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probing the anticodon loop structure in yeast trna(phe-y) with single strand-specific nuclease s1.yeast trna(phe) and trna(phe-y) are cleaved by single strand-specific endonuclease s1 at the same positions within the anticodon loop (phosphates 34, 36 and 37) and at the 3'-terminus (phosphates 75 and 76). the efficiency of the anticodon loop hydrolysis is much higher in trna(phe-y) while the cutting at the 3'-terminus is not influenced considerably by the y-base1 removal from yeast trna(phe). the effect of the y-base excision on the structure of the anticodon loop is discussed on the basis of ...19892618244
free flow electrophoresis as a method for the purification of enzymes from e. coli cell extract.the application of the four techniques of free flow electrophoresis (zone electrophoresis, isotachophoresis, isoelectric focusing and field step electrophoresis) for the purification of proteins from a complex protein mixture was investigated. for this purpose alpha-amylase (ec 3.2.1.1) from aspergillus oryzae was added and reisolated from e. coli cell extract. the chosen enzyme and the biological extract are models for many industrial separation problems. in optimized experiments purity, purifi ...19892651110
conformational stability and activity of ribonuclease t1 and mutants. gln25----lys, glu58----ala, and the double mutant.ribonuclease t1 (rnase t1) and mutants gln25----lys, glu58----ala, and the double mutant were prepared from a chemically synthesized gene, cloned and expressed in escherichia coli. the wild-type rnase t1 prepared from the cloned gene was identical in every functional and physical property examined to rnase t1 prepared from aspergillus oryzae. urea and thermal unfolding experiments show that gln25----lys is 0.9 kcal/mol more stable and glu58----ala is 0.8 kcal/mol less stable than wild-type rnase ...19892663837
studies on the accessibility of nascent non-helical peptide chains on the ribosome.the accessibility of nascent polypeptides with special structural elements to the ribosome was investigated. poly(c), poly(c, u) and poly(c, a) mrnas were translated by e. coli ribosomes in vitro. the resulting peptides which were rich in prolines, remained on the ribosomal particles or were released after addition of puromycin. a protease from aspergillus oryzae hydrolyzed the released peptides rapidly, whereas the degradation of the unreleased ones was only slightly affected. this result shows ...19892699792
characteristics of immobilized aminoacylase from aspergillus oryzae on macroporous copolymers.aminoacylase from aspergillus oryzae was adsorbed on functionallized macroporous copolymers where the enzyme showed excellent catalyzing activity and operation stability. various factors which effect the activity of the immobilized aminoacylase such as temperature, ph and ionic strength were investigated. the continuous operation of the enzyme immobilized on macroporous copolymers was compared with that of the enzyme immobilized on deae-sephadex.19902383665
effects of aspergillus oryzae fermentation extract on fermentation of amino acids, bermudagrass and starch by mixed ruminal microorganisms in vitro.the objective of this study was to examine the effects of aspergillus oryzae fermentation extract (amaferm) on the in vitro ruminal fermentation of coastal bermudagrass, soluble starch and amino acids. mixed ruminal microorganisms were incubated in anaerobic media for either 24 h (amaferm alone, soluble starch, amino acids) or 48 h (bermudagrass). amaferm was added to the incubation bottles (n = 4) at concentrations of 0, .4 or 1.0 g/liter. when mixed ruminal microorganisms were incubated with o ...19902384404
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 ...19901966694
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 ...19902079042
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 ...19902081729
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 ...19902109157
conserved residues of liquefying alpha-amylases are concentrated in the vicinity of active site.three dimensional structure of three liquefying type bacillus alpha-amylases were modeled based on sequence analyses and refined structure of aspergillus oryzae enzyme. the models suggest that the overall folding motif of alpha-amylases is conserved. the active site, substrate binding and stabilizing calcium binding residues are conserved and concentrated in a cleft between two domains. they constitute the core of alpha-amylases to which other, less conserved regions are attached. the bacterial ...19902302216
simultaneous comparison of several sequences. 19902314287
[baking ingredients, especially alpha-amylase, as occupational inhalation allergens in the baking industry].baker's asthma is the most frequent occupational lung disease in switzerland and west germany. cereal flours, and more rarely flour parasites, are implicated as the responsible allergens. based on an observation of a case of baker's asthma due to monovalent sensitization to alpha-amylase used as additive to flour, 31 bakers with occupational asthma and/or rhinitis were routinely tested by skin tests and serological rast examinations for allergic sensitivity to flour, alpha-amylase and other bake ...19902326614
influence of cultures of aspergillus oryzae on rumen and total tract digestibility of dietary components.three trials were conducted to evaluate the effect of dried cultures of aspergillus oryzae on nutrient utilization by mature holstein cows fitted with ruminal and duodenal cannulas. in trial 1, four cows (two dry and two lactating) were used to test aspergillus oryzae (3 g/d) and a control treatment at two forage amounts in a 4 x 4 latin square. trial 2 compared control, a. oryzae, and saccharomyces cerevesiae using six lactating cows in a repeated 3 x 3 latin square design. for trial 3, four la ...19902341646
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.19902143181
localization of pyruvate carboxylase in organic acid-producing aspergillus strains.the localization of pyruvate carboxylase (cytosolic or mitochondrial) was studied in nine different aspergillus species (14 strains). in some species (a. aculeatus, a. flavus, a. foetidus, a. nidulans, a. ochraceus, and a. sojae), the pyruvate carboxylase activity could be detected only in the cytosolic fraction of the cells. pyruvate carboxylase has been found only in the mitochondrial fraction of two strains of aspergillus wentii. in aspergillus oryzae and in five strains of aspergillus niger, ...19902383004
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 ...19902298207
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 ...19902110472
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 = 15.4.19902111835
hypocholesterolemic effect of the undigested fraction of soy protein. 19902126604
purification of beta-galactosidase by combined frontal and displacement chromatography. 19902113370
comparative pyrimidine- and purine-specific rnase-gold labeling on pancreatic acinar cells and isolated hepatocytes.we applied the enzyme-gold approach to investigate the potential of various ribonucleases displaying different affinities for ultrastructural localization of particular rna molecules. five specific ribonucleases were used: three from a pancreatic source, rnases a, b, and s with affinities for pyrimidine bases; and two from aspergillus oryzae, rnases t1 and t2 specific for purine bases. conditions required for preparing each rnase-gold complex, as well as for obtaining specific labelings, were de ...19901690766
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 ...19902113523
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 ...19902119802
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 ...19902120057
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 ...19902157020
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. ...19902124272
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 ...19902207069
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 ...19902229029
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 ...19902229593
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 ...19902282302
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 ...19902283420
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 ...19901369295
analysis of the conformational transitions of proteins by temperature-gradient gel electrophoresis.temperature-gradient gel electrophoresis (tgge) is a technique for studying the structural transitions of nucleic acids and proteins. a temperature gradient is formed in a horizontal slab gel perpendicular to the direction of the electric field. whereas the principle of the tgge method has previously been applied to proteins, we describe in this report the systematic optimization of tgge as a routine technique for the quantitative analysis of conformational transitions in proteins. using alpha-a ...19901706658
molecular cloning of the glucoamylase gene of aspergillus shirousami and its expression in aspergillus oryzae.the glucoamylase enzyme (gaase) gene from aspergillus shirousami was cloned and sequenced from genomic and cdna libraries. the genomic gene was located in the 5.4 kb ecori fragment. the deduced amino acid sequence of gaase contained 639 amino acid residues with a relative molecular mass of approximately 68,000 daltons (non-glycosylated form). the genomic gene of a. shirousami gaase was introduced into aspergillus oryzae. these transformants had increased gaase and raw starch degradation (rsd) ac ...19901368603
the enzymatic and molecular characteristics of saccharomycopsis alpha-amylase secreted from saccharomyces cerevisiae.the saccharomycopsis fibuligera alpha-amylase (sfamy) gene was expressed in saccharomyces cerevisiae. the highest productivity of sfamy was 70 mg per liter of culture broth. we purified sfamy from the culture broth and identified the nh2 terminal primary sequence. this sequence suggests that the sfamy gene product is synthesized as a pre-pro-precursor, and the pro-sequence is cleaved after a lys-arg sequence with the calpain-like endopeptidase encode by the kex2 gene, resulting in mature sfamy p ...19901368606
effect of diffusional resistances on the action pattern of immobilized alpha-amylase.alpha-amylase from aspergillus oryzae has been immobilized onto corn grits and porous silica (specific areas 180 and 440 m2 g-1). kinetic parameters of immobilized enzyme have been determined. immobilization of alpha-amylase results in the formation of less polymerized products resulting in an apparent decrease in the number of transglycosylation reactions, for both maltotetraose and starch as substrates, when compared with free enzyme. diffusional limitations for substrate and products have bee ...19901366409
effect of dicarboxylic acids and aspergillus oryzae fermentation extract on lactate uptake by the ruminal bacterium selenomonas ruminantium.the objective of this study was to determine the effects of l-aspartate, fumarate, l-malate, and an aspergillus oryzae fermentation extract (amaferm) on growth on lactate as well as lactate uptake by selenomonas ruminantium hd4. growth of s. ruminantium in medium that contained 2 g of dl-lactate per liter was stimulated approximately twofold by 10 mm l-aspartate, fumarate, or l-malate after 24 h. both l-aspartate and fumarate increased lactate uptake over 4-fold, while l-malate stimulated uptake ...199016348354
action pattern of alpha-amylase from aspergillus oryzae in concentrated media.the influence of concentrated maltotetraose solutions on fungal alpha-amylase activity and specificity has been determined. it has been found that the enzyme is not inhibited by 500 g/l substrate concentration and that transglycosylation reactions are increased with rising substrate concentration. the amount of oligosaccharides of polymerization degree higher than maltotetraose reaches 20% (w/w). moreover, the effect of polyhydric alcohols, known as water activity depressors has been analyzed: t ...199018592604
use of novel immobilized beta-galactosidase reactor to hydrolyze the lactose constituent of skim milk.beta-galactosidase from aspergillus oryzae immobilized in an axial-annular flow reactor was used to effect the hydrolysis of the lactose component of skim milk. nonlinear regression methods were employed to determine the kinetic parameters of four rate expressions derived from a proposed enzymatic mechanism. data taken at three different temperatures (30 degrees c, 40 degrees c, and 50 degrees c) were fit via nonlinear regression methods assuming an arrhenius temperature model for each of the pa ...199018595081
purification and some properties of soybean saponin hydrolase from aspergillus oryzae ko-2.we had investigated the enzymatic hydrolysis of soybean saponins and selected soybean saponin hydrolase from aspergillus oryzae ko-2. we attempted purification of this enzyme for further characterization. this enzyme was purified 1500-fold using ammonium sulfate fractionation and sephadex g-200 gel filtrations. the enzyme was electrophoretically homogeneous and a glycoprotein by pas staining. by gel filtration, the molecular weight of enzyme was 158,000 and sds-page showed the enzyme to have a t ...19911368672
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 ...19911930835
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 ...19911939022
performance and ruminal function development of young calves fed diets with aspergillus oryzae fermentation extract.neonatal holstein heifer (n = 72) and bull (n = 40) calves were used to study the effects of aspergillus oryzae fermentation extract (amaferm) on their performance and on rumen development. the starter diets were formulated to achieve amaferm consumption of 0, .5, 1, or 3 g per calf daily. calves were fed milk daily and allowed to consume starter and a mixture of alfalfa and bromegrass hay ad libitum. weaning was when calves consumed 550 g of starter on 2 consecutive d. weight gain and feed cons ...19911787201
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 ...19911941544
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 ...19911953634
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 ( ...19911960714
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 ...19911910319
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 ...19911913876
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. ...19911915339
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 ...19911921978
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 ...19911930833
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 ...19911930834
nucleotide sequence and expression of the glucoamylase-encoding gene (glaa) from aspergillus oryzae.the glucoamylase-encoding gene (glaa) from aspergillus oryzae was cloned using its cdna as a probe, which had been isolated previously. from comparison of nucleotide (nt) sequences of genomic clones with its cdna, the glaa gene was found to contain four short putative introns, 45-56 nt in length. the a. oryzae glaa gene shared 62% homology at the nt level with the a. niger glaa gene with the four introns located at the same position. the 5'-flanking region contained a tata box at nt-72 from the ...19911761224
proteolytic activity of two commercial proteinases from aspergillus oryzae and bacillus subtilis on ovine and bovine caseins.electrophoretic analysis of the action of two commercial enzymes, neutrase 0.5 and mkc fungal protease, on whole casein and alpha s-, beta- and kappa-caseins from cows' and ewes' milk showed that neutrase 0.5 chiefly degraded beta-casein, giving rise to peptides soluble at ph 4.6 detectable by page. in contrast, although mkc fungal protease caused intense hydrolysis of bovine beta-casein, in ovine casein it resulted in more active degradation of alpha s- than beta-casein. the latter enzyme did n ...19911765594
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 ...19911794979
genetic transfer applied to traditional sake brewing. 19911797020
clinical and immunological responses to occupational exposure to alpha-amylase in the baking industry.alpha-amylase is a starch cleaving enzyme often used in the baking industry as a flour additive. it is usually of fungal origin, produced by aspergillus oryzae. one previous report has shown ige antibodies and positive skin prick test against alpha-amylase in asthmatic bakers. this paper describes four alpha-amylase sensitised index cases with occupational asthma or rhinitis and the results of a cross sectional study of 20 workers from the same factory who were also exposed to alpha-amylase powd ...19911832939
cloning and molecular characterization of the acetamidase-encoding gene (amds) from aspergillus oryzae.we have isolated an acetamidase-encoding gene (amds) from aspergillus oryzae by heterologous hybridization using the corresponding aspergillus nidulans gene as a probe. the gene is located on a 3.5-kb saci fragment and its nucleotide (nt) sequence was determined. compared with the a. nidulans amds gene, the coding region of a. oryzae gene consists of seven exons interrupted by six introns and encodes 545 amino acid (aa) residues. the deduced aa sequence has a high degree of homology with that of ...19911840550
[isolation and properties of serine proteinase from aspergillus oryzae].a serine proteinase having an activity optimum at ph 6.7-8.2 has been isolated from amylorisine p-10x (a mixture of aspergillus oryzae enzymes) by chromatography on deae-sephadex a-50 and bacitracin sepharose 4b. the proteinase is fully inactivated by phenylmethylsulfonylfluoride and diisopropylfluorophosphonate, the specific inhibitors of the enzyme, and has a pi at ph 7.5. the molecular mass of serine proteinase is 30000 da; its amino acid composition appears as: met2, asp33, thr18, ser29, glu ...19911863668
cloning and expression in yeast of a cdna clone encoding aspergillus oryzae neutral protease ii, a unique metalloprotease.the neutral protease ii (npii) from aspergillus oryzae is a zinc-containing metalloprotease with some unique properties. to elucidate its structure, we isolated a full-length cdna clone for npii. sequence analysis reveals that npii has a prepro region consisting of 175 amino acids preceding the mature region, which consists of 177 amino acids. as compared with other microbial metalloproteases, npii is found to be unique in that it shares only a limited homology with them around two zinc ligand h ...19911886621
cloning and selective overexpression of an alkaline protease-encoding gene from aspergillus oryzae.the gene alpa encoding aspergillus oryzae alkaline protease (alp) was isolated from a genomic library of an industrial strain used in thailand by using oligodeoxyribonucleotide probes based on the published cdna sequence [tatsumi et al., agric. biol. chem. 52 (1988) 1887-1888]. the entire nucleotide sequence of the genomic clone obtained was determined. by comparison with the published cdna sequence, it was found that alp is encoded by four exons of 314, 445, 89 and 351 bp. three introns, which ...19911840548
occupational asthma in bakeries caused by sensitivity to alpha-amylase.we report on a patient with asthma induced by occupational exposure to alpha-amylase derived from aspergillus oryzae, which is a component of bread additives. a type i hypersensitivity to this enzyme was demonstrated by means of skin test, immunoassay for specific ige, and immediate bronchial provocation test response to an alpha-amylase extract.19911897689
quantitative analysis of the contribution of glu46 and asn98 to the guanosine specificity of ribonuclease 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 ...19911899029
studies on rnase t1 mutants affecting enzyme catalysis.using an escherichia coli overproducing strain secreting aspergillus oryzae rnase t1, we have constructed and characterized mutants where amino acid residues in the catalytic center have been substituted. the mutants are his40----thr, glu58----asp, glu58----gln, his92----ala and his92----phe. his92----ala and his92----phe mutants are inactive. on the basis of their kcat/km values, the mutants glu58----asp and glu58----gln show 10% and 7% residual activity, relative to wild-type rnase t1, whereas ...19911901790
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.19911904722
the glucoamylase cdna from aspergillus oryzae: its cloning, nucleotide sequence, and expression in saccharomyces cerevisiae.a cdna for aspergillus oryzae glucoamylase was cloned, using oligodeoxyribonucleotide probes derived from amino sequences of peptide fragments of the enzyme. the glucoamylase cdna, when introduced into saccharomyces cerevisiae, directed the secretion of active glucoamylase into the culture medium. the complete nucleotide sequence of the cdna contained an open reading frame encoding 612 amino acid residues. comparative studies with other fungal glucoamylases showed homologies of 67% with a. niger ...19911368680
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 ...19911368719
production and localization of enzymes on soft gel cultivation.production and localization of glutaminase and leucine aminopeptidase (lap) in soft gel cultivation were compared with those in koji and liquid cultivations. the enzymes were detected only in the whole-mycelial-mat fraction by soft gel cultivation, but in both intracellular and extracellular fractions by the other two methods. the enzyme species of glutaminase and lap in soft gel cultivation were analyzed by ion exchange and gel filtration column chromatographies. three species of glutaminase an ...19911368691
cloning and nucleotide sequences of the complementary and genomic dnas for the alkaline protease from acremonium chrysogenum.complementary dna encoding ac. chrysogenum alkaline protease (alp) was isolated from the ac. chrysogenum atcc11550 cdna library by express-blot assay. the genomic dnas encoding ac. chrysogenum alp were isolated from the ac. chrysogenum genomic dna library using the cloned cdna as a probe. the 3150 nucleotides of the gene were sequenced. the prepro-alp consists of 402 amino acids and two intervening sequences are found within the coding region. the amino acid sequence of ac. chrysogenum alp has 5 ...19911368696
isolation and characterization of the alkaline protease gene of aspergillus oryzae.the genomic dna for the alkaline protease (alp) of the fungus aspergillus oryzae was isolated using synthetic oligonucleotides as hydridization probes, and the complete nucleotide sequence was identified. the alp gene is 1374 nucleotides long and contains three introns, one of which is in the pro region and two in the mature coding region. sequences related to the tata box (tataaat) and the caat box (ccaaat) were found in the 5'-noncoding region. primer extension analysis showed that three trans ...19911368748
non-cognizable ribonucleotide, 2'amp, binds to a mutant ribonuclease t1 (y45w) at a new base-binding site but not at the guanine-recognition site.complex of a mutant ribonuclease t1 (y45w) with a non-cognizable ribonucleotide, 2'amp, has been determined and refined by x-ray diffraction at 1.7 a resolution. the 2'amp molecule locates at a new base-binding site which is remote from the guanine-recognition site, where 2'gmp was found to be bound. the nucleotide adopts the anti conformation of the glycosidic bond and c3'-exo sugar pucker. there exists a single hydrogen bond between the adenine base and the enzyme, and, therefore, the site fou ...19911655533
cloning of the nitrate-nitrite reductase gene cluster of penicillium chrysogenum and use of the niad gene as a homologous selection marker.a new homologous transformation system for the filamentous fungus penicillium chrysogenum is described. the system is based on complementation of niad mutants using the nitrate reductase structural gene (niad) of p. chrysogenum. spontaneous niad mutants were identified after selection for chlorate resistance, in growth tests and subsequent complementation with the niad gene of aspergillus oryzae. the p. chrysogenum niad gene was isolated from a genomic library using the aspergillus nidulans niad ...19911367546
an electroporation-based system for high-efficiency transformation of germinated conidia of filamentous fungi.a rapid and efficient electroporation procedure has been developed for transformation of germinating conidia of filamentous fungi. pretreatment of conidial preparations with a cell wall weakening agent, such as beta-glucuronidase, was found to be essential for successful transformation. using the qa-2+ gene of neurospora crassa, encoding the catabolic dehydroquinase, as a selectable marker with a double-mutant host strain, auxotrophic for aromatic amino acids, integration of the plasmid was obse ...19911838030
ulcerative keratomycosis--case reports on three different species of fungi.three clinical cases of fungal corneal ulcers are described to highlight the course, ocular morbidity and principles of treatment. a brief discussion of the diagnosis and management of ulcerative keratomycosis is presented.19911840452
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 ...19912072376
[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.19912064621
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 ...19912071528
comparison of the domain-level organization of starch hydrolases and related enzymes.structure-prediction and hydrophobic-cluster analysis of several starch hydrolases and related enzymes indicated the organization of eleven domain types. most enzymes possess a catalytic (beta/alpha)8-barrel and a smaller c-terminal domain as seen in crystal structures of alpha-amylase and cyclodextrin glucanotransferase. some also have a starch-granule-binding domain. enzymes breaking or forming endo-alpha-1,6 linkages contain domains n-terminal to the (beta/alpha)8-barrel.19911741756
active-site characterization of s1 nuclease. ii. involvement of histidine in catalysis.modification of the histidine residues of purified s1 nuclease resulted in loss of its single-stranded (ss)dnaase, rnaase and phosphomonoesterase activities. kinetics of inactivation indicated the involvement of a single histidine residue in the catalytic activity of the enzyme. furthermore, histidine modification was accompanied by the concomitant loss of all the activities of the enzyme, indicating the presence of a common catalytic site responsible for the hydrolysis of ssdna, rna and 3'-amp. ...19921463460
partial purification and immobilization of ribonuclease t2.a simple procedure, consisting of water extraction, heat treatment at ph 2.0, negative adsorption on deae-cellulose at ph 4.9, and concanavalin a-sepharose chromatography, was developed for the partial purification of ribonuclease (rnase) t2 from taka-diastase powder with an overall yield of 5.5%. the partially purified enzyme when coupled to aminoethyl bio-gel p-60, retained 12-16% of the activity of the soluble enzyme. temperature stability studies on rnase t2 bound to matrices, activated with ...19921418689
occupational asthma caused by alpha-amylase inhalation: clinical and immunologic findings and bronchial response patterns.inhalation of dust from different enzymes can be the cause of occupational asthma in exposed workers. alpha-amylase, derived from aspergillus oryzae, is one of these enzymes, although there are few studies in the medical literature that refer to its allergologic properties and to clinical studies in sensitized patients. the results obtained in a study performed in 83 pharmaceutical-industry workers exposed to powdered alpha-amylase are described in this article. the existence of sensitization to ...19921730831
three-dimensional structure of gln25-ribonuclease t1 at 1.84-a resolution: structural variations at the base recognition and catalytic sites.the structure of the gln25 variant of ribonuclease t1 (rnase t1) crystallized at ph 7 and at high ionic strength has been solved by molecular replacement using the coordinates of the lys25-rnase t1/2'-guanylic acid (2'gmp) complex at ph 5 [arni et al. (1988) j. biol. chem. 263, 15358-15368] and refined by energy minimization and stereochemically restrained least-squares minimization to a crystallographic r-factor of 14.4% at 1.84-a resolution. the asymmetric unit contains three molecules, and th ...19921554699
modification of subsite lys residue induced a large increase in maltosidase activity of taka-amylase a.a cross-linked modification of taka-amylase a (taa) by o-phthalaldehyde gave two enzymes, m1- and m2-taa, which had optimum ph lower than that of native taa by 0.5 to 1.0 ph units. the modified enzymes showed higher maltosidase activity, and produced glucose even at the initial period of hydrolysis, in contrast to the native taa. the modification caused more than a 500-fold decrease in the k0 value of native taa for alpha-amylase activity, but a definite increase in k0 value of 109. 1 min-1 (nat ...19921543502
cloning of the alpha-amylase cdna of aspergillus shirousamii and its expression in saccharomyces cerevisiae.alpha-amylase cdna was cloned and sequenced from aspergillus shirousamii rib2504. the putative protein deduced from the cdna open reading frame (orf) consisted of 499 amino acids with a molecular weight of 55,000. the amino acid sequence was identical to that of the orf of the taka-amylase a gene of aspergillus oryzae, while the nucleotide sequence was different at two and six positions in the cdna orf and 3' non-coding regions, respectively, so far determined. the alpha-amylase cdna was express ...19921368777
overproduction of an alpha-amylase/glucoamylase fusion protein in aspergillus oryzae using a high expression vector. 19921369066
deletion analysis of the taka-amylase a gene promoter using a homologous transformation system in aspergillus oryzae.the taka-amylase a gene (amyb) of aspergillus oryzae is induced by starch or maltose. the molecular mechanism of the induction was investigated using a fusion of the amyb promoter and the escherichia coli uida gene encoding beta-glucuronidase (gus). to identify the region responsible for high-level expression and regulation within the amyb promoter, a series of deletion promoters was constructed and introduced into the a. oryzae met locus by homologous recombination. deletion of the region betwe ...19921369079
formation of oligosaccharides from lactitol by aspergillus oryzae beta-d-galactosidase.six oligosaccharides were first formed from lactitol by a transgalactosylation reaction catalyzed by aspergillus oryzae beta-d-galactosidase. from the results of methylation analysis, ms, and 1h- and 13c-nmr studies, it was concluded that these oligosaccharides are o-beta-d-galactopyranosyl-(1----4)-o-beta-d-galactopyranosyl-(1----4)-d- glucitol, o-beta-d-galactopyranosyl-(1----3)-o-beta-d-galactopyranosyl-(1----4)-d- glucitol, o-beta-d-galactopyranosyl-(1----4)-[o-beta-d-galactopyranosyl-(1---- ...19921423346
the stereochemical course of sulphuryl transfer catalysed by arylsulphatase ii from aspergillus oryzae.phenyl [(r)-16o,17o,18o]sulphate was synthesized and used to study the stereochemical course of sulphuryl transfer to p-cresol catalysed by arylsulphatase ii from aspergillus oryzae. the reaction was shown to proceed with retention of configuration at the sulphur atom, providing evidence for the involvement of a sulpho-enzyme intermediate on the reaction pathway.19921445242
the aspergillus niger niad gene encoding nitrate reductase: upstream nucleotide and amino acid sequence comparisons.the aspergillus niger niad gene has been sequenced and the inferred nitrate reductase (nr) protein found to consist of 867 amino acid residues (97 kda). the gene is interrupted by six small introns, as deduced by comparison with the niad gene of aspergillus nidulans. the positions of these putative introns are conserved between the two fungi, although the sequences are dissimilar. the niia gene, encoding nitrite reductase, the second reductive step in the nitrate assimilation pathway, is tightly ...19921541396
hydrolysis of vicine and convicine from fababeans by microbial beta-glucosidase enzymes.the toxic glycosides vicine and convicine which are present in fababeans have been implicated in favism, an anaemic disease of humans. vicine and convicine concentrations are reduced by growth of lactobacillus plantarum on fababean suspensions. the glycosides are eliminated from the fababean substrate by the growth of the filamentous fungus fusarium graminearum. incubation of fababean suspension with concentrated culture filtrate of aspergillus oryzae, induced for extracellular beta-glucosidase ...19921644702
galactosylation at side chains of branched cyclodextrins by various beta-galactosidases.the galactosyl transfer reaction to branched cyclodextrins (cds) was investigated using lactose as a donor substrate and branched cds as acceptors by various beta-galactosidases. bacillus circulans beta-galactosidase synthesized galactosyl transfer products to branched cds, of which the galactose residues were linked at side chains of branched cds, not directly at cd rings. aspergillus oryzae and penicillium multicolor beta-galactosidases also produced derivatives galactosylated at side chains o ...19921368300
improving purification of recombinant ribonuclease t1.purification of recombinant rnase t1 and its mutants has been improved by optimizing bacterial growth conditions, periplasmic fraction preparation and the use of a precolumn. the main part of the chromatographic separation could be automated due to the reproducibility of the procedure.19921368355
a comparative study on the catalytic properties of guanyl-specific ribonucleases.the kinetic parameters of reactions catalyzed by four guanyl-specific rnases t1, pb1, th1 and sa were studied comparatively using three types of substrates; guanosine-2',3'-cyclophosphates, gpn dinucleoside phosphates and synthetic polyribonucleotides. the kinetic parameters were shown to be similar in spite of considerable differences in primary structures of these rnases, including amino acid residues of the active sites. therefore, primary structures of guanyl rnases allow for a considerable ...19921310940
functional elements of the promoter region of the aspergillus oryzae glaa gene encoding glucoamylase.analysis was made of the promoter region of the aspergillus oryzae glaa gene encoding glucoamylase. northern blots using a glucoamylase cdna as a probe indicated that the amount of mrna corresponding to the glaa gene increased when expression was induced by starch or maltose. the promoter region of the glaa gene was fused to the escherichia coli uida gene, encoding beta-glucuronidase (gus), and the resultant plasmid was introduced into a. oryzae. expression of gus protein in the a. oryzae transf ...19921339327
production of biologically active recombinant human lactoferrin in aspergillus oryzae.we report the production of recombinant human lactoferrin in aspergillus oryzae. expression of human lactoferrin (hlf), a 78 kd glycoprotein, was achieved by placing the cdna under the control of the a. oryzae alpha-amylase promoter and the 3' flanking region of the a. niger glucoamylase gene. using this system, hlf is expressed and secreted into the growth medium at levels up to 25 mg/l. the recombinant lactoferrin is indistinguishable from human milk lactoferrin with respect to its size, immun ...19921368268
on the safety of aspergillus oryzae: a review. 19921368061
controlled mycelial growth for kojic acid production using ca-alginate-immobilized fungal cells.conidia of aspergillus oryzae were immobilized in ca-alginate beads and then incubated in a nutrient medium to yield an immobilized biocatalyst producing kojic acid. the immobilized cell cultures produced kojic acid linearly during cultivation. regardless of the size of the immobilized particles, there existed an optimal nitrogen concentration for the maximum production rate of kojic acid, at which smaller bead sizes resulted in a higher production rate. when the growth of mycelia were confined ...19921368062
structural and functional analysis of the amdr regulatory gene of aspergillus oryzae.we have isolated the aspergillus oryzae homologue of the amdr regulatory gene of aspergillus nidulans by cross hybridization. sequence analysis and functional studies have shown that the amdr genes are highly conserved and functionally interchangeable between the two species. the homology between the two genes extends throughout most of the coding sequences, including sequences encoding the dna-binding domain and putative activation domains. two regions of nonconserved sequence were also identif ...19921452021
enhancement of the thermostability of the alkaline protease from aspergillus oryzae by introduction of a disulfide bond. 19921368305
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