialdehyde necessary for binding, optimal enzyme: carrier ratio as well as the methods for desiccation of the immobilized amylase preparations were specified. conditions are selected for alpha-amylase stabilization with the presence of glutaric aldehyde. immobilized amylase as compared to free enzyme is shown to b ...1976982618
fermentative production of limit dextrinase by aspergillus oryzae. 1976992794
aspergillus oryzae (nrrl strain 1988): a clarification. 1976996551
subsite mapping of enzymes. application of the depolymerase computer model to two the preceding paper (allen and thoma, 1976) we developed a depolymerase computer model, which uses a minimization routine to establish a subsite map for a depolymerase. in the present paper we show how the model is applied to experimental data for two alpha-amylases. michaelis parameters and bond-cleavage frequencies for substrates of chain lengths up to twelve glucosyl units have been reported for bacillus amyloliquefaciens, and a subsite map has been proposed for this enzyme [thoma et al. ( ...1976999630
conformation of mammalian 5.8 s ribosomal rna: s1 nuclease as a probe. 19761001452
effect of brinase (protease i from aspergillus oryzae) on the elimination of fibrin from the lungs and pulmonary damage in rats with induced intravascular coagulation and inhibited fibrinolysis. 19761006629
structure of cerebroside in aspergillus oryzae.structural studies on cerebroside isolated from aspergillus oryzae were carried out using mainly gas-liquid chromatography-mass spectrometry. the major component fatty acids were 2-hydroxystearic and 2-hydroxy-trans-octadecenoic acid; branched 17?-methyl nonadecasphingadienine isomers were the predominant long-chain bases. the component sugar was only glucose. the structure of the major cerebrosides was assumed to be n-2'-hydroxystearoyl-1-o-glucosyl-17?-methyl sphingadienine and n-2'-hydroxyoct ...19761009131
proteinase inhibitors from cereal grains. 19761012027
[hydrolysis of insoluble bone collagen by crystalline alpha-amylase from aspergillus oryzae].the paper deals with hydrolysis of bone collagen thrice recrystallized alpha-amylase. the enzyme action was estimated by the amount of released glycopeptides, free carbohydrates, alpha-amine groups and total nitrogen in the soluble part of the hydrolyzate. the protease admixture in the alpha-amylase preparation was found by means of the aspergillus oryzae protease. the data obtained testify to the fact that hydrolysis of collagen under the effect of crystalline alpha-amylase occurs only due to t ...19761021920
thrombolytic treatment with i.v. brinase of advanced arterial obliterative disease of the limbs.the material includes 17 patients suffering from different degrees of chronic peripheral arterial disease (11 chronic patients stage iii and iv, two patients with acute arterial occlusion, and four patients stage ii). presence and extent of arterial occlusion was ascertained by initial arteriography. in twelve of the patients amputation had been considered. the patients were treated by a series of i.v. infusions of brinase, a proteolytic enzyme from aspergillus oryzae. the brinase inhibitor capa ...19751053595
comparison of the effects of linear and cyclic phosphates on the adsorption of proteins by human enamel.treatment of human enamel powder with cyclic trimetaphosphate or linear tripolyphosphate changes subsequent adsorption of amylase, lysozyme, and salivary protein from aqueous solutions. treated enamel samples adsorb more lysozyme, less amylase, and more salivary protein (at concentrations exceeding 12 mug/ml) than controls treated with distilled water.19751056353
transglycosylation by the exo-type of beta-n-acetyglucosaminidase from human parotid saliva. 19751057392
the enhancing influence of proteolysis on e rosette forming lymphocytes (t cells) in vivo and in vitro.the t lymphocytes populations of 22 young healthy adults, 21 healthy middle aged and older blood donors, 35 non-pregnant women of child bearing age and 14 patients with advanced malignant disease were assessed and compared. it was found that the mean t cell counts in the middle aged and older controls were significantly lower than in the healthy young adults and were further reduced in the patients with malignant disease. the addition of the proteolytic agent brinase (protease 1 obtained from as ...19751080669
investigation into the use of aspergillus oryzae s1 nuclease in the presence of solvents which destabilize or prevent dna secondary structure: formaldehyde, formamide, and glyoxal. 19751093442
fatal aspergillosis in imported parrots.spontaneous fatal aspergillosis occurred in several species of parrots imported from latin america, australia, malaya and ghana for studies on the control of psittacosis. over a period of 4 years, 655 parrots were imported for use in these studies. all birds that died during these investigations were necropsied, and the internal organs of 45 were found to have macroscopic lesions suggestive of aspergillosis. of these 45 suspected cases, 32 were confirmed as aspergillosis by both histopathology a ...19751097931
reversal of fungitoxicity of 8-quinolinols and their copper (ii) bischelates. i. amino acids and related compounds.the effect of amino acids and related compounds on the toxicity of 8-quinolinols and their copper (ii) bischelates to aspergillus oryzae (atcc 1011) was studied. none of the compounds tested except the thiol-containing compounds, cysteine, cysteamine, glutathione, and n-acetylcysteine reversed the inhibitory action of 8-quinolinol but not that of 5-iodo-8-quinolinol or bis (8-quinolinolato) copper (ii). it appears that the mechanism(s) of fungitoxicity of 8-quinolinol is different from that of 5 ...19751116048
specific hydrolysis of the cohesive ends of bacteriophage lambda dna by three single strand-specific nucleases.procedures have been worked out for aspergillus nuclease s1 and mung been nuclease to quantitatively cleave off both of the 12-nucleotide long, single-stranded cohesive ends of lambdadna. this cleavage is indicated by the almost complete elimination of the repair incorporation of radioactive nucleotides by dna polymerase into the digested dna. with s1 nuclease, cleavage was complete at 10 degrees as well as at 30 degrees. under the conditions for quantitative cleavage of the single-stranded regi ...19751141222
transformation of malathion by a fungus, aspergillus oryzae, isolated from a freshwater pond. 19751148416
[action of leucine aminopeptidase 1 and 2 (aspergillus oryzae 410) on various synthetic peptides (author's transl)].by use of di- or tripeptides as substrates, lap 1 and lap 2, 2 fractions from aspergillus oryzae hydrolyze oligopeptides that possess leucine as n-terminal amino acid. lap 1 fraction also hydrolyzes the histidyl bond. both fractions have no activity towards peptides as glutathion, gly-pro-ala; they have low or no activity towards tyrosyl and tryptophanyl bond.19751157844
triacetonamine formation in fungal extracts. 19751159172
in vitro recognition of carcinogen-induced local denaturation sites native dna by s1 endonuclease from aspergillus oryzae. 19751161018
purification and some properties of the soluble and membrane-bound adenosine deaminases of micrococcus sodonensis atcc 11880 and their distribution within the family micrococcus sodonensis and some other micrococcus species, adenosien deaminase is present both as a membran-bound and a soluble enzyme; the membran-bound adenosine deaminase can be extracted with n-butanol, and may account for up to 5% of the total cellular adenosine deaminase activity. in a number oc comparative tests, no differences between the two enzyme forms could be found, thus they are believed to be similar molecular species; the purified membran-bound or soluble enzyme had a molecula ...19751168529
[growth patterns of fungi]. 19751169575
a chemical and biological assessment of aspergillus oryzae and other filamentous fungi as protein sources for simple stomached animals. 19751172167
s1 nuclease hydrolysis of single-stranded nucleic acids with partial double-stranded configuration.the single-strand specific nuclease s1 from aspergillus oryzae (ec was purified 600-fold in 16% yield from dried mycelia. determination of the isoelectric point of s1 nuclease as 4.3-4.4 allowed adjustment of chromatographic conditions such that the enzyme was isolated free of contaminating ribonucleases t1 and t2. s1 nuclease so purified was used for removal of single-stranded portions from the rna of the escherichia coli phage ms2, which has a helical content of about 65% in vitro. a ...19751182098
studies on the arylsulphatase and phenol sulphotransferase activities of aspergillus oryzae.1. crude extracts of aspergillus oryzae grown under conditions of sulphur limitation possess high arylsulphatase activity. 2. this activity can be greatly enhanced by the inclusion of tyramine or a number of other phenols in the assay medium. 3. the arylsulphatase activity of these extracts can be resolved into three distinct fractions by chromatography on deae-cellulose. 4. the effect of tyramine is restricted to one of these fractions only. 5. evidence is presented which indicates that this ef ...19751200998
the atmospheric fungal flora of the athens metropolitan the research programme of the department of microbiology of the athens university the nature of the mycological flora of the athenian air was studied. the research took place during the calendar year 1971. the open air was sampled twice weekly from two observation stations. the open plate technique was used, petri dishes containing sabourand's agar being exposed for 15 minutes. a total of 180 plates were exposed, and 1714 fungal colonies were isolated; these were subcultured and identified as ...19751207717
[denaturation stabilization of alpha-amylase from aspergillus oryzae].denaturation of alpha-amylase from aspergillus oryzae was studied under the effect of heating urea and some other denaturating agents. inhibition in the enzyme denaturation, deviation from the first order equation and, consequently, establishment of the false equilibrium in the system are shown. the values are calculated for the reaction rate constants of alpha-amylase denaturation under the effect to heat (40 degrees c) and urea. a method is developed for isolating native amylase stabilized by ...19751209773
[the amino acid sequence of the double-headed protein proteinase inhibitor from dog submandibular glands, i. structural homology to the pancreatic secretory trypsin inhibitors (author's transl)].the primary structure of the broad specificity proteinase inhibitor from dog submandibular glands was elucidated. the inhibitor consists of a single polypeptide chain of 117 amino acids which is folded into two domains (heads) connected by a peptide of three amino acid residues. both domains i and ii show a clear structural homology to each other as well as to the single-headed pancreatic secretory trypsin inhibitors (kazal type). the trypsin reactive site (-cys-pro-arg-leu-his-glx-pro-ile-cys-) ...19751213678
[effect of iaa on the content of rna in the cells and isolated nuclei of aspergillus oryzae].the content of rna in mycelium and isolated nuclei of aspergillus oryzae 3-9-15 was studied after its growth during two days on the modified capek medium containing 0.01 and 10 mg% of iaa. a direct correlation has been established between the extracellular concentration of iaa and its content in the mycelium and isolated nuclei of the fungus. the stimulating effect of the extracellular iaa on the content of rna in the isolated nuclei was optimal after one hour of incubation in the capek medium c ...19751214614
[chymotrypsin-like proteinases in microbial proteolytic complexes].a considerable amount of proteinase of chemotrypsin type (ao-ct) was found in the proteolytic system of asp. oryzae, strain "h"-476. the proteinase produces an intensive effect when hydrolyzing p-nitrophenyl acetate (p-npa) -- 18-20 times as strong as that of crystalline chemotrypsin. comparison of the ao-ct and chemotrypsin properties resulted in proving the enzyme nature of the p-npa hydrolysis reaction. a study of ao-ct specificity with the presence of such substrates as p-npa, o-nitro-phenyl ...19751216345
studies on the substrate specificity of taka-amylase a. xii. investigation of the active site of taka-amylase a by examining the properties of p-phenylazobenzoyl taka-amylase a.1. when p-phenylazobenzoyl taka-amylase a (phab-taa) was incubated at ph 6.5 with hydroxylamine for 3 hr at 20degrees, some of the p-phenylazobenzoyl residues that had been introduced into taka-amylase a (taa) [1, 4-alpha-d-glucan glucanohydrolase, ec, aspergillus oryzae] were liberated as a hydroxamic acid, and the activity pattern of phab-taa changed to that of intact taa. this result suggested that the p-phenylazobenzoyl residues liberated had been bound to the tyrosyl residue located ...19751225912
chemical mutagenesis. ii. some observations on the new morphogenetic variations induced in aspergillus oryzae wehmer by 6-azauracil. 19751242247
protein and amino acid changes in peanut (arachis hypogaea l.) seeds infected with aspergillus oryzae. 19761245677
renaturation of bacteriophage lambda dna. determination of the optimal renaturation conditions using a single-strand-specific dnase and alkaline-sucrose-gradient assay system.reannealed hybrid molecules of wild-type bacteriophage lambda dna were prepared in aqueous solutions of formamide at a variety of nacl concentrations at both room temperature ( 22 degrees c) and 37 degrees c. treatment of the hybrid dna molecules with the single-strand-specific nuclease s1 from aspergillus oryzae followed by alkaline sucrose gradient sedimentation was used to monitor the extent and fidelity of hybridization. the optimal renaturation conditions at room temperature were found to b ...19761248479
the in situ formation of dna-dna duplexes of drosophila virilis satellite dna.the dna of drosophila virilis brains and imaginal discs was labeled in vitro to a specific activity of 6 x 10(-5) dpm/mug, using an organ culture medium. the dna was fractionated on neutral and alkaline csc1 gradients and the heavy strands of satellite i annealed in situ to denatured polytene chromosomes from squash preparations of larval salivary glands. nuclease s1 from aspergillus oryzae was used to digest the unpaired ssdna, resulting in a distinct labeling of the alpha-heterochromatin in th ...19761253642
nuclease s1 cleavage and the primary structure of mitochondrial dna.the single-strand-specific nuclease s1 from aspergillus oryzae rapidly converts superhelical mitochondrial dna (african green monkey cells, vero atcc; ccl 81) into nicked circular dna. these nicked mitochondrial dna molecules contain two nicks, one in each strand. the phosphodiester backbones are cleaved during this reaction at or near sites that are alkali-labile. in a second slow reaction the circular mitochondrial dna is converted into a linear duplex dna. permutation tests indicate that this ...19761261542
purification and properties of the nuclease inhibitor of aspergillus oryzae and kinetics of its interaction with crystalline nuclease o.a nuclease inhibitor found in the mycelia of aspergillus oryzae has been purified 158,000-fold by ammonium sulfate precipitation, chromatography on sephadex g-75, deae-sephadex a-50 and bio-gel p-60 columns, preparative disc electrophoresis on acrylamide gel, and electrofocusing in ampholite. the purified inhibitor is nearly homogeneous as judged by disc electrophoresis. it shows a typical ultraviolet absorption curve for protein, and the inhibitory activity is inactivated by chymotrypsin. the i ...19761262346
sepcific fragmentation of dna heteroduplex molecules of two bacteriophage lambda mutants with endonuclease si from aspergillus oryzae.heteroduplex dna molecules of two bacteriophage mutants (lambda b2 and lambda i434ct68) were obtained by the method of molecular hybridization. these heteroduplexes possessed two types of loops formed as a result of: a) deletion in one of the dna strands; and b) substitution of a dna fragment for nonhomological one. the digestion of heteroduplexes with single-stranded specific nuclease si from aspergillus oryzae produced two fragments at 37 degrees c and three ones at 55 degrees c. the separatio ...19761272803
aflatoxin production by a variant of aspergillus oryzae (nrrl strain 1988) on cowpeas (vigna sinensis).aflatoxin b1, b2, g1, and g2 are produced when a variant of aspergillus oryzae (nrrl strain 1988) is grown on cowpeas or rice. the present study indicates that a strain of aspergillus oryzae approved for use in food processing is variable and the resulting variant, unlike the parent strain, has a propensity to produce aflatoxin.19761273594
the structure of kinetoplast dna. 2. characterization of a novel component of high complexity present in the kinetoplast dna network of crithidia luciliae.1. degradation of highly purified kinetoplast dna (kdna) networks with restriction endonucleases yields "extra" bands in agarose gels that are absent from digests of mini-circles. each of the five endonucleases tested, i.e. alui, hapii, ecori, hsu and hindii + iii, yields a unique set of "extra" bands. the "extra" bands consist of linear dna; they are not mini-circle oligomers and their added molecular weight, calculated from mobility in gels, are around 2 x 10(7). double digests with two restri ...19761278151
the zymogram method for detection of ribonucleases after isoelectric focusing: analysis of multiple forms of human, bovine, and microbial enzymes.a zymogram method for detection of in situ ribonuclease (rnase) activity, combined with isoelectric focusing in a thin layer of polyacrylamide gel (ief-page), has been developed. after incubation with a dried agarose film containing substrate rna, ethidium bromide, and an appropriate reaction buffer, which was placed tightly on the top of the focused gel, sharp and distinct dark bands corresponding to rnase isoenzymes on a fluorescent background appeared under uv light. addition of urea to the i ...19921280919
high level expression of the synthetic human lysozyme gene in aspergillus oryzae.aspergillus oryzae was transformed with a synthetic gene consisting of a chicken lysozyme signal sequence and a mature human lysozyme (hly) sequence. the transformants secreted active hly (about 1.2 mg/l) when the hly gene was expressed under the control of the taka-amylase a gene (amyb) promoter. western blot analysis suggested that the secreted protein was immunoreactive with anti-human lysozyme antibody and the signal peptide was correctly cleavaged off in the a. oryzae transformants. the tra ...19921281415
distribution of aflatoxin-producing moulds and aflatoxins in dairy cattle feed and raw milk.distribution of aflatoxigenic moulds and aflatoxin b1 in yugoslav dairy cattle feeds as well as the presence of aflatoxin b1 and m1 in raw milk, was tested. the experiments were carried out through three years (in all seasons). samples were taken from state and private farms in vojvodina. feeds were contaminated in 83-100% with moulds. fungi of farms in vojvodina. feeds were contaminated in 83-100% with moulds. fungi of aspergillus flavus-oryzae group were present permanently and the highest inc ...19921307441
a comparative study on the catalytic properties of guanyl-specific ribonucleases.the kinetic parameters of reactions catalyzed by four guanyl-specific rnases t1, pb1, th1 and sa were studied comparatively using three types of substrates; guanosine-2',3'-cyclophosphates, gpn dinucleoside phosphates and synthetic polyribonucleotides. the kinetic parameters were shown to be similar in spite of considerable differences in primary structures of these rnases, including amino acid residues of the active sites. therefore, primary structures of guanyl rnases allow for a considerable ...19921310940
functional elements of the promoter region of the aspergillus oryzae glaa gene encoding glucoamylase.analysis was made of the promoter region of the aspergillus oryzae glaa gene encoding glucoamylase. northern blots using a glucoamylase cdna as a probe indicated that the amount of mrna corresponding to the glaa gene increased when expression was induced by starch or maltose. the promoter region of the glaa gene was fused to the escherichia coli uida gene, encoding beta-glucuronidase (gus), and the resultant plasmid was introduced into a. oryzae. expression of gus protein in the a. oryzae transf ...19921339327
effect of diffusional resistances on the action pattern of immobilized alpha-amylase.alpha-amylase from aspergillus oryzae has been immobilized onto corn grits and porous silica (specific areas 180 and 440 m2 g-1). kinetic parameters of immobilized enzyme have been determined. immobilization of alpha-amylase results in the formation of less polymerized products resulting in an apparent decrease in the number of transglycosylation reactions, for both maltotetraose and starch as substrates, when compared with free enzyme. diffusional limitations for substrate and products have bee ...19901366409
cloning of the nitrate-nitrite reductase gene cluster of penicillium chrysogenum and use of the niad gene as a homologous selection marker.a new homologous transformation system for the filamentous fungus penicillium chrysogenum is described. the system is based on complementation of niad mutants using the nitrate reductase structural gene (niad) of p. chrysogenum. spontaneous niad mutants were identified after selection for chlorate resistance, in growth tests and subsequent complementation with the niad gene of aspergillus oryzae. the p. chrysogenum niad gene was isolated from a genomic library using the aspergillus nidulans niad ...19911367546
on the safety of aspergillus oryzae: a review. 19921368061
controlled mycelial growth for kojic acid production using ca-alginate-immobilized fungal cells.conidia of aspergillus oryzae were immobilized in ca-alginate beads and then incubated in a nutrient medium to yield an immobilized biocatalyst producing kojic acid. the immobilized cell cultures produced kojic acid linearly during cultivation. regardless of the size of the immobilized particles, there existed an optimal nitrogen concentration for the maximum production rate of kojic acid, at which smaller bead sizes resulted in a higher production rate. when the growth of mycelia were confined ...19921368062
production of biologically active recombinant human lactoferrin in aspergillus oryzae.we report the production of recombinant human lactoferrin in aspergillus oryzae. expression of human lactoferrin (hlf), a 78 kd glycoprotein, was achieved by placing the cdna under the control of the a. oryzae alpha-amylase promoter and the 3' flanking region of the a. niger glucoamylase gene. using this system, hlf is expressed and secreted into the growth medium at levels up to 25 mg/l. the recombinant lactoferrin is indistinguishable from human milk lactoferrin with respect to its size, immun ...19921368268
galactosylation at side chains of branched cyclodextrins by various beta-galactosidases.the galactosyl transfer reaction to branched cyclodextrins (cds) was investigated using lactose as a donor substrate and branched cds as acceptors by various beta-galactosidases. bacillus circulans beta-galactosidase synthesized galactosyl transfer products to branched cds, of which the galactose residues were linked at side chains of branched cds, not directly at cd rings. aspergillus oryzae and penicillium multicolor beta-galactosidases also produced derivatives galactosylated at side chains o ...19921368300
enhancement of the thermostability of the alkaline protease from aspergillus oryzae by introduction of a disulfide bond. 19921368305
improving purification of recombinant ribonuclease t1.purification of recombinant rnase t1 and its mutants has been improved by optimizing bacterial growth conditions, periplasmic fraction preparation and the use of a precolumn. the main part of the chromatographic separation could be automated due to the reproducibility of the procedure.19921368355
molecular cloning of the glucoamylase gene of aspergillus shirousami and its expression in aspergillus oryzae.the glucoamylase enzyme (gaase) gene from aspergillus shirousami was cloned and sequenced from genomic and cdna libraries. the genomic gene was located in the 5.4 kb ecori fragment. the deduced amino acid sequence of gaase contained 639 amino acid residues with a relative molecular mass of approximately 68,000 daltons (non-glycosylated form). the genomic gene of a. shirousami gaase was introduced into aspergillus oryzae. these transformants had increased gaase and raw starch degradation (rsd) ac ...19901368603
the enzymatic and molecular characteristics of saccharomycopsis alpha-amylase secreted from saccharomyces cerevisiae.the saccharomycopsis fibuligera alpha-amylase (sfamy) gene was expressed in saccharomyces cerevisiae. the highest productivity of sfamy was 70 mg per liter of culture broth. we purified sfamy from the culture broth and identified the nh2 terminal primary sequence. this sequence suggests that the sfamy gene product is synthesized as a pre-pro-precursor, and the pro-sequence is cleaved after a lys-arg sequence with the calpain-like endopeptidase encode by the kex2 gene, resulting in mature sfamy p ...19901368606
purification and some properties of soybean saponin hydrolase from aspergillus oryzae ko-2.we had investigated the enzymatic hydrolysis of soybean saponins and selected soybean saponin hydrolase from aspergillus oryzae ko-2. we attempted purification of this enzyme for further characterization. this enzyme was purified 1500-fold using ammonium sulfate fractionation and sephadex g-200 gel filtrations. the enzyme was electrophoretically homogeneous and a glycoprotein by pas staining. by gel filtration, the molecular weight of enzyme was 158,000 and sds-page showed the enzyme to have a t ...19911368672
the glucoamylase cdna from aspergillus oryzae: its cloning, nucleotide sequence, and expression in saccharomyces cerevisiae.a cdna for aspergillus oryzae glucoamylase was cloned, using oligodeoxyribonucleotide probes derived from amino sequences of peptide fragments of the enzyme. the glucoamylase cdna, when introduced into saccharomyces cerevisiae, directed the secretion of active glucoamylase into the culture medium. the complete nucleotide sequence of the cdna contained an open reading frame encoding 612 amino acid residues. comparative studies with other fungal glucoamylases showed homologies of 67% with a. niger ...19911368680
production and localization of enzymes on soft gel cultivation.production and localization of glutaminase and leucine aminopeptidase (lap) in soft gel cultivation were compared with those in koji and liquid cultivations. the enzymes were detected only in the whole-mycelial-mat fraction by soft gel cultivation, but in both intracellular and extracellular fractions by the other two methods. the enzyme species of glutaminase and lap in soft gel cultivation were analyzed by ion exchange and gel filtration column chromatographies. three species of glutaminase an ...19911368691
cloning and nucleotide sequences of the complementary and genomic dnas for the alkaline protease from acremonium chrysogenum.complementary dna encoding ac. chrysogenum alkaline protease (alp) was isolated from the ac. chrysogenum atcc11550 cdna library by express-blot assay. the genomic dnas encoding ac. chrysogenum alp were isolated from the ac. chrysogenum genomic dna library using the cloned cdna as a probe. the 3150 nucleotides of the gene were sequenced. the prepro-alp consists of 402 amino acids and two intervening sequences are found within the coding region. the amino acid sequence of ac. chrysogenum alp has 5 ...19911368696
efficient production of taka-amylase a by trichoderma efficient heterologous protein production system was developed in trichoderma viride, a very efficient cellulase producer. an expression vector containing the taka-amylase a gene from aspergillus oryzae, which was fused to the strong promoter and signal peptide sequence of the cellobiohydrolase 1 gene (cbh1) of t. viride, and the hygromycin b resistance gene was used to transform protoplasts of t. viride. using hygromycin b resistance, a frequency of 3 transformants per microgram dna on avera ...19911368719
isolation and characterization of the alkaline protease gene of aspergillus oryzae.the genomic dna for the alkaline protease (alp) of the fungus aspergillus oryzae was isolated using synthetic oligonucleotides as hydridization probes, and the complete nucleotide sequence was identified. the alp gene is 1374 nucleotides long and contains three introns, one of which is in the pro region and two in the mature coding region. sequences related to the tata box (tataaat) and the caat box (ccaaat) were found in the 5'-noncoding region. primer extension analysis showed that three trans ...19911368748
cloning of the alpha-amylase cdna of aspergillus shirousamii and its expression in saccharomyces cerevisiae.alpha-amylase cdna was cloned and sequenced from aspergillus shirousamii rib2504. the putative protein deduced from the cdna open reading frame (orf) consisted of 499 amino acids with a molecular weight of 55,000. the amino acid sequence was identical to that of the orf of the taka-amylase a gene of aspergillus oryzae, while the nucleotide sequence was different at two and six positions in the cdna orf and 3' non-coding regions, respectively, so far determined. the alpha-amylase cdna was express ...19921368777
overproduction of an alpha-amylase/glucoamylase fusion protein in aspergillus oryzae using a high expression vector. 19921369066
deletion analysis of the taka-amylase a gene promoter using a homologous transformation system in aspergillus oryzae.the taka-amylase a gene (amyb) of aspergillus oryzae is induced by starch or maltose. the molecular mechanism of the induction was investigated using a fusion of the amyb promoter and the escherichia coli uida gene encoding beta-glucuronidase (gus). to identify the region responsible for high-level expression and regulation within the amyb promoter, a series of deletion promoters was constructed and introduced into the a. oryzae met locus by homologous recombination. deletion of the region betwe ...19921369079
secretion of aspergillus oryzae alkaline protease in an osmophilic yeast, zygosaccharomyces produce aspergillus oryzae alkaline protease (alp) in an osmophilic yeast zygosaccharomyces rouxii, we constructed an expression plasmid consisting of the z. rouxii glyceraldehyde-3-phosphate dehydrogenase (gapdh) promoter, the prepro-alp cdna of a. oryzae, the whole sequence of z. rouxii plasmid psr1, and the g418 resistant gene. the resulting plasmid, when introduced into z. rouxii cells, directed the secretion of a large amount (about 300 mg/l) of alp into the culture medium. the n-terminu ...19901369295
partial purification and immobilization of ribonuclease t2.a simple procedure, consisting of water extraction, heat treatment at ph 2.0, negative adsorption on deae-cellulose at ph 4.9, and concanavalin a-sepharose chromatography, was developed for the partial purification of ribonuclease (rnase) t2 from taka-diastase powder with an overall yield of 5.5%. the partially purified enzyme when coupled to aminoethyl bio-gel p-60, retained 12-16% of the activity of the soluble enzyme. temperature stability studies on rnase t2 bound to matrices, activated with ...19921418689
formation of oligosaccharides from lactitol by aspergillus oryzae beta-d-galactosidase.six oligosaccharides were first formed from lactitol by a transgalactosylation reaction catalyzed by aspergillus oryzae beta-d-galactosidase. from the results of methylation analysis, ms, and 1h- and 13c-nmr studies, it was concluded that these oligosaccharides are o-beta-d-galactopyranosyl-(1----4)-o-beta-d-galactopyranosyl-(1----4)-d- glucitol, o-beta-d-galactopyranosyl-(1----3)-o-beta-d-galactopyranosyl-(1----4)-d- glucitol, o-beta-d-galactopyranosyl-(1----4)-[o-beta-d-galactopyranosyl-(1---- ...19921423346
the stereochemical course of sulphuryl transfer catalysed by arylsulphatase ii from aspergillus oryzae.phenyl [(r)-16o,17o,18o]sulphate was synthesized and used to study the stereochemical course of sulphuryl transfer to p-cresol catalysed by arylsulphatase ii from aspergillus oryzae. the reaction was shown to proceed with retention of configuration at the sulphur atom, providing evidence for the involvement of a sulpho-enzyme intermediate on the reaction pathway.19921445242
structural and functional analysis of the amdr regulatory gene of aspergillus oryzae.we have isolated the aspergillus oryzae homologue of the amdr regulatory gene of aspergillus nidulans by cross hybridization. sequence analysis and functional studies have shown that the amdr genes are highly conserved and functionally interchangeable between the two species. the homology between the two genes extends throughout most of the coding sequences, including sequences encoding the dna-binding domain and putative activation domains. two regions of nonconserved sequence were also identif ...19921452021
immobilization of aminoacylase from aspergillus oryzae on synthetic modified polyacrylamides.a series of acrylamide-bisacrylamide copolymers modified by the mannich reaction was prepared. the immobilization of aminoacylase from aspergillus oryzae on the copolymers was studied. all the polymers adsorbed the enzyme and the activity of the immobilized enzyme dependent on the amine used, viz. secondary amine, diamine, or aniline derivative. however, the activity was also influenced by the degree of crosslinking of the polymer. the surface morphology of the dimethylamine-modified polymer, wi ...19921457048
active-site characterization of s1 nuclease. ii. involvement of histidine in catalysis.modification of the histidine residues of purified s1 nuclease resulted in loss of its single-stranded (ss)dnaase, rnaase and phosphomonoesterase activities. kinetics of inactivation indicated the involvement of a single histidine residue in the catalytic activity of the enzyme. furthermore, histidine modification was accompanied by the concomitant loss of all the activities of the enzyme, indicating the presence of a common catalytic site responsible for the hydrolysis of ssdna, rna and 3'-amp. ...19921463460
operational stability of enzymes. acylase-catalyzed resolution of n-acetyl amino acids to enantiomerically pure l-amino acids.the method of measuring enzyme deactivation by monitoring necessary addition of fresh enzyme to keep a constant degree of conversion in a cstr at constant [e] x tau, the product of concentration of active enzyme [e] and residence time tau, was successfully applied to acylase i from porcine kidney and aspergillus oryzae fungus. fungal enzyme was found to be more stable than kidney enzyme. activation by both co2+ and zn2+ ions also yielded increased operational enzyme stability: co2+ and zn2+ are ...19921476369
influence of feeding varying levels of amaferm on performance of lactating dairy cows.amaferm, a fermentation extract of aspergillus oryzae, was fed as a top-dressing to dairy cows at 0, 1.5, 3, and 6 g/d in two lactation trials using 64 cows in 1989. lactation trial 1 was conducted in the spring (march to may) and used 40 cows averaging 75 dim for a 70-d treatment period. lactation trial 2 was during the summer (june to july). twenty-four cows averaging 140 dim were employed in a 60-d study. measurements included milk yield, feed intake, bw, rectal temperatures, respiration rate ...19921500561
effect of direct-fed microbials on rumen microbial fermentation.nonbacterial, direct-fed microbials added to ruminant diets generally consist of aspergillus oryzae fermentation extract, or saccharomyces cerevisiae cultures, or both. results from in vivo research have been variable regarding effects of direct-fed microbials on ruminant feedstuff utilization and performance. some research has shown increased weight gains, milk production, and total tract digestibility of feed components, but others have shown little influence of direct-fed microbials on these ...19921500571
structure of ribonuclease t1 complexed with zinc(ii) at 1.8 a resolution: a zn2+.6h2o.carboxylate order to study the inhibitory effect of zn2+ on ribonuclease t1 [rnase t1; itaya & inoue (1982). biochem. j. 207, 357-362], the enzyme was cocrystallized with 2 mm zn2+, ph 5.2, from a solution containing 55% (v/v) 2-methyl-2,4-pentanediol. the crystals are orthorhombic, p2(1)2(1)2(1), a = 48.71 (1), b = 46.51 (1), c = 41.14 (1) a, z = 4, v = 93203 a3. the crystal structure was determined by molecular replacement and refined by restrained least-squares methods based on fhkl for 8291 unique re ...19921515106
the aspergillus niger niad gene encoding nitrate reductase: upstream nucleotide and amino acid sequence comparisons.the aspergillus niger niad gene has been sequenced and the inferred nitrate reductase (nr) protein found to consist of 867 amino acid residues (97 kda). the gene is interrupted by six small introns, as deduced by comparison with the niad gene of aspergillus nidulans. the positions of these putative introns are conserved between the two fungi, although the sequences are dissimilar. the niia gene, encoding nitrite reductase, the second reductive step in the nitrate assimilation pathway, is tightly ...19921541396
modification of subsite lys residue induced a large increase in maltosidase activity of taka-amylase a.a cross-linked modification of taka-amylase a (taa) by o-phthalaldehyde gave two enzymes, m1- and m2-taa, which had optimum ph lower than that of native taa by 0.5 to 1.0 ph units. the modified enzymes showed higher maltosidase activity, and produced glucose even at the initial period of hydrolysis, in contrast to the native taa. the modification caused more than a 500-fold decrease in the k0 value of native taa for alpha-amylase activity, but a definite increase in k0 value of 109. 1 min-1 (nat ...19921543502
three-dimensional structure of gln25-ribonuclease t1 at 1.84-a resolution: structural variations at the base recognition and catalytic sites.the structure of the gln25 variant of ribonuclease t1 (rnase t1) crystallized at ph 7 and at high ionic strength has been solved by molecular replacement using the coordinates of the lys25-rnase t1/2'-guanylic acid (2'gmp) complex at ph 5 [arni et al. (1988) j. biol. chem. 263, 15358-15368] and refined by energy minimization and stereochemically restrained least-squares minimization to a crystallographic r-factor of 14.4% at 1.84-a resolution. the asymmetric unit contains three molecules, and th ...19921554699
hydrolysis of vicine and convicine from fababeans by microbial beta-glucosidase enzymes.the toxic glycosides vicine and convicine which are present in fababeans have been implicated in favism, an anaemic disease of humans. vicine and convicine concentrations are reduced by growth of lactobacillus plantarum on fababean suspensions. the glycosides are eliminated from the fababean substrate by the growth of the filamentous fungus fusarium graminearum. incubation of fababean suspension with concentrated culture filtrate of aspergillus oryzae, induced for extracellular beta-glucosidase ...19921644702
non-cognizable ribonucleotide, 2'amp, binds to a mutant ribonuclease t1 (y45w) at a new base-binding site but not at the guanine-recognition site.complex of a mutant ribonuclease t1 (y45w) with a non-cognizable ribonucleotide, 2'amp, has been determined and refined by x-ray diffraction at 1.7 a resolution. the 2'amp molecule locates at a new base-binding site which is remote from the guanine-recognition site, where 2'gmp was found to be bound. the nucleotide adopts the anti conformation of the glycosidic bond and c3'-exo sugar pucker. there exists a single hydrogen bond between the adenine base and the enzyme, and, therefore, the site fou ...19911655533
comparative pyrimidine- and purine-specific rnase-gold labeling on pancreatic acinar cells and isolated hepatocytes.we applied the enzyme-gold approach to investigate the potential of various ribonucleases displaying different affinities for ultrastructural localization of particular rna molecules. five specific ribonucleases were used: three from a pancreatic source, rnases a, b, and s with affinities for pyrimidine bases; and two from aspergillus oryzae, rnases t1 and t2 specific for purine bases. conditions required for preparing each rnase-gold complex, as well as for obtaining specific labelings, were de ...19901690766
analysis of the conformational transitions of proteins by temperature-gradient gel electrophoresis.temperature-gradient gel electrophoresis (tgge) is a technique for studying the structural transitions of nucleic acids and proteins. a temperature gradient is formed in a horizontal slab gel perpendicular to the direction of the electric field. whereas the principle of the tgge method has previously been applied to proteins, we describe in this report the systematic optimization of tgge as a routine technique for the quantitative analysis of conformational transitions in proteins. using alpha-a ...19901706658
occupational asthma caused by alpha-amylase inhalation: clinical and immunologic findings and bronchial response patterns.inhalation of dust from different enzymes can be the cause of occupational asthma in exposed workers. alpha-amylase, derived from aspergillus oryzae, is one of these enzymes, although there are few studies in the medical literature that refer to its allergologic properties and to clinical studies in sensitized patients. the results obtained in a study performed in 83 pharmaceutical-industry workers exposed to powdered alpha-amylase are described in this article. the existence of sensitization to ...19921730831
comparison of the domain-level organization of starch hydrolases and related enzymes.structure-prediction and hydrophobic-cluster analysis of several starch hydrolases and related enzymes indicated the organization of eleven domain types. most enzymes possess a catalytic (beta/alpha)8-barrel and a smaller c-terminal domain as seen in crystal structures of alpha-amylase and cyclodextrin glucanotransferase. some also have a starch-granule-binding domain. enzymes breaking or forming endo-alpha-1,6 linkages contain domains n-terminal to the (beta/alpha)8-barrel.19911741756
nucleotide sequence and expression of the glucoamylase-encoding gene (glaa) from aspergillus oryzae.the glucoamylase-encoding gene (glaa) from aspergillus oryzae was cloned using its cdna as a probe, which had been isolated previously. from comparison of nucleotide (nt) sequences of genomic clones with its cdna, the glaa gene was found to contain four short putative introns, 45-56 nt in length. the a. oryzae glaa gene shared 62% homology at the nt level with the a. niger glaa gene with the four introns located at the same position. the 5'-flanking region contained a tata box at nt-72 from the ...19911761224
proteolytic activity of two commercial proteinases from aspergillus oryzae and bacillus subtilis on ovine and bovine caseins.electrophoretic analysis of the action of two commercial enzymes, neutrase 0.5 and mkc fungal protease, on whole casein and alpha s-, beta- and kappa-caseins from cows' and ewes' milk showed that neutrase 0.5 chiefly degraded beta-casein, giving rise to peptides soluble at ph 4.6 detectable by page. in contrast, although mkc fungal protease caused intense hydrolysis of bovine beta-casein, in ovine casein it resulted in more active degradation of alpha s- than beta-casein. the latter enzyme did n ...19911765594
performance and ruminal function development of young calves fed diets with aspergillus oryzae fermentation extract.neonatal holstein heifer (n = 72) and bull (n = 40) calves were used to study the effects of aspergillus oryzae fermentation extract (amaferm) on their performance and on rumen development. the starter diets were formulated to achieve amaferm consumption of 0, .5, 1, or 3 g per calf daily. calves were fed milk daily and allowed to consume starter and a mixture of alfalfa and bromegrass hay ad libitum. weaning was when calves consumed 550 g of starter on 2 consecutive d. weight gain and feed cons ...19911787201
isolation, characterization, and primary structure of a base non-specific and adenylic acid preferential ribonuclease with higher specific activity from trichoderma order to elucidate the structure-function relationship of rnases belonging to the rnase t2 family (base non-specific and adenylic acid-preferential rnase), an rnase of this family was purified from trichoderma viride (rnase trv) to give three closely adjacent bands with rnase activity on slab-gel electrophoresis in a yield of 20%. the three rnases gave single band with the same mobility on slab-gel electrophoresis after endoglycosidase f digestion. the enzymatic properties including base spec ...19911794979
genetic transfer applied to traditional sake brewing. 19911797020
clinical and immunological responses to occupational exposure to alpha-amylase in the baking industry.alpha-amylase is a starch cleaving enzyme often used in the baking industry as a flour additive. it is usually of fungal origin, produced by aspergillus oryzae. one previous report has shown ige antibodies and positive skin prick test against alpha-amylase in asthmatic bakers. this paper describes four alpha-amylase sensitised index cases with occupational asthma or rhinitis and the results of a cross sectional study of 20 workers from the same factory who were also exposed to alpha-amylase powd ...19911832939
an electroporation-based system for high-efficiency transformation of germinated conidia of filamentous fungi.a rapid and efficient electroporation procedure has been developed for transformation of germinating conidia of filamentous fungi. pretreatment of conidial preparations with a cell wall weakening agent, such as beta-glucuronidase, was found to be essential for successful transformation. using the qa-2+ gene of neurospora crassa, encoding the catabolic dehydroquinase, as a selectable marker with a double-mutant host strain, auxotrophic for aromatic amino acids, integration of the plasmid was obse ...19911838030
ulcerative keratomycosis--case reports on three different species of fungi.three clinical cases of fungal corneal ulcers are described to highlight the course, ocular morbidity and principles of treatment. a brief discussion of the diagnosis and management of ulcerative keratomycosis is presented.19911840452
cloning and selective overexpression of an alkaline protease-encoding gene from aspergillus oryzae.the gene alpa encoding aspergillus oryzae alkaline protease (alp) was isolated from a genomic library of an industrial strain used in thailand by using oligodeoxyribonucleotide probes based on the published cdna sequence [tatsumi et al., agric. biol. chem. 52 (1988) 1887-1888]. the entire nucleotide sequence of the genomic clone obtained was determined. by comparison with the published cdna sequence, it was found that alp is encoded by four exons of 314, 445, 89 and 351 bp. three introns, which ...19911840548
cloning and molecular characterization of the acetamidase-encoding gene (amds) from aspergillus oryzae.we have isolated an acetamidase-encoding gene (amds) from aspergillus oryzae by heterologous hybridization using the corresponding aspergillus nidulans gene as a probe. the gene is located on a 3.5-kb saci fragment and its nucleotide (nt) sequence was determined. compared with the a. nidulans amds gene, the coding region of a. oryzae gene consists of seven exons interrupted by six introns and encodes 545 amino acid (aa) residues. the deduced aa sequence has a high degree of homology with that of ...19911840550
[isolation and properties of serine proteinase from aspergillus oryzae].a serine proteinase having an activity optimum at ph 6.7-8.2 has been isolated from amylorisine p-10x (a mixture of aspergillus oryzae enzymes) by chromatography on deae-sephadex a-50 and bacitracin sepharose 4b. the proteinase is fully inactivated by phenylmethylsulfonylfluoride and diisopropylfluorophosphonate, the specific inhibitors of the enzyme, and has a pi at ph 7.5. the molecular mass of serine proteinase is 30000 da; its amino acid composition appears as: met2, asp33, thr18, ser29, glu ...19911863668
cloning and expression in yeast of a cdna clone encoding aspergillus oryzae neutral protease ii, a unique metalloprotease.the neutral protease ii (npii) from aspergillus oryzae is a zinc-containing metalloprotease with some unique properties. to elucidate its structure, we isolated a full-length cdna clone for npii. sequence analysis reveals that npii has a prepro region consisting of 175 amino acids preceding the mature region, which consists of 177 amino acids. as compared with other microbial metalloproteases, npii is found to be unique in that it shares only a limited homology with them around two zinc ligand h ...19911886621
occupational asthma in bakeries caused by sensitivity to alpha-amylase.we report on a patient with asthma induced by occupational exposure to alpha-amylase derived from aspergillus oryzae, which is a component of bread additives. a type i hypersensitivity to this enzyme was demonstrated by means of skin test, immunoassay for specific ige, and immediate bronchial provocation test response to an alpha-amylase extract.19911897689
quantitative analysis of the contribution of glu46 and asn98 to the guanosine specificity of ribonuclease the crystal structure of the ribonuclease t1 (rnase t1; ec'-gmp complex the hydrogen-bonding potential of the guanine base is saturated [arni, r., heinemann, u., tokuoka, r., & saenger, w. (1988) j. biol. chem. 263, 15358-15368]. the oxygens of the glu46 carboxylate and the asn98 main-chain carbonyl act as hydrogen-bond acceptors for the n(1)h-c(2)-n(2)h2 part of the base. we measured the transesterification kinetics of wild-type and glu46ala rnase t1 using the gpu, ipu, and xpu s ...19911899029
studies on rnase t1 mutants affecting enzyme catalysis.using an escherichia coli overproducing strain secreting aspergillus oryzae rnase t1, we have constructed and characterized mutants where amino acid residues in the catalytic center have been substituted. the mutants are his40----thr, glu58----asp, glu58----gln, his92----ala and his92----phe. his92----ala and his92----phe mutants are inactive. on the basis of their kcat/km values, the mutants glu58----asp and glu58----gln show 10% and 7% residual activity, relative to wild-type rnase t1, whereas ...19911901790
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