Publications

characterization of an aspartic proteinase of mucor pusillus expressed in aspergillus oryzae.	the aspartic proteinase (mpp) gene from the zygomycete fungus mucor pusillus was introduced into an ascomycete fungus, aspergillus oryzae, by protoplast transformation using the nitrate reductase (niad) gene as the selective marker. southern blot analysis indicated that the mpp gene was integrated into the resident niad locus at a copy number of 1-2. mpp secreted by the recombinant a. oryzae was correctly processed but was more highly glycosylated than that produced in the original m. pusillus s ...	1993	8246885
the sequence and x-ray structure of the trypsin from fusarium oxysporum.	the trypsin from fusarium oxysporum is equally homologous to trypsins from streptomyces griseus, streptomyces erythraeus and to bovine trypsin. a dfp (diisopropylfluorophosphate) inhibited form of the enzyme has been crystallized from 1.4 m na2so4, buffered with citrate at ph 5.0-5.5. the crystals belong to space group p2(1) with cell parameters a = 33.43 a, b = 67.65 a, c = 39.85 a and beta = 107.6 degrees. there is one protein molecule in the asymmetric unit. x-ray diffraction data to a resolu ...	1993	8332590
preparation, properties and application of aspergillus oryzae s1 nuclease covalently bound to aminobutyl-bio-gel p-2 through its carbohydrate moiety.	purified aspergillus oryzae s1 nuclease, when covalently coupled to aminobutyl-(ab)-bio-gel p-2, via its carbohydrate moiety, retained 40-50% activity of the soluble enzyme. optimization of coupling conditions showed that the most active immobilized preparations are obtained when 50-60 units of 1 mm periodate-oxidized enzyme are allowed to react with 1 ml (packed volume) of ab-bio-gel p-2 at 4 degrees c, in the presence of 20% (v/v) ethylene glycol, for 15 h. immobilization did not change the ph ...	1993	8338642
secondary structure and temperature-induced unfolding and refolding of ribonuclease t1 in aqueous solution. a fourier transform infrared spectroscopic study.	the secondary structure of ribonuclease t1 (rnase t1) in aqueous solution and its temperature-induced structural changes have been investigated by fourier-transform infrared (ft-ir) spectroscopy. 13 to 14% alpha-helix and 32 to 33% beta-sheet were estimated from the resolution-enhanced ft-ir spectra, in agreement with the crystal structure which indicates 16% alpha-helix and 35% beta-sheet. specific ir-marker bands are assigned to the different beta-sheet structures, to the slightly bent alpha-h ...	1993	8355280
structural features of a polygalacturonase gene cloned from aspergillus oryzae kbn616.	a genomic gene encoding a polygalacturonase from aspergillus oryzae, used in soy sauce production, was cloned and sequenced. the structural gene comprises 1227 bp coding for 363 amino acids with a putative prepropeptide of 28 amino acids and the open reading frame is disrupted by two short introns of 57 bp and 81 bp. the deduced amino acid sequence of the mature protein showed 63, 63, 63 and 64% homology with those of aspergillus niger polygalacturonase i, aspergillus niger polygalacturonase ii, ...	1993	8359678
polypeptide folding of bacillus cereus atcc7064 oligo-1,6-glucosidase revealed by 3.0 a resolution x-ray analysis.	the crystal structure of an oligo-1,6-glucosidase from bacillus cereus atcc7064 was determined by the x-ray diffraction method at 3.0 a resolution. the structure was solved by the multiple isomorphous replacement method and refined to a crystallographic r-factor of 0.208, using the molecular dynamics refinement program, x-plor. the electron density map revealed the folding of a polypeptide chain consisting of 558 amino acid residues. the molecule can be subdivided into three domains (n-terminal ...	1993	8370659
effects of aspergillus oryzae fermentation extract on in vitro equine cecal fermentation.	the objective of this study was to examine the effects of aspergillus oryzae fermentation extract on the in vitro equine cecal fermentation of soluble starch, amino acids/peptides, coastal bermudagrass hay, and alfalfa hay. cecal contents were obtained from a cecally fistulated quarter horse gelding fed coastal bermudagrass and grain (70:30) either unadapted or adapted to dietary a. oryzae supplementation (2 g/d). mixed cecal microorganisms were incubated in anaerobic media for either 24 h (solu ...	1993	8376241
influence of aspergillus oryzae fermentation extract on forage intake, site of digestion, in situ degradability, and duodenal amino acid flow in steers grazing cool-season pasture.	ten ruminally and duodenally cannulated (326 +/- 28 kg) and four esophageally fistulated (394 +/- 23 kg) steers grazing cool-season pasture throughout the growing season were used to evaluate the influence of aspergillus oryzae fermentation extract (ao) supplementation on intake, forage nutrient utilization, and duodenal amino acid flow. steers grazed a predominantly smooth brome (bromus inermis l.) pasture, and measurements were taken in three periods (june, july, and august). steers were dosed ...	1993	8385089
effect of aspergillus oryzae fermentation extract (amaferm) on in vitro fiber degradation.	the influence of aspergillus oryzae fermentation extract (amaferm) on in vitro fiber degradation was determined by incubating eight ground fibrous feed-stuffs with rumen fluid and buffer inoculum. amaferm was added at 0, .4, .8, or 1.2 g/l of fermentation mixture. both ndf and adf degradabilities were determined after 96 h of incubation. addition of extract had no effect on ndf or adf degradability of pure cellulose, low endophyte fescue, wheat straw, corn silage, or prairie hay. addition of ama ...	1993	8385163
effect of nonfiber carbohydrate level and aspergillus oryzae fermentation extract on intake, digestion, and milk production in lactating dairy cows.	eight multiparous, ruminally cannulated holstein cows averaging 40 d in milk and 575 kg bw at the start of trial were in a replicated 4 x 4 latin square arrangement (28-d periods) to determine the effects of dietary nonfiber carbohydrate (nfc) level and aspergillus oryzae fermentation extract (ao) on intake, milk production, and nutrient digestibility. treatments were 42 or 35% nfc and 0 or 3 g of ao per day arranged as a 2 x 2 factorial. diets formulated to contain 21% ndf from alfalfa silage ( ...	1993	8386716
crystallization and x-ray diffraction analysis of recombinant coprinus cinereus peroxidase.	crystals suitable for an x-ray diffraction investigation have been obtained of recombinant coprinus cinereus peroxidase expressed in aspergillus oryzae. the crystals were grown by the hanging drop method with polyethylene glycol 6000 as the precipitant. a ph range from 6.2 to 8.0 and cacl2 or mgcl2 present at a concentration of 0.35 m were essential for the crystal growth. a metastable monoclinic modification can be obtained under certain conditions, and with variations in temperature they are t ...	1993	8394939
expression of aspergillus oryzae alpha-amylase gene in saccharomyces cerevisiae.	a fragment containing the full length cdna from aspergillus oryzae alpha-amylase has been amplified by pcr using specific synthetic oligonucleotides. the amplified cdna was designed to favour its expression in yeast by modifying its upstream untranslated region. it was subcloned in the expression vector pyex alpha 1, placed under the control of the yeast cyc1-gal10 promoter and used to transform saccharomyces cerevisiae. cells were then able to express and secrete active alpha-amylase to the med ...	1993	8405943
antibodies against active-site peptides common to glucosyltransferases of mutans streptococci.	polyclonal antibodies were raised against peptides derived from an active-site sequence common to the family of mutans streptococcal glucosyltransferases (gtfs). the sequence contains an aspartic acid residue that functions in formation of the enzyme transition state in catalysis. two gtfs were targeted with similar but not identical sequences in this region: one that synthesizes an alpha-1,3-linked water-insoluble glucan and a homologous gtf that synthesizes an alpha-1,6-linked water-soluble gl ...	1993	8406882
decreased protein-stabilizing effects of cryoprotectants due to crystallization.	the stabilizing effects of various additives against inactivation of an enzyme (beta-galactosidase from aspergillus oryzae) during freeze-drying were studied, with a focus on their crystallinity. the crystalline morphology of mannitol and inositol in freeze-dried cakes depended on the solute concentrations before freezing and the freeze-drying method used. the additives in their amorphous state showed concentration-dependent stabilization of the enzyme, whereas additive crystallization during fr ...	1993	8415413
transition-state discrimination by adenosine deaminase from aspergillus oryzae.	adenosine deaminase from aspergillus oryzae resembles mammalian adenosine deaminases in its ability to catalyze the hydrolytic removal of many substituents from c-6, and in the chirality at c-6 of the active isomer of the transition-state-analogue inhibitor 6-hydroxymethyl-1,6-dihydropurine ribonucleoside. the 5'-oh group of adenosine has been found to contribute a factor of 5.10(4) to transition-state stabilization by calf intestinal adenosine deaminase, and crystallographic observations sugges ...	1993	8422418
stability of beta-galactosidase, a model protein drug, is related to water mobility as measured by 17o nuclear magnetic resonance (nmr).	the inactivation of freeze-dried beta-galactosidase during storage was studied, focusing on the effect of water mobility as measured by the spin-lattice relaxation time, t1, of water using 17o nmr. inactivation of beta-galactosidase lyophilized from phosphate buffer solution was studied as a function of water content, which in turn affected the t1 of water. an increase in the water content of freeze-dried beta-galactosidase brought about an increase in the t1 of water, as well as a rise in ph. f ...	1993	8430045
carbohydrate and aspergillus oryzae effects on intake, digestion, and milk production by dairy cows.	six multiparous, ruminally cannulated holstein cows (46 dim, 584 kg of bw) and 6 primiparous holstein cows (35 dim, 506 kg of bw) were used in two 6 x 6 latin squares with 21-d periods to examine the effects of level of non-fiber carbohydrate, source of fibrous carbohydrate, and aspergillus oryzae fermentation extract on intake, digestion, and milk production. treatments were 42 and 36% non-fiber carbohydrate; shelled corn and soybean meal were replaced partially by beet pulp and dried brewers g ...	1993	8436677
hypersensitivity pneumonitis in a soy sauce brewer caused by aspergillus oryzae.		1993	8436784
aspergillus nidulans nuclear proteins bind to a ccaat element and the adjacent upstream sequence in the promoter region of the starch-inducible taka-amylase a gene.	aspergillus nidulans was used as an intermediate host to investigate the regulation of the taka-amylase a (taa) gene from aspergillus oryzae. the induction of taa by starch was confirmed to be regulated at the transcriptional level by analyzing the transcripts specific for taa synthesized in vitro in nuclei from starch- and glucose-grown cells. a 55 bp dna fragment containing a consensus ccaat sequence from the promoter region of the taa gene was shown to confer starch inducibility on the gene. ...	1993	8455560
isolation and characterization of the aspergillus oryzae gene encoding aspergillopepsin o.	we have cloned and determined the nucleotide sequence of a genomic dna segment from aspergillus oryzae which contains pepo, the gene encoding the aspartic proteinase, aspergillopepsin o (pepo). the organization of pepo is strikingly similar to that of pepa from a. niger var. awamori (previously called a. awamori) in that both are composed of four exons and three introns with virtually identical lengths, and the positions of the introns are exactly conserved. from the deduced amino acid (aa) sequ ...	1993	8462873
crystals of beta-xylanase from aspergillus oryzae.	an endo-xylanase was isolated from the culture of fungus aspergillus oryzae variant d5. the purified enzyme had a molecular weight of 24,000 and the isoelectric point of 3.6. xylanase crystals were obtained from a polyethylene glycol 6000 solution by the hanging-drop method. seeding was used for the enlargement of the crystal size. crystals belong to the monoclinic space group p2(1) with cell dimensions a = 54.9 a, b = 74.5 a, c = 50.8 a, and beta = 108.7 degrees. crystals diffract beyond 2.5 a ...	1993	8464071
a structural domain (the lid) found in pancreatic lipases is absent in the guinea pig (phospho)lipase.	typically pancreatic lipases are characterized by the following properties: (1) they are activated by lipid/water interfaces (interfacial activation), (2) they are inhibited by bile salts but reactivated by colipase (a small activator protein), and (3) they do not hydrolyze significantly phospholipids. a cdna clone encoding a guinea pig pancreatic (phospho)lipase (gpl) has been sequenced and expressed. the enzyme (recombinant as well as native) differs from other pancreatic lipases in that (1) i ...	1993	8490016
evidence for possible involvement of an elastolytic serine protease in aspergillosis.	a number of isolates of aspergillus fumigatus obtained from the hospital environment produced extracellular elastolytic activity. this activity was found to be catalyzed by a single 33-kda protein which was purified and characterized to be a serine protease. a. fumigatus, when grown on the insoluble structural material obtained from murine and bovine lung, produced the same extracellular 33-kda elastolytic protease, indicating that this enzyme is likely to be produced when the organism infects t ...	1993	8500876
studies on immobilization of aminoacylase on the functionalized polymethyl acrylate.	functionalized polymethyl acrylate was used for the immobilization of aminoacylase of aspergillus oryzae. the effects of the degree of cross-linking, the amount of pore-forming agent and their mixing rates on the activity of immobilized aminoacylase were investigated. the results showed that the activity of the immobilized aminoacylase was the highest when the carrier was a combination of 20% cross-linking and 100% pore-forming agent. the effects of substrate concentration, ionic concentration, ...	1993	8049353
on the safety of aspergillus oryzae: a review.		1992	1368061
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
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
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
enhancement of the thermostability of the alkaline protease from aspergillus oryzae by introduction of a disulfide bond.		1992	1368305
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
overproduction of an alpha-amylase/glucoamylase fusion protein in aspergillus oryzae using a high expression vector.		1992	1369066
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
effects of microorganisms on the peroxidation of lipid and fatty acid composition of fermented fish meal.	fermentation of waste fish treated with aspergillus oryzae, aspergillus sojae k, and saccharomyces cerevisiae ifo 2114 were studied independently and combined. three microorganisms decreased the pov, mda, and coy of fish meal at different rates. the optimum conditions for fermentation with the combination of three microorganisms was found at 30°c for 20-hr fermentation. almost no difference was observed in the chemical composition or amino acid spectra of the protein hydrolysates of the fermente ...	1992	27286377
cultivation characteristics of immobilized aspergillus oryzae for kojic acid production.	aspergillus oryzae in situ grown from spores entrapped in calcium alginate gel beads was used for the production of kojic acid. the immobilized cells in flask cultures produced kojic acid in a linear proportion while maintaining the stable metabolic activity for a prolonged production period. kojic acid was accumulated up to a high concentration of 83 g/l, at which the kojic acid began to crystallize, and, thus, the culture had to be replaced with fresh media for the next batch culture. the over ...	1992	18601027
three-dimensional phase-contrast imaging by a computed-tomography microscope.	we report a technique for measuring the three-dimensional variation of refractive indices in a microscopic sample. the technique is an adaptation of optical computed tomography and is effective in measuring the three-dimensional refractive-index distribution of a nonabsorbing microscopic sample. our report also includes a discussion of the conditions for the unambiguous application of the technique as well as results of experiments conducted with aspergillus oryzae (commonly known as green mold) ...	1992	20720664
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
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 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
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
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
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
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
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
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
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
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
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
genetic transfer applied to traditional sake brewing.		1991	1797020
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
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
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
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
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
[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 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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
[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
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
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
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
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
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
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
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
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