Publications
| Title | Abstract | Year Filter | PMID(sorted descending) Filter |
|---|
| identification of two essential histidine residues of ribonuclease t2 from aspergillus oryzae. | ribonuclease (rnase) t2 from aspergillus oryzae was modified by diethyl pyrocarbonate and iodoacetic acid. rnase t2 was rapidly inactivated by diethyl pyrocarbonate above ph 6.0 and by incorporation of a carboxymethyl group. no inactivation occurred in the presence of 3'amp. 1h-nmr titration and photo-chemically induced dynamic nuclear polarization experiments demonstrated that two histidine residues were involved in the active site of rnase t2. furthermore, analysis of inactive carboxymethylate ... | 1990 | 2298207 |
| effect of feeding aspergillus oryzae fermentation extract or aspergillus oryzae plus yeast culture plus mineral and vitamin supplement on performance of holstein cows during a complete lactation. | the addition of aspergillus oryzae fermentation extract (amaferm) increased milk flow and mean 3.5% fcm production during the latter stages of the full lactation trial compared with the control group and the aspergillus oryzae fermentation extract plus yeast culture plus mineral-vitamin supplement (vitaferm) group. based on the differences observed when fcm production was determined for the cows at various stages of lactation, amaferm apparently had its greatest effect during the early stages of ... | 1990 | 2283420 |
| a case of occupational allergic bronchopulmonary aspergillosis unique to japan. | a 15-year-old female was diagnosed in 1980 as having allergic bronchopulmonary aspergillosis (abpa) due to aspergillus fumigatus based on rosenberg and patterson's criteria for the disease. the patient is the eldest daughter of a family of domestic brewers of soy sauce and bean paste in a small village, an occupation unique to japan. the brewing process involved the use of aspergillus oryzae as a fermenting agent. the patient had experienced episodic wheezing and pulmonary infiltrates during the ... | 1990 | 2282302 |
| effects of feeding fungal culture extract and animal-vegetable fat on degradation of hemicellulose and on ruminal bacterial growth in heifers. | four holstein heifers cannulated in the rumen and proximal duodenum were used to analyze effects of aspergillus oryzae fermentation extract and yeast culture (amaferm micro-mix. biozyme enterprises, inc., st. joseph, mo) and 5% animal-vegetable fat on ruminal and total tract digestibilities of nutrients. heifers were assigned treatments in a 4 x 4 latin square design with a 2 x 2 factorial arrangement of treatments. few interactions between main effects were noted. feeding fat decreased ruminal ... | 1990 | 2229593 |
| primary structure of a base non-specific and adenylic acid preferential ribonuclease from aspergillus saitoi. | the complete primary structure of a base non-specific and adenylic acid preferential rnase (rnase m) from aspergillus saitoi was determined. the sequence was determined by analysis of the peptides generated by digestion of heat-denatured rnase m with lysylendopeptidase, and the peptides generated from rcm rnase m by digestion with staphylococcal v8 protease or chemical cleavage with brcn. it consisted of 238 amino acid residues and carbohydrate moiety attached to the 74th asparagine residue. the ... | 1990 | 2229029 |
| calcium binding in alpha-amylases: an x-ray diffraction study at 2.1-a resolution of two enzymes from aspergillus. | x-ray diffraction analysis (at 2.1-a resolution) of an acid alpha-amylase from aspergillus niger allowed a detailed description of the stereochemistry of the calcium-binding sites. the primary site (which is essential in maintaining proper folding around the active site) contains a tightly bound ca2+ with an unusually high number of eight ligands (o delta 1 and o delta 2 of asp175, o delta of asn121, main-chain carbonyl oxygens of glu162 and glu210, and three water molecules). a secondary bindin ... | 1990 | 2207069 |
| calculation of the relative binding free energy of 2'gmp and 2'amp to ribonuclease t1 using molecular dynamics/free energy perturbation approaches. | we present a calculation of the relative changes in binding free energy between the complex of ribonuclease t1 (rnase tr) with its inhibitor 2'-guanosine monophosphate (2'gmp) and that of rnase t1-2'-adenosine monophosphate (2'amp) by means of a thermodynamic perturbation method implemented with molecular dynamics. using the available crystal structure of the rnase t1-2'gmp complex, the structure of the rnase t1-2'amp complex was obtained as a final structure of the perturbation calculation. the ... | 1990 | 2157020 |
| asperfuran, a novel antifungal metabolite from aspergillus oryzae. | asperfuran is a novel antifungal dihydrobenzofuran derivative produced by a strain of aspergillus oryzae. asperfuran weakly inhibited chitin synthase from coprinus cinereus. this inhibition could be abolished by the addition of egg lecithin. in the agar diffusion assay asperfuran induced morphological changes in mucor miehei at very low concentrations (20 ng/disc) while growth was only partly inhibited. in hela s3 and l1210 cells it showed weak cytotoxicity, the ic50 was 25 micrograms/ml. | 1990 | 2143181 |
| hypocholesterolemic effect of the undigested fraction of soy protein. | 1990 | 2126604 | |
| crystallization and preliminary x-ray investigation of non-specific complexes of a mutant ribonuclease t1 (y45w) with 2'amp and 2'ump. | we have succeeded in crystallizing complexes of a mutant ribonuclease t1 (y45w) with the non-cognizable ribonucleotides 2'amp and 2'ump by macroscopic seeding of microcrystals of the mutant enzyme complexed with 2'gmp, which is the cognizable nucleotide inhibitor. the mutant enzyme has a tryptophan residue instead of tyr45 of the wild-type enzyme and thus this mutation enhances the binding of ribonucleotides to the enzyme. the space group is p212121 with unit cell dimensions a = 49.40 a, b = 46. ... | 1990 | 2124272 |
| primary structure of the oligo-1,6-glucosidase of bacillus cereus atcc7064 deduced from the nucleotide sequence of the cloned gene. | the gene coding for bacillus cereus atcc7064 (mesophile) oligo-1,6-glucosidase was cloned within a 2.8-kb sali-ecori fragment of dna, using the plasmid puc19 as a vector and escherichia coli c600 as a host. e. coli c600 bearing the hybrid plasmid pbce4 accumulated oligo-1,6-glucosidase in the cytoplasm. the cloned enzyme coincided absolutely with b. cereus oligo-1,6-glucosidase in its mr (65,000), in its electrophoretic behavior on a polyacrylamide gel with or without sodium dodecyl sulfate, in ... | 1990 | 2120057 |
| replacement of a cis proline simplifies the mechanism of ribonuclease t1 folding. | the refolding of ribonuclease t1 is dominated by two major slow kinetic phases that show properties of proline isomerization reactions. we report here that the molecular origin of one of these processes is the trans----cis isomerization of the ser54-pro55 peptide bond, which is cis in the native protein but predominantly trans in unfolded ribonuclease t1. this is shown by a comparison of the wild type and a designed mutant protein where ser54 and pro55 were replaced by gly54 and asn55, respectiv ... | 1990 | 2119802 |
| active-site-directed inactivation of aspergillus oryzae beta-galactosidase with beta-d-galactopyranosylmethyl-p-nitrophenyltriazene. | beta-d-galactopyranosylmethyl-p-nitrophenyltriazene (beta-galmnt), a specific inhibitor of beta-galactosidase, was isolated as crystals by hplc and its chemical and physicochemical characteristics were examined. aspergillus oryzae beta-galactosidase was inactivated by the compound. we studied the inhibition mechanism in detail. the inhibitor was hydrolyzed by the enzyme to p-nitroaniline and an active intermediate (beta-galactopyranosylmethyl carbonium or beta-galactopyranosylmethyldiazonium), w ... | 1990 | 2113523 |
| purification of beta-galactosidase by combined frontal and displacement chromatography. | 1990 | 2113370 | |
| purification of recombinant ribonuclease t1 expressed in escherichia coli. | a protocol for the rapid purification of ribonuclease t1 expressed from a chemically synthesized gene cloned into escherichia coli is described. qae ion-exchange and sephadex g-50 chromatography are used to give over 300 mg (88% yield) of pure ribonuclease t1 from 61 of liquid culture in 3 days. we also report a new absorption coefficient for rnase t1: e1%278 nm = 15.4. | 1990 | 2111835 |
| ph dependence of the urea and guanidine hydrochloride denaturation of ribonuclease a and ribonuclease t1. | to investigate the ph dependence of the conformational stability of ribonucleases a and t1, urea and guanidine hydrochloride denaturation curves have been determined over the ph range 2-10. the maximum conformational stability of both proteins is about 9 kcal/mol and occurs near ph 4.5 for ribonuclease t1 and between ph 7 and 9 for ribonuclease a. the ph dependence suggests that electrostatic interactions among the charged groups make a relatively small contribution to the conformational stabili ... | 1990 | 2110472 |
| closely related glycosylation patterns of recombinant human il-2 expressed in a cho cell line and natural il-2. | we report here the study of the glycosylation pattern of human recombinant (r) il2 expressed in a chinese hamster ovary (cho) cell line. the human ril2 secreted by this high-producing recombinant cho cell line was metabolically radiolabelled with [35s]-methionine, or with [3h]-glucosamine and [3h]-galactose, purified to homogeneity, and then characterized. the electrophoretic analysis of the [35s]-methionine-labelled proteins present in the culture medium of the cho cell line showed that the ril ... | 1990 | 2109157 |
| crystallographic characterization of wild-type and mutant ribonuclease t1 complexes with several ribonucleotides. | ribonuclease t1 and the mutant enzymes were cocrystallized with several ribonucleotides, including non-hydrolyzable substrate analogs of di- and triribonucleotides, which have a novel guanylate in which the 2'-hydroxyl group of the ribose is replaced by a fluorine atom. one of the mutant enzymes has a tryptophan residue, instead of tyr45 of the wild-type enzyme, to enhance the binding of ribonucleotides to the enzyme and the other mutant enzyme has histidine and aspartate residues, instead of as ... | 1990 | 2081729 |
| purification of s1 nuclease from aspergillus oryzae by recycling isoelectric focusing. | we describe the purification of a single-strand nuclease from aspergillus oryzae using the first commercial prototype of an instrument (rf3tm) designed by milan bier et al. for preparative-scale isoelectric focusing. protein separation takes place entirely in solution, with shear-stabilized laminar flow counteracting convective disturbances generated by the electric field. conditions for isoelectric focusing were determined by focusing fractions with nuclease activity, following chromatography o ... | 1990 | 2079042 |
| isolation and characterisation of an extracellular alkaline protease of aspergillus fumigatus. | aspergillus fumigatus secreted an inducible alkaline protease (alpase) when cultivated in the presence of collagen (200 micrograms/ml) as sole nitrogen and carbon source. proteolytic activity was maximum at ph 9.0 with azocollagen as substrate. the enzyme, which was the major protein found in the supernate of a liquid culture, was purified by ammonium sulphate precipitation and gel filtration. the mr was determined to be 33 kda by gel filtration and sodium dodecyl sulphate-polyacrylamide gel ele ... | 1991 | 2072376 |
| influence of feeding aspergillus oryzae fermentation extract on the milk yields, eating patterns, and body temperatures of lactating cows. | trials were conducted to evaluate effects of a fermentation extract of aspergillus oryzae (ao) on milk production and composition, diet digestibility, and rectal temperature changes in lactating dairy cows. treatments were incorporated as a top dressing at the morning feeding and consisted of control (90 g/d of ground sorghum) or ao (3 g of culture + 87 g of ground sorghum daily). twenty-four mid-lactation holstein cows were paired for production in lactation trial 1 (lt-1). in lactation trial 2 ... | 1991 | 2071528 |
| [structure and antitumor activity of polysaccharides from the micelles of aspergillus oryzae]. | three polysaccharide fractions active against mammary gland adenocarcinoma ca-755 in mice but inactive against sarcoma c-180 were isolated from aspergillus oryzae strain 5214. the main fraction, extracted from the mycelium by cold dilute alkali in the presence of sodium borohydride, was shown to be linear (1----3)-a-d-glucopyranan (pseudonigeran) according to 13c nmr spectroscopy, partial acid hydrolysis and periodate oxidation data. | 1991 | 2064621 |
| calorimetric analysis of microbial growth: with special reference to quantitative evaluation of drug action. | our research method for the calorimetric characterization of the biological effects of drugs and other chemicals on metabolic activities of living cells is outlined. the effects of various substances on different microbial systems were studied quantitatively using a calorimeter, and the results were used to plot drug potency curves for each drug. the method was also used to study microbial activity in soil. it was found to be a useful technique for the quantitative characterization of pollutants ... | 1990 | 1966694 |
| in defence of aldehyde-osmium fixation and critical-point drying for characterization of seed-storage fungi by scanning electron microscopy. | low-temperature scanning electron microscopy cannot be used as a general approach when resolution of definitive surface features is required for fungal characterization, because of the production of a pervading exudate by some of the species, strains or varieties. as the resolution of the light microscope is obviously limiting, scanning electron microscopy remains the best microscopical mode. results are presented for four species within the aspergillus flavus group which indicate that minimal ( ... | 1991 | 1960714 |
| aluminofluorides and beryllofluorides as inhibitors of sulphatases. analogues of hydrogen sulphate? | the inhibition of fluoride of sulphatase a from ox liver and of the sulphatases of helix pomatia and aspergillus oryzae is decreased by edta and increased by al3+ or be2+, implicating aluminofluorides and beryllofluorides in the reaction. the inhibition, which is reversible, takes several minutes to develop fully and, at least for the sulphatase of h. pomatia, is of a non-linear mixed competitive-non-competitive type. it is suggested that the aluminofluorides and beryllofluorides are acting as a ... | 1991 | 1953634 |
| evidence of steric factors in the fungitoxic mechanisms of 8-quinolinol and its 5- and 7-halogenated analogues. | antifungal studies were made of mixtures of minimal inhibitory concentrations (mics) of 8-quinolinol and its 5- and 7-halo analogues against six fungi: aspergillus niger, a. oryzae, trichoderma viride, myrothecium verrucaria, mucor cirinelloides, and trichophyton mentagrophytes. mixtures of 8-quinolinol with 5- or 7-fluoro-8-quinolinol and of 5- and 7-fluoro-8-quinolinol showed additive activity, and their respective toxicities were reversed by l-cysteine. these results suggested a common mechan ... | 1991 | 1941544 |
| amino acid sequence of nuclease s1 from aspergillus oryzae. | the amino acid sequence of nuclease s1, a nuclease which cleaves both single-stranded dna and rna, from aspergillus oryzae was determined. reduced and s-carboxymethylated or s-aminoethylated nuclease s1 was digested with achromobacter protease i, staphylococcus aureus v8 protease, or endoproteinase asp-n. peptides thus obtained were purified by reverse-phase high-performance liquid chromatography and sequenced, and the complete primary structure was established. nuclease s1 consists of a single ... | 1991 | 1939022 |
| structure and molecular model refinement of aspergillus oryzae (taka) alpha-amylase: an application of the simulated-annealing method. | monoclinic crystals of a neutral alpha-amylase from aspergillus oryzae, containing three molecules in the asymmetric unit, have been reported previously and studied at 3 a resolution [matsuura, kunusoki, harada & kakudo (1984). j. biochem. 95, 697-702]. here we report the solution of the structure of this enzyme in a different crystal form (space group p2(1)2(1)2(1), a = 50.9, b = 67.2, c = 132.7 a), with only one molecule in the asymmetric unit. the structure was solved by the molecular replace ... | 1991 | 1930835 |
| solution of the structure of aspergillus niger acid alpha-amylase by combined molecular replacement and multiple isomorphous replacement methods. | the crystal structure of aspergillus niger acid alpha-amylase was solved by a combination of multiple isomorphous replacement and molecular replacement methods. the atomic coordinates of aspergillus oryzae (taka) alpha-amylase (entry 2taa in the protein data bank) and experimental diffraction data from a new monoclinic crystal form of taka alpha-amylase, were used during the procedure. sequence identity between the two proteins is approximately 80%. the atomic parameters derived from the molecul ... | 1991 | 1930834 |
| x-ray analysis of cubic crystals of the complex formed between ribonuclease t1 and guanosine-3',5'-bisphosphate. | the complex formed between ribonuclease t1 (rnase t1) and guanosine-3',5'-bisphosphate (3',5'-pgp) crystallizes in the cubic space group i23 with alpha = 86.47 (4) a. x-ray data were collected on a four-circle diffractometer to 3.2 a resolution and the structure was determined by molecular-replacement methods [ultima; rabinovich & shakked (1984). acta cryst. a40, 195-200] based on the rnase t1 coordinates taken from the complex with guanosine-2'-phosphate. refinement converged at 16.6% for 1540 ... | 1991 | 1930833 |
| construction of a fusion gene comprising the taka-amylase a promoter and the escherichia coli beta-glucuronidase gene and analysis of its expression in aspergillus oryzae. | northern blot analysis of glucose-grown and starch-grown mycelia of aspergillus oryzae rib40 was conducted using the cloned taka-amylase a (taa) gene as a probe. the amount of mrna homologous to the taa gene was increased when this fungus was grown with starch as a sole carbon source. in order to analyze the induction mechanism, we inserted the escherichia coli uida gene encoding beta-glucuronidase (gus) down-stream of the taa promoter and introduced the resultant fusion gene into the a. oryzae ... | 1991 | 1921978 |
| primary structure of nuclease p1 from penicillium citrinum. | the primary structure of nuclease p1, which cleaves both rna and single-stranded dna, from penicillium citrinum was elucidated. the complete amino acid sequence consisting of 270 residues was determined by analysis of peptides obtained by digestion with achromobacter protease i of the reduced and s-aminoethylated protein and by digestion with staphylococcus aureus v8 protease of the reduced and s-carboxymethylated protein. four half-cystine residues were assigned to cys72-cys217 and cys80-cys85. ... | 1991 | 1915339 |
| cloning and nucleotide sequence of the genomic ribonuclease t2 gene (rntb) from aspergillus oryzae. | using synthetic oligonucleotide probes, we have cloned a genomic dna sequence encoding a ribonuclease (rnase) t2 gene (rntb) from aspergillus oryzae on a 4.8 kb hindiii fragment. dna sequence analysis of the rnase t2 revealed the following: (1) the gene is arranged as five exons and four introns; (2) the deduced amino acid sequence contains 239 amino acid residues of the mature enzyme. in addition, there exist 17 amino acid residues thought to be a signal peptide sequence at the n-terminus and 2 ... | 1991 | 1913876 |
| affinity labeling of a subsite of taka-amylase a by the fluorescent reagent o-phthalaldehyde. | a cross-linked modification of lys residue located at the subsite of the enzyme active site of taka-amylase a was attained by the use of the fluorescent reagent of o-phthalaldehyde (opa). the fluorescence and uv absorption at 337 nm derived from the isoindole ring, which was produced by cross-linking through the epsilon-amino group of lys and the thiol group of the cys residue, provided the evidence for the opa-mediated inactivation of taka-amylase a. kinetic analysis showed that 1 mol of opa pe ... | 1991 | 1910319 |
| specificity of guanylic rnases to polynucleotide substrates. | kinetic parameters kcat and km were measured for cleavage of poly i, poly a, poly u, poly c and poly i poly c by guanyl-specific rnases sa, pb1 and t1 and compared with that of guanyl-preferential rnase bi. catalytic efficiencies of the investigated enzymes to polynucleotide substrates vary considerably. the structural basis for specificity of these rnases is discussed. a hypothesis is suggested that ser-56 plays an important role in recognition of poly a by rnase bi. | 1991 | 1904722 |
| 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 ... | 1991 | 1901790 |
| quantitative analysis of the contribution of glu46 and asn98 to the guanosine specificity of ribonuclease t1. | in the crystal structure of the ribonuclease t1 (rnase t1; ec 3.1.27.3)-2'-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 ... | 1991 | 1899029 |
| 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. | 1991 | 1897689 |
| 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 ... | 1991 | 1886621 |
| [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 ... | 1991 | 1863668 |
| 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 ... | 1991 | 1840550 |
| 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 ... | 1991 | 1840548 |
| 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. | 1991 | 1840452 |
| 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 ... | 1991 | 1838030 |
| 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 ... | 1991 | 1832939 |
| genetic transfer applied to traditional sake brewing. | 1991 | 1797020 | |
| isolation, characterization, and primary structure of a base non-specific and adenylic acid preferential ribonuclease with higher specific activity from trichoderma viride. | in 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 ... | 1991 | 1794979 |
| 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 ... | 1991 | 1787201 |
| 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 ... | 1991 | 1765594 |
| 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 ... | 1991 | 1761224 |
| 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. | 1991 | 1741756 |
| 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 ... | 1992 | 1730831 |
| 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 ... | 1990 | 1706658 |
| 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 ... | 1990 | 1690766 |
| 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 ... | 1991 | 1655533 |
| 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 ... | 1992 | 1644702 |
| 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 ... | 1992 | 1554699 |
| 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 ... | 1992 | 1543502 |
| 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 ... | 1992 | 1541396 |
| structure of ribonuclease t1 complexed with zinc(ii) at 1.8 a resolution: a zn2+.6h2o.carboxylate clathrate. | in 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 ... | 1992 | 1515106 |
| 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 ... | 1992 | 1500571 |
| 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 ... | 1992 | 1500561 |
| 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 ... | 1992 | 1476369 |
| 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. ... | 1992 | 1463460 |
| 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 ... | 1992 | 1457048 |
| 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 ... | 1992 | 1452021 |
| 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. | 1992 | 1445242 |
| 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---- ... | 1992 | 1423346 |
| 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 ... | 1992 | 1418689 |
| secretion of aspergillus oryzae alkaline protease in an osmophilic yeast, zygosaccharomyces rouxii. | to 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 ... | 1990 | 1369295 |
| 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 ... | 1992 | 1369079 |
| overproduction of an alpha-amylase/glucoamylase fusion protein in aspergillus oryzae using a high expression vector. | 1992 | 1369066 | |
| 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 ... | 1992 | 1368777 |
| 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 ... | 1991 | 1368748 |
| efficient production of taka-amylase a by trichoderma viride. | an 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 ... | 1991 | 1368719 |
| 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 ... | 1991 | 1368696 |
| 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 ... | 1991 | 1368691 |
| 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 ... | 1991 | 1368680 |
| 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 ... | 1991 | 1368672 |
| 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 ... | 1990 | 1368606 |
| 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 ... | 1990 | 1368603 |
| 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. | 1992 | 1368355 |
| enhancement of the thermostability of the alkaline protease from aspergillus oryzae by introduction of a disulfide bond. | 1992 | 1368305 | |
| 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 ... | 1992 | 1368300 |
| 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 ... | 1992 | 1368268 |
| 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 ... | 1992 | 1368062 |
| on the safety of aspergillus oryzae: a review. | 1992 | 1368061 | |
| 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 ... | 1991 | 1367546 |
| 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 ... | 1990 | 1366409 |
| 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 ... | 1992 | 1339327 |
| 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 ... | 1992 | 1310940 |
| 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 ... | 1992 | 1307441 |
| 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 ... | 1992 | 1281415 |
| 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 ... | 1992 | 1280919 |
| 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 ... | 1976 | 1278151 |
| 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. | 1976 | 1273594 |
| 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 ... | 1976 | 1272803 |
| 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 ... | 1976 | 1262346 |
| 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 ... | 1976 | 1261542 |
| 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 ... | 1976 | 1253642 |