TitleAbstractYear(sorted ascending)
[isolation of a specific inhibitor of microbial serine proteinase from kidney bean seeds].a protein acting as a specific inhibitor of microbial serine proteinases was isolated from kidney bean seeds. the purification procedure included complex formation between the inhibitor and aspergillus oryzae proteinase. the protein with a mr approximately 10 000 inhibits subtilisin and asp. oryzae proteinase but does not affect trypsin and chymotrypsin. the inhibitor molecule contains no half-cystine residues.19836357292
effect of drugs on the activity of fungal limit dextrinase & alpha-amylase. 19836360862
stereochemical course of dna hydrolysis by nuclease s1.nuclease s1 hydrolyzes the sp-diastereomer of 5'-o-(2'-deoxyadenosyl)-3'-o-thymidyl phosphorothioate in h2(18)o to [18o]deoxyadenosine 5'-o-phosphorothioate which can be phosphorylated enzymatically to the sp-diastereomer of [alpha-18o]deoxyadenosine 5'-o-(1-thiotriphosphate). 31p nmr spectroscopy shows the oxygen-18 in this compound to be in a nonbridging position at the alpha-phosphorus, indicating that the hydrolysis reaction catalyzed by nuclease s1 proceeds with inversion of configuration a ...19836296110
[effect of the physiological conditions on alpha-amylase and glucoamylase formation by a selected strain of aspergillus oryzae].the production of alpha-amylase and glucoamylase by a selected strain of aspergillus oryzae was investigated using different carbon and nitrogen sources. the best and most economic fermentation medium for the production of the both amylases in submerged cultures had the following composition (in %): defatted rice brain, 8; corn steep liquor, 3; mgso4 x 7h2o, 0.1; kh2po4, 0.1; cacl2, 0.1. the optimum ph was 5.0. the optimal conditions for biosynthesis of the amylases were as follows: cultivation ...19836413831
effects of thiol protease inhibitors on intracellular degradation of exogenous beta-galactosidase in cultured human skin fibroblasts.the effects of low molecular weight (lmw) protease inhibitors of microbial origin were evaluated on the intracellular degradation of beta-galactosidase purified from aspergillus oryzae and taken up by cultured human skin fibroblasts with beta-galactosidase deficiency. only thiol protease inhibitors showed an effect to increase the enzyme activity. e-64, a specific inhibitor of thiol proteases, prolonged 3-fold a half life of the exogenous beta-galactosidase and when the enzyme was supplied as li ...19836414834
regulation of the oxidative phase of the pentose phosphate cycle in aspergillus oryzae (ahlburg). i. induction of glucose-6-phosphate dehydrogenase.the activity of glucose-6-phosphate dehydrogenase changes when the mycelium is grown on different carbon sources. in the presence of ribose, the activity of the enzyme is approximately 3 times higher than when the mycelium is grow in a medium containing glucose. this increase in the enzyme activity is developed progressively when the concentration of ribose increases from 1% to 6% and it is blocked if cycloheximide is added to the medium. this results suggest an increase in "de novo" synthesis o ...19836418104
energy of binding of aspergillus oryzae beta-glucosidase with the substrate, and the mechanism of its enzymic action.several beta-d-glucopyranosides (p-nitrophenyl, phenyl, and ethyl), 1-thio-beta-d-glucopyranosides, and phenyl 2-deoxy, 3-deoxy, 4-deoxy, and 6-deoxy beta-d-glucopyranosides were synthesized and used to study the mechanism of the enzymatic action of taka-beta-glucosidase [ec aspergillus oryzae]. kinetic constants of the enzyme for these glycosides were determined from s/v-s or 1/v-1/s plots, and the hydrolysis rates of these compounds with the enzyme, acid (3 n hcl) and alkali (3 n naoh ...19836418735
present-day studies on cereals protein nature alpha-amylase inhibitors. 19836190085
a chicken repetitive dna sequence that is highly sensitive to single-strand specific endonucleases.a dna sequence consisting of the 5-mer agagg repeated tandemly 32 times has been detected in a chicken genomic clone and found to be present in about 2000 copies per chicken genome. this sequence was highly susceptible to single-strand specific endonucleases isolated from aspergillus oryzae (s1) and mung bean, but cleavage by a single-strand specific endonuclease isolated from neurospora crassa occurred only at a ph below 5.5. endonucleolytic cutting of the agagg sequence by the single-strand sp ...19836231528
the induction of alpha-amylase by starch in aspergillus oryzae: evidence for controlled mrna expression.the induction of alpha-amylase by starch has been studied in the filamentous fungus aspergillus oryzae. low levels of alpha-amylase activity were found in both intracellular and extracellular samples from glucose-grown cultures. however, alpha-amylase activity increased when starch was the sole carbon source. the intracellular enzyme activity was induced by a factor of approximately 6.5, while the extracellular activity increased 20-fold over that found in the glucose-grown cultures. regardless ...19846208985
antifungal properties of 2-n-alkyn-1-ols.fourteen 2-n-alkynols (c3-c14, c16, and c18) were tested against aspergillus oryzae, aspergillus niger, trichoderma viride, and myrothecium verrucaria in sabouraud dextrose agar at ph 5.6 and 7.0. toxicity to candida albicans, candida tropicalis, trichophyton mentagrophytes, and mucor mucedo was determined in the same medium at ph 5.6 and 7.0 in the absence and presence of 10% beef serum. fungitoxicity was strongly influenced by chain length, slightly by the ph of the medium, and significantly b ...19846098642
multiple forms of protyrosinase from aspergillus oryzae and their mode of activation at ph 3.0.this paper describes the isolation of three molecular forms (i-iii) of protyrosinase and catalytically active tyrosinase ( monophenol ,dihydroxyphenylalanine: oxygen oxidoreductase, e.c. from fresh mycelia of aspergillus oryzae bir 128 by (nh4)2so4 fractionation, successive chromatographies and disc-gel electrophoresis. experimental evidence is presented that purified protyrosinase is contaminated with firmly attached proteinases, and that this contamination may account for both the m ...19846424711
effect of glycolipids contained in liposomes on the incorporation of beta-galactosidase into specific organs.a series of glycolipids were examined to find a system capable of targeting liposomes into specific organs of rats. sulfatide was found to be the best among the components of liposomes examined for delivering the entrapped enzyme, beta-galactosidase from aspergillus oryzae, into the brain and liver; gangliosides, for the spleen; and sphingomyelin, for the lung. to introduce the enzyme into the liver, galactocerebroside was far better than glucocerebroside. these data suggest that the sugar resid ...19846435635
alteration of the substrate specificity of aspergillus oryzae beta-galactosidase by modification with polyethylene glycol.beta-galactosidase (beta-d-galactoside galactohydrolase, ec purified from aspergillus oryzae was modified with 2,4,6-trichloro-s-triazine derivatives of polyethylene glycol (activated bpeg) having molecular weights of 600, 1500, 2000, and 4000. polyethylene glycol derivatives were attached to 6 of the 12 amino groups exposed on the surface of the enzyme. upon modification, the enzymatic activity for a water-soluble substrate, o-nitrophenyl beta-d-galactopyranoside, was reduced with inc ...19846436228
[fatty acid composition of the lipids in fungi of the genus aspergillus developing on mineral media with different carbon sources].this work was aimed at studying the effect of different carbon sources in the composition of mineral media on the growth of fungi belonging to the aspergillus genus and on the fatty acid composition of their lipids. a chemically-defined medium with glucose was shown to be optimal for the growth of 18 aspergillus strains and for the synthesis of lipids by them. the fatty acid composition of lipids was studied when the fungi grew in media with different carbon sources. the lipids were shown to con ...19846442390
[aspergillus oryzae as a cause of keratomycosis in the horse].a case of a spontaneous mycokeratitis of a previously injured cornea in a horse is described. the infection was caused by aspergillus oryzae. after application of chloramphenicol ophthalmic ointment a corneal clouding was found in the centre which was circularly sharply defined and which - after dispensing dexamethason-neomycin eye drops - expanded all over to a purulent keratitis. the demarcated and initially non purulent mycotic lesions largely improved after the application of tincture of iod ...19846528327
[a case of allergic broncho-pulmonary aspergillosis due to asp. oryzae and its subtype]. 19846530860
secondary structure specificity of the nuclease activity of the 1,10-phenanthroline-copper complex.the artificial dnase activity of the 1,10-phenanthroline-cuprous ion complex [(op)2cu+] and h2o2 cleaves the a, b, and z forms of dna at different rates. the b structure, formed by most dnas including poly(da-dt) and poly(da) x poly(dt), is the most susceptible to cleavage. it is completely degraded within 1 min by 40 microm 1,10-phenanthroline/4 microm cu2+/7 mm h2o2/7 mm 3-mercaptopropionic acid. the a structure, formed by rna x dna hybrids such as poly(ra) x poly(dt), is cleaved in both stran ...19846320169
endonuclease s1-sensitive site in chicken pro-alpha 2(i) collagen 5' flanking gene region.a site that is preferentially cleaved by the single-strand-specific endonuclease from aspergillus oryzae was located in vitro 180 base pairs upstream from the 5' end of the chicken pro-alpha 2(i) collagen gene. it is found in supercoiled plasmids with a negative superhelical density of -0.024 or more but not in linear dna molecules. the nuclease s1 sensitivity is retained in plasmids containing genomic fragments extending from position +8 to -285 (where +1 is the first transcribed base) and from ...19846324210
[species and organ heterogeneity of arylsulfatases]. 19846397752
conidium germination rate in wild and domesticated yellow-green aspergilli.conidia of domesticated yellow-green aspergilli from strains of aspergillus oryzae (ahlburg) cohn and aspergillus sojae sakaguchi and yamada ex murakami, used in the preparation of koji inoculum, germinate approximately 3 h sooner than conidia produced by related wild species, aspergillus flavus link ex fr. and aspergillus parasiticus speare. there was no consistent relationship between average conidium size and estimated 50% germination time. germination trials were conducted on czapek agar at ...198416346471
[interrelation of translation regulation and amino acid uptake into cells]. 19853877528
[protease and alpha-amylase synthesis by washed cells of aspergillus oryzae 251-90].regularities of protease and alpha-amylase production by washed cells of aspergillus oryzae 251-90 were being studied. the results obtained enabled us to assume a constitutive character of the both enzymes synthesis by the given producer. sources of carbon, nitrogen and sulphur take part in regulation of the protease production, whereas in the case of the alpha-amylase synthesis only carbon sources that are important. elimination of phosphorus from the medium affects the synthesis of both enzyme ...19853885211
nucleotide sequence of the extracellular glucoamylase gene sta1 in the yeast saccharomyces diastaticus.the complete nucleotide sequence of the extracellular glucoamylase gene sta1 from the yeast saccharomyces diastaticus has been determined. a single open reading frame codes for a 778-amino-acid protein which contains 13 potential n-glycosylation sites. in the 5'- and 3'-flanking regions of the gene, there are striking sequence homologies to the corresponding regions of adh1 for alcohol dehydrogenase and mat alpha 2 for mating type control in the yeast saccharomyces cerevisiae. the putative precu ...19853918017
enzyme replacement with liposomes containing beta-galactosidase from charonia lumpas in murine globoid cell leukodystrophy (twitcher).enzyme replacement with liposomes containing beta-galactosidase obtained from charonia lumpas was carried out in murine globoid cell leukodystrophy (gld). charonia lumpas beta-galactosidase was able to hydrolyze galactocerebroside trapped into liposomes prepared from lecithin, cholesterol and sulfatide (molar ratio; 7:2:1). liposomes containing charonia lumpas beta-galactosidase were successfully incorporated into the mouse tissues. 3h-galactocerebroside labeled liposomes were also incorporated ...19853919736
[mycoses of the orbit and the eye]. 19853922126
effect of additive on alfalfa silage fermentation characteristics and feedlot performance of each of two trials four large plastic bags were filled with approximately 72 mt of first cutting, early bloom alfalfa (35% dry matter). fresh alfalfa in two bags was treated with .5 kg/metric ton of a commercial silage additive (silo-best). two bags were left untreated. when compared with steers fed untreated alfalfa silage, weight gains, intakes, and feed efficiencies were not significantly (p greater than .05) altered for steers fed treated silage. however, laboratory analysis showed silage ...19853928719
purification and characterization of endo-beta-n-acetylglucosaminidase of aspergillus endo-beta-n-acetylglucosaminidase which hydrolyzes the n,n'-diacetylchitobiosyl linkage in asparagine-linked oligosaccharides was purified from the enzyme product of aspergillus oryzae. its substrate specificity was similar to that of endo-beta-n-acetylglucosaminidase h from streptomyces griseus with respect to the relative activities toward the glycopeptides obtained from ovalbumin and bovine igg. the present endoglycosidase exhibited a broad optimum ph range and was relatively stable. metal ...19853934149
evidence that the mechanisms of fungitoxicity of 8-quinolinol and its bischelate with copper(ii) are different.the copper(ii) contents of the growth media, sabouraud dextrose and czapek-dox broths, and of the spore inocula of aspergillus niger (atcc 1004), aspergillus oryzae (atcc 1011), trichoderma viride (atcc 8678), and myrothecium verrucaria (atcc 9095) were determined by atomic absorption spectrophotometry in a graphite furnace. the test systems composed of sabouraud dextrose broth and spore inocula of the four fungi contained only a little over 3% of the copper(ii) required to form a minimal inhibi ...19853935305
the subsite structures of guanine-specific ribonucleases and a guanine-preferential ribonuclease. cleavage of oligoinosinic acids and poly order to estimate the size of the active site of guanylic acid specific rnases (rnase t1 from aspergillus oryzae and rnase st from streptomyces erythreus) and guanine-preferential rnase (rnase ms from a. saitoi), the depolymerization reaction of oligoinosinic acid, (ip)ni greater than p, having various chain lengths was studied. the kinetic parameters for depolymerization of oligoinosinic acids, (pkm, log v and log v/km) by the three rnases increased with increase of the chain length of the s ...19853936847
a novel enzyme producing isoprimeverose from oligoxyloglucans of aspergillus enzyme producing isoprimeverose from xyloglucan fragment oligosaccharides has been purified to the electrophoretically pure state from a commercial enzyme preparation of aspergillus oryzae (sanzyme 1000). the purified enzyme showed approximately 1,280-fold increase of the specific activity over the original preparation. the purified enzyme was shown to be an oligomeric protein consisting of two subunits, each of which had a molecular weight of 115,000. the enzyme showed the highest activity a ...19854019436
antifungal properties of 3-n-alkyn-1-ols and synergism with 2-n-alkyn-1-ols and ketoconazole.twelve 3-n-alkyn-1-ols (c4-c12, c14, c16, and c18) were tested against aspergillus oryzae, aspergillus niger, trichoderma viride, and myrothecium verrucaria in sabouraud dextrose agar at ph 5.6 and 7.0. toxicity to candida albicans, candida tropicalis, trichophyton mentagrophytes, and mucor mucedo was determined in the same medium at ph 5.6 and 7.0 in the absence and presence of 10% beef serum. fungitoxicity was strongly influenced by chain length, slightly by ph of the medium, and significantly ...19854020633
heterokaryosis between aspergillus oryzae cyclopiazonic acid-defective strains: method for estimating the risk of inducing toxin production among cyclopiazonic acid-defective industrial strains.aspergillus oryzae strains are used extensively in the food industry. some of these strains excrete alpha-cyclopiazonic acid (cpa), a mycotoxin which may provoke toxicoses in rats. physicochemical methods may reveal the presence of this toxin, but they are inadequate to screen cpa-nonproducing (cpa-) strains. cpa production is revealed by either bacterial growth inhibition or alkalinization of the culture medium. this first biological property was used to devise a time-saving screening method to ...19854083874
5-iodoribose 1-phosphate, an analog of ribose 1-phosphate. enzymatic synthesis and kinetic studies with enzymes of purine, pyrimidine, and sugar phosphate metabolism.the 5'-deoxy-5'-iodo-substituted analogs of adenosine and inosine are cytotoxic to tumor cells that have high activities of 5'-methylthioadenosine phosphorylase and purine nucleoside phosphorylase, respectively (savarese, t.m., chu, s-h., chu, m.y., and parks, r. e., jr. (1984) biochem. pharmacol. 34, 361-367). 5-iodoribose 1-phosphate (5-irib-1-p), the common intracellular metabolite of these 5'-iodonucleosides, has been synthesized enzymatically from 5'-deoxy-5'-iodoadenosine via adenosine dea ...19862934389
an immunochemical study of neurospora nucleases.nucleases derived from neurospora crassa mycelia with neutral single-strand (ss) endodeoxyribonuclease activity have been examined by immunochemical techniques and by sodium dodecyl sulfate - dna gel electrophoresis. all of the intracellular nucleases, which have different divalent metal ion requirements, different strand specificities with single- and double-strand dna, different modes of action on dna and rna, and other distinguishing characteristics, are immunochemically related to neurospora ...19863013242
production of aflatoxins by strains of the aspergillus flavus group maintained in atcc.a total of 169 strains of the aspergillus reference cultures in the aspergillus flavus group maintained in the american type culture collection (atcc) were studied for their aflatoxin-producing abilities in rice, peanut and semisynthetic medium, utilizing high pressure liquid chromatography. fifty-nine of the strains examined were positive for aflatoxin formation. no strains of the food fungi a. oryzae or a. sojae produced detectable levels of aflatoxins, while 33-85% of the strains of a. flavus ...19863083260
actions of aspergillus oryzae alpha-amylase, potato phosphorylase, and rabbit muscle phosphorylase a and b on phosphorylated (1----4)-alpha-d-glucan.aspergillus oryzae alpha-amylase [(1----4)-alpha-d-glucan glucanohydrolase, ec] produced o-(6-phosphoryl-alpha-d-glucopyranosyl)-(1----4)-o-alpha-d-glucopyran osy l-(1----4)-d-glucopyranose (6(3)-phosphorylmaltotriose) and o-alpha-d-glucopyranosyl-(1----4)-o-(3-phosphoryl-alpha-d-glucopyranosyl )- (1----4)-o-alpha-d-glucopyranosyl-(1----4)-d-glucopyranose (3(3)-phosphorylmaltotetraose) from potato starch upon exhaustive hydrolysis. these products indicate that the enzyme hydrolyses the s ...19863096568
new alpha-amylase and trypsin inhibitors among the cm-proteins of barley (hordeum vulgare).barley cm-proteins are a group of at least five salt-soluble components (cma-e) that can be selectively extracted from endosperm with chloroform/methanol mixtures. n-terminal sequences of proteins cma, cmb and cmc have been determined and found to be homologous to those previously determined for cmd and cme, an observation which confirms that their structural genes are members of a dispersed multi-gene family. the purified cm-proteins were tested against trypsin and against alpha-amylases from s ...19863484638
role of aspergillus amylase in baker's asthma. 19862417074
improved purification of aspergillus oryzae 1,2-alpha-mannosidase by using mannan gel affinity order to facilitate the purification of 1,2-alpha-mannosidase from an enzyme product of aspergillus oryzae, we have devised a rapid and simple procedure. a partially purified enzyme preparation obtained from the a. oryzae enzyme product, by means of ammonium sulfate fractionation followed by cm-sephadex c-50 chromatography, was subjected to affinity chromatography with baker's yeast mannan gel as an adsorbent. 1,2-alpha-mannosidase was retarded and well separated from the major protein peak o ...19863700367
the disulphide folding pathway of ribonuclease t1.the pathway of unfolding and refolding that accompanies, respectively, the breakage and formation of the two disulphide bonds of ribonuclease t1 has been determined kinetically and compared with those of other proteins.19863735429
invasion of paranasal sinuses by aspergillus oryzae.a 24-year-old male patient receiving chemotherapy for acute promyelocytic leukemia developed fever, right periorbital swelling and mild right proptosis. a head scan showed opacification of the right maxillary and ethmoid sinuses with adjacent soft tissue swelling. biopsy of the nasal mucosa demonstrated the typical septate hyphae of aspergillus species which was later shown on culture to be aspergillus oryzae. a. oryzae has only rarely been reported in human disease and there is confusion as to ...19863796709
allergic bronchopulmonary aspergillosis due to aspergillus oryzae.a 19-year-old female student with allergic bronchopulmonary aspergillosis (abpa) due to aspergillus oryzae is reported. this organism was used for fermentation starter to make soybean paste in her father's workshop adjacent to the home where she lived. abpa might be considered an occupational disease in certain situations.19872433099
nucleotide sequence of the alpha-amylase gene (alp1) in the yeast saccharomycopsis fibuligera.the complete nucleotide sequence of the secretable alpha-amylase gene alp1 from the yeast saccharomycopsis fibuligera has been determined. the alp1 dna hybridized to a polyadenylated rna of 2.0 kilobases. a single open reading frame encodes a 494-amino acid protein which is highly homologous with alpha-amylase (taka-amylase) of a fungus aspergillus oryzae.19873497057
differential scanning calorimetric study of the thermal unfolding of taka-amylase a from aspergillus oryzae.the thermally induced unfolding of taka-amylase a, isolated from aspergillus oryzae, was studied by differential scanning calorimetry. the experimental curves of excess apparent specific heat vs. temperature showed a single asymmetric peak. curve resolution indicated that this asymmetry is due to the two-state unfolding of three domains in the molecule, with dissociation of the single tightly bound ca2+ ion occurring during the unfolding of the last domain. further indication of the dissociation ...19873498514
studies on the active site of taka-amylase a: its action on phenyl maltooligosides with a charge at their non-reducing-ends.five modified moltooligosaccharides, phenyl o-6-amino-6-deoxy-alpha-d- glucopyranosyl- (1----4)-o-alpha-d-glucopyranosyl-(1----4)-o-alpha-d-glucopyranosyl-(1-- --4)- alpha-d-glucopyransoide (ag4p), phenyl o-(alpha-d-glucopyranosyluronic acid)-(1----4)-o-alpha-d-glucopyranosyl-(1----4)-o-alpha-d-glucopyran osy l- (1----4)-alpha-d-glucopyranoside (cg4p), phenyl o-6-amino-6-deoxy-alpha-d-glucopyranosyl-(1----4)-o-alpha-d-glucopyra nos yl- (1----4)-o-alpha-d-glucopyranosyl-(1----4)-o-alpha-d-glucopy ...19873501785
effect of yeast culture and aspergillus oryzae fermentation extract on ruminal characteristics and nutrient digestibility.four nonpregnant and nonlactating holstein cows fitted with ruminal fistulas were assigned to each of four diets in a 4 x 4 latin square design. dietary treatments were 1) basal diet containing 50% concentrate; 2) basal diet plus 90 g/d yeast culture; 3) basal diet plus 2.63 g/d aspergillus oryzae fermentation extract; 4) basal diet plus 90 g/d of a. oryzae fermentation extract and yeast culture. cows were fed diets at a rate of 86 g dm/kg bw.75 for 14 d adaptation followed by an 8-d collection ...19873680724
studies of protease and protease inhibitors in familial amyloidotic polyneuropathy.serum levels of 6 protease inhibitors, alpha 1-antitrypsin, cl inactivator, alpha 2-macroglobulin, antithrombin-3, alpha 1-antichymotrypsin and inter-alpha-trypsin inhibitor were measured in patients with familial amyloidotic polyneuropathy (fap) and a control group without neurologic disease. no significant differences were observed between the 2 groups. the proteolytic effect of brinase, an enzyme from aspergillus oryzae, on amyloid tissue sections from patients with fap was also evaluated. am ...19873316509
study on immobilization of aminoacylase from aspergillus oryzae on macroporous copolymers.the aminoacylase from aspergillus oryzae was adsorbed on polymer carriers where the enzyme showed excellent catalyzing activity and operating stability. it was found that the pore structure and functional group were the basic factors which affected the activity of the adsorbed enzyme. the hydrophobility of the carriers did not play an important role in the enzymatic resolution. various factors, such as percentage and nature of cross-linking agent, porogenic agents and functionalizing agents whic ...19873452428
transformation of aspergillus oryzae using the a. niger pyrg gene.a transformation system for aspergillus oryzae based on the orotidine-5'-phosphate decarboxylase gene (pyrg) was developed. transformation frequencies of up to 16 transformants per microgram of dna were obtained with the vector pab4-1, which carries the pyrg gene of a. niger. southern blotting analysis showed that vector dna sequences were integrated into the chromosomal dna, in various copy numbers and presumably at different sites. efficient cotransformation of an unselectable gene was also sh ...19873481025
purification of ribonuclease improved method for purifying ribonuclease t1 from aspergillus oryzae is described. the method uses gradient elution from deae-cellulose and sulfopropyl-sephadex columns followed by gel filtration on sephadex g-50 to give almost 100 mg (50% yield) of ribonuclease t1 from 100 g of starting material in less than 5 days.19873126677
two histidine residues are essential for ribonuclease t1 activity as is the case for ribonuclease a.ribonuclease t1 (rnase t1, ec is a guanosine-specific ribonuclease that cleaves the 3',5'-phosphodiester linkage of single-stranded rna. it is assumed that the reaction is generated by concerted acid-base catalysis between residues glu-58 and his-92 or his-40. from the results of chemical modification and nmr studies, it appeared that the residue glu-58 was indispensable for nucleolytic activity. however, we have recently demonstrated that glu-58 is an important but not an essential re ...19873126807
simple method for screening aflatoxin-producing molds by uv photography.uv absorption by aflatoxins was monitored in gy agar medium by uv photography. in the uv photographs, aflatoxin-producing molds were identified as gray or black colonies, whereas aflatoxin-nonproducing molds appeared as white colonies. by cellophane transplantation experiments and silica gel thin-layer chromatography, the products absorbing uv light substantially were found to be mainly aflatoxins b1 and g1 excreted from the mold mycelium into the agar medium. uv absorption did not occur when th ...19873105453
effective reduction of lactose maldigestion in preschool children by direct addition of beta-galactosidases to milk at mealtime.we examined the efficiency of two beta-galactosidase preparations--one derived from the yeast, kluyveromyces lactis (lactaid), the other derived from the fungus, aspergillus oryzae (takamine)--to assist the in vivo digestion of lactose consumed by healthy guatemalan preschool children. milk prehydrolyzed by in vitro incubation with enzymes was used as the standard of reference, and the degree of incomplete digestion of lactose from 240 ml of milk was determined using the hydrogen breath test. in ...19873106927
facile preparation and some applications of an affinity matrix with a cleavable connector arm containing a disulfide bond.a versatile affinity matrix in which the ligand of interest is linked to the matrix through a connector arm containing a disulfide bond is described. it can be synthesized from any amino-substituted matrix by successive reaction with 2-imino-thiolane, 5,5'-dithiobis(2-nitrobenzoic acid), and a thiol derivative of the ligand of choice. the repertoire of ligands can be significantly increased by the appropriate use of avidin-biotin bridges. after adsorption of the material to be fractionated, elut ...19873110760
does oral enzyme replacement therapy reverse intestinal lactose malabsorption?the effects of oral enzyme replacement therapy on breath hydrogen excretion and symptoms after milk ingestion were studied in lactase-deficient patients. sixteen symptomatic patients underwent interval hydrogen breath tests using whole milk as substrate. each study was repeated with the addition of 250 mg of beta-d-galactosidase derived from aspergillus oryzae (lactrase) given orally with the milk. subsequently seven of those 11 patients who did not normalize their hydrogen excretion with 250 mg ...19873111243
protein dynamics. a time-resolved fluorescence, energetic and molecular dynamics study of ribonuclease t1.studies using time-resolved fluorescence depolarization were performed on the internal motion of trp 59 of ribonuclease t1 (ec in the free enzyme, 2'-gmp-enzyme complex and 3'-gmp-enzyme complex. the trp 59 motion was also studied in the free enzyme using molecular dynamics simulations. energetic analysis of activation barriers to the trp 59 motion was performed using both the transition state theory and kramers' theory. the activation parameters showed a dependence on solvent viscosit ...19873111558
frequency domain measurements of the fluorescence lifetime of ribonuclease t1.using multifrequency phase/modulation fluorometry, we have studied the fluorescence decay of the single tryptophan residue of ribonuclease t1 (rnase t1). at neutral ph (7.4) we find that the decay is a double exponential (tau 1 = 3.74 ns, tau 2 = 1.06 ns, f1 = 0.945), in agreement with results from pulsed fluorometry. at ph 5.5 the decay is well described by a single decay time (tau = 3.8 ns). alternatively, we have fitted the frequency domain data by a distribution of lifetimes. temperature dep ...19873115328
structures of oligosaccharides on beta-galactosidase from aspergillus oryzae.the carbohydrate portions of beta-galactosidase from aspergillus oryzae were found to be composed of two types of sugar chains. they were released equally well with endo-beta-n-acetylglucosaminidase h, but were distinct in their chain length. the long sugar chains (fraction i), corresponding to 4% of the total carbohydrate chains, were composed of galactomannan-type oligosaccharides, which consisted of mannose, galactose, glucose, and glucosamine in the molar ratios of 30.0, 16.4, 1.4, and 2.1 p ...19873117779
[enzymatic isolation of the pyramidal neurons of the rat hippocampus]. 19873034680
[development of new plate tests for the detection of microbial hydrolysis of esters and oxidation of 2-hydroxycarboxylic acids].the application of the thin-agar-layer coated filter culture technique (z. naturforsch. 42c, 1082 (1987)) has been extended to the detection of ester hydrolysis and 2-hydroxyacid oxidation by microbial colonies. the former was performed by spraying with bromocresol purple and the latter with a salicylhydrazide reagent. under the optimized conditions, the hydrolysis activity of more than 20 mumol/h.g-wet cells and the oxidation activity of more than 40 mumol/h.g-wet cells were usually detected di ...19872966500
immobilization of single-strand specific nuclease (s1 nuclease) from aspergillus oryzae.s1 nuclease from aspergillus oryzae (ec was coupled to gelatin-alginate composite matrix using the residual free aldehyde groups on the surface of glutaraldehyde crosslinked matrix. the immobilized enzyme retained approximately 10% activity of the soluble enzyme. when partially purified enzyme was bound to the matrix, the immobilized preparation did not show any detectable enzyme activity. however, the activity could be restored when the coupling was carried out in the presence of a co ...19872820305
action of gamma endonuclease on clustered lesions in irradiated dna.irradiation of dna in situ i.e. in phage particles or in the cell leads to alterations of single dna nucleotides as well as to clustered lesions such as double strand breaks or unpaired dna regions the latter being sensitive to digestion by s1 nuclease. a contribution will be made to the configuration of such s1-nuclease-sensitive sites (s1 sites). dna from irradiated lambda phage containing s1 sites was treated with gamma endonuclease from m. luteus which is known to split the nucleotide strand ...19882839862
1h-nmr investigation of the interaction between rnase t1 and a novel substrate analog, 2'-deoxy-2'-fluoroguanylyl-(3'-5')uridine.the interaction between rnase t1 and a non-hydrolysable substrate analog, 2'-deoxy-2'-fluoroguanylyl-(3'-5')uridine (gfpu), was investigated using 1h-nmr spectroscopy. in the complex, the gfp portion takes the syn form around the glycosidic bond and the 3'-endo form for the ribose moiety, similar to those found in 3'-gmp and 2'-deoxy-2'-fluoroguanosine 3'-monophosphate (gfp). however, in contrast to the cases of these two inhibitors, the complex formation with gfpu at ph 6.0 was found to shift t ...19882841155
jokichi takamine (1854-1922). 19883057095
purification and properties of an extracellular glucoamylase from a diastatic strain of saccharomyces cerevisiae.the extracellular glucoamylase from certain strains of saccharomyces cerevisiae can be purified from culture medium by a simple chromatographic procedure. the native enzyme is heavily glycosylated and has an mr of about 250,000, but gel filtration indicates the existence of oligomers of larger size. dissociation yields a form of mr about 70,000. the glucoamylase is rich in serine and threonine and in aspartic acid plus asparagine, and has a pi of 4.62 and a ph optimum of 4.5-6.5. the thermostabi ...19883124820
expression of the chemically synthesized gene for ribonuclease t1 in escherichia coli using a secretion cloning vector.the gene for ribonuclease t1 from aspergillus oryzae has been chemically synthesized using the segmental support technique. an escherichia coli clone producing the ribonuclease at high levels was constructed by linking the gene downstream to the region coding for the signal peptide of the ompa protein (a major outer membrane protein of e. coli), using the secretion cloning vector pin-iii-ompa2. this strategy was employed in order to circumvent a possible toxic effect of the gene product on the h ...19883131142
molecular dynamics simulations of ribonuclease t1: analysis of the effect of solvent on the structure, fluctuations, and active site of the free enzyme.molecular dynamics simulations were performed on ribonuclease t1 (rnase t1; ec to determine a structure for the free enzyme. simulations starting with the x-ray coordinates for the 2'gmp-rnase t1 complex were done in vacuo and with an 18-a water ball around the active site using stochastic boundary conditions to understand the influence of water on both the structure and fluctuations of the enzyme. removal of 2'gmp caused structural changes in the loop regions, including those directly ...19883139027
two-dimensional 1h-nmr investigation of ribonuclease t1. resonance assignments, secondary and low-resolution tertiary structures of ribonuclease t1.ribonuclease t1 was studied by two-dimensional 1h-nmr spectroscopy. resonance assignments were obtained for the backbone protons of 95 amino acid residues and most of its side-chain protons using sequence-specific assignment procedures. the secondary structure elements of ribonuclease t1 were identified by an investigation of medium- and long-range nuclear overhauser effects between the backbone and c beta protons. a low-resolution three-dimensional structure of ribonuclease t1 was deduced from ...19883143569
mycological identification of pulmonary aspergilloma caused by aspergillus oryzae with proliferating heads. 19883148401
amino-acid sequence of ribonuclease t2 from aspergillus oryzae.the amino acid sequence of ribonuclease t2 (rnase t2) from aspergillus oryzae has been determined. this has been achieved by analyzing peptides obtained by digestions with achromobacter lyticus protease i, staphylococcus aureus v8 protease, and alpha-chymotrypsin of two large cyanogen bromide peptides derived from the reduced and s-carboxymethylated or s-aminoethylated protein. digestion with a. lyticus protease i was successfully used to degrade the n-terminal half of the s-aminoethylated prote ...19883169020
activation of antioxidant activity in natural medicinal products by heating, brewing and lipophilization. a new drug delivery system.on the basis of the authors' previous hypothesis (1) that treatment of natural plant products with heating and gastric juice results in liberation of low molecular weight compounds with antioxidant activity that exist in the native products as repeating subunits of high molecular weight polymers, they have developed specially treated natural health or medicinal products in which several identifiable low molecular weight compounds with antioxidant activity are liberated and activated. the princip ...19883219997
[the role of alpha-amylase in baker's asthma].we report a case of asthma in a baker provoked by an additive which is generally used in bread making, namely alpha-amylase, an extract of aspergillus oryzae. skin tests to alpha-amylase were positive in a concentration of 0.1 micrograms/ml. specific ige was detected to this enzyme. a provocation test to alpha-amylase was positive with a fall of the fev1 of 42%. provocation tests to flour gave a negative immediate and delayed reaction. progress was made towards the disappearance of symptoms afte ...19883263674
[comparison of chemical and thermal stability of soluble and immobilized alpha-amylases from b. licheniformis and a. oryzae].alpha-amylases from b. licheniformis and a. oryzae, which differ in their thermal stability, were studied with respect to their resistance to chemical denaturants as guanidine hydrochloride, urea, aliphatic alcohols, and formaldehyde. moreover, the thermal and chemical stabilities of both enzymes were compared after their covalent binding to gamma-aminopropyl silica. the thermolabile alpha-amylase (a. oryzae) as well as the thermostable alpha-amylase (b. licheniformis) are remarkably stabilized ...19883266840
alpha-amylase structure and activity.two computerized methods of predicting protein secondary structure from amino acid sequences are evaluated by using them on the alpha-amylase of aspergillus oryzae, for which the three-dimensional structure has been determined. the methods are then used, with amino acid alignments, to predict the structures of other alpha-amylases. it is found that all alpha-amylases of known amino acid sequence have the same basic structure, a barrel of eight parallel stretches of extended chain surrounded by e ...19883267138
nutritive value of whole soybeans fermented with aspergillus oryzae or rhizopus oligosporus as evaluated by neonatal pigs.two experiments were conducted to evaluate the nutritive effects of heated whole soybeans fermented with the mold aspergillus oryzae (aos) or rhizopus oligosporus (ros) in diets fed to neonatal pigs from 1 to 22 d of age. in experiment 1, either soybean meal or heated whole soybeans fermented with ros replaced 75% of the dried skim milk protein. the average daily gain (adg) and gain-feed (g-f) ratio of pigs fed the diet containing 100% milk protein were greater than those of pigs fed the diets c ...19883357058
[changes in the composition and unsaturated state of the neutral lipid fraction during the development of the fungus aspergillus oryzae]. 19883412198
properties of the complex between alpha 2-macroglobulin and brinase, a proteinase from aspergillus oryzae with thrombolytic effect.the proteinase, brinase (mr approximately 35000), from aspergillus oryzae, which has been used in therapeutic attempts as a thrombolytic agent in arterial thrombosis, binds to purified human alpha 2-macroglobulin (alpha 2m) with a stoichiometry of 1.7-1.9 mol of enzyme/mol inhibitor. this binding leads to quantitative cleavage of the bait region of the inhibitor and to release of 3.6 thiol groups per molecule of alpha 2m, reflecting cleavage of the thioester bonds. the reaction with brinase is a ...19882450410
conformational stability and activity of ribonuclease t1 with zero, one, and two intact disulfide bonds.ribonuclease t1 has two disulfide bonds linking cysteine residues 2-10 and 6-103. we have prepared a derivative of ribonuclease t1 in which one disulfide bond is broken and the cysteine residues carboxymethylated, (2-10)-rcm-t1, and three derivatives in which both disulfides are broken and the cysteine residues reduced, r-t1, carboxamidomethylated, rcam-t1, or carboxymethylated, rcm-t1. the rna hydrolyzing activity of these proteins has been measured, and urea and thermal denaturation studies ha ...19882457027
biosynthesis, in calf pancreas microsomes, of three lipid-linked oligosaccharide diphosphates from a synthetic dolichyl diphosphate tetrasaccharide.incubation of calf pancreas microsomes with synthetic alpha-d-manp-(1----6)-beta-d-manp-(1----4)-beta-d-glcpnac-(1 ----4)-alpha-d- glcpnac-pp-dol and gdp-d-[14c]-mannose gave three major lipid-linked oligosaccharide diphosphates. after release of the phospholipid residue by mild acid hydrolysis, the corresponding [14c]oligosaccharides were analyzed by gel-filtration, liquid chromatography, degradation by endo-n-acetyl-beta-d-glucosaminidases d and h, by jack bean alpha-d-mannosidase and aspergil ...19882463085
genetic transformation of an argb mutant of aspergillus argb mutant of aspergillus oryzae nrrl 492 has been genetically transformed with the aspergillus nidulans argb gene. protoplasts were generated with a combination of novozyme 234 and beta-glucuronidase and regenerated on sucrose-stabilized minimal medium without arginine as described for a. nidulans. a frequency of 5 to 10 transformants per mug of dna was obtained; however, most transformants appeared abortive. the a. nidulans argb gene and vector sequences appeared to be integrated into the ...198816347669
uptake of glycosylated ?-galactosidase into human brain synaptosomes: occurrence of two different systems for specific glycosides.uptake of glycoproteins into brain nerve terminals was studied, using glycosylated ?-galactosidase and human brain synaptosomes. a series of p-aminophenyl ?- or ?-glycosides were covalently bound to ?-galactosidase purified from aspergillus oryzae and uptake of the enzyme into human brain synaptosomes was measured. ?-galactosidase coupled with ?-d-mannoside (?-d-man ?-gal) was the best to be taken up among glycoconjugates tested. the uptake was dependent on the amount of ?-d-man ?-gal, the incub ...198820501274
hydrolysis of lactose in skim milk by immobilized beta-galactosidase in a spiral flow reactor.beta-galactosidase from aspergillus oryzae immobilized in a spiral flow reactor was used to effect the hydrolysis of the lactose component of skim milk. residence time distribution measurements were used to assess the amount of longitudinal dispersion occurring as a consequence of the spiral flow pattern and the semiporous nature of the polymeric material used to construct the spiral. it was possible to model the flow conditions as tubular flow with a peclet number that was a linear function of ...198918587857
hydrophobic cluster analysis of the primary sequences of alpha-amylases.the amino acid sequences of 18 alpha-amylases have been compared by hydrophobic cluster analysis. the method was first calibrated with two alpha-amylases (aspergillus oryzae and pig pancreas) whose three-dimensional structures are known. it was then applied to the other alpha-amylases resulting in straightforward sequence alignments which could be used for structure prediction. it was found that all alpha-amylases which were investigated display the same basic super-secondary structure with a (b ...19892489084
enzyme replacement for lactose malabsorption using a beta-d-galactosidase.we evaluated 10 healthy symptomatic lactose malabsorbers for effect of an oral beta-d-galactosidase derived from aspergillus oryzae (lactrase, kremers urban company, milwaukee, wi, u.s.a.) on symptom and breath hydrogen response to challenge with 50 g lactose. basally and at 30-min intervals for 8 h after lactose challenge, end-alveolar breath samples were collected and analyzed for hydrogen using gas chromatography. symptoms were scored at 30 min and hourly for 8 h, rating bloating, cramps, nau ...19892502573
binding of vanadate (v) to ribonuclease-t1 and inosine, investigated by 51v nmr spectroscopy.ribonuclease t1 (rnase-t1) from aspergillus oryzae cleaves ribonucleic acid specifically at guanosine to yield oligonucleotides with terminal guanosine-3'-phosphate. it forms a complex with vanadate (association constant k approximately 145 +/- 30 m-1; delta (51v) = -514 ppm) with spectral features similar to the less stable complexes obtained with di- and tripeptides (gly-his, pros-his-ala, gly-his-lys, val-glu) containing amino acids that are constituents at the active site of the enzyme. guan ...19892513377
synergistic antifungal action of 8-quinolinol and its bischelate with copper(ii) and with mixed ligand chelates composed of copper(ii), 8-quinolinol, and aromatic hydroxy acids.antifungal studies were made of mixtures of minimal inhibitory concentrations (mics) of 8-quinolinol and its bischelates with copper(ii), zinc(ii), and manganese(ii) and with mixed ligand chelates composed of 8-quinolinol, copper(ii) and a second ligand including salicylic acid, 3-hydroxy-2-naphthoic acid, 3,5-diiodosalicylic acid, and 4-bromo-3-hydroxy-2-naphthoic acid. mixtures of the mics of the bischelates of 8-quinolinol with copper(ii) and zinc(ii) and copper(ii) and manganese(ii), as well ...19892516124
three-dimensional structure of ribonuclease t1 complexed with guanylyl-2',5'-guanosine at 1.8 a resolution.the enzyme ribonuclease t1 (rnase t1) isolated from aspergillus oryzae was cocrystallized with the specific inhibitor guanylyl-2',5'-guanosine (2',5'-gpg) and the structure refined by the stereochemically restrained least-squares refinement method to a crystallographic r-factor of 14.9% for x-ray data above 3 sigma in the resolution range 6 to 1.8 a. the refined model consists of 781 protein atoms, 43 inhibitor atoms in a major site and 29 inhibitor atoms in a minor site, 107 water oxygen atoms, ...19892541256
rhizomucor miehei triglyceride lipase is processed and secreted from transformed aspergillus oryzae.the cdna encoding the precursor of the rhizomucor miehei triglyceride lipase was inserted in an aspergillus oryzae expression vector. in this vector the expression of the lipase cdna is under control of the aspergillus oryzae alpha-amylase gene promoter and the aspergillus niger glucoamylase gene terminator. the recombinant plasmid was introduced into aspergillus oryzae, and transformed colonies were selected and screened for lipase expression. lipase-positive transformants were grown in a small ...19892586234
[the stimulating effect of n-nitroso-ethylurea on the biosynthetic ability of aspergillus oryzae].the mutagenic action of n-nitroso-n-ethylurea (neu) taken at shock and prolonged doses together on aspergillus oryzae yielded 98% of populations with an elevated synthesis of proteolytic enzymes. the combined action of shock and prolonged neu doses had an advantage over a shock pulse dose because the frequency of mutations rose 2-16 times and the populations accumulated proteolytic enzymes within the range of 9 to 24 activity units. as was shown using a certain number of populations selected at ...19892586348
preparation and properties of rnase t2 immobilized on concanavalin a-sepharose.partially purified rnase t2 (ec from aspergillus oryzae was bound through its carbohydrate moiety to concanavalin a-sepharose. the retention of activity was high, ranging from 70% at low enzyme load to approximately 9% at high enzyme load. though there was no change in the ph and temperature optima, the ph stability and the km decreased after immobilization. compared to the soluble enzyme, the immobilized rnase t2 showed enhanced temperature stability and more resistance to metal ions. ...19892596845
purification and characterization of an insect alpha-amylase inhibitor/endochitinase from seeds of job's tears (coix lachryma-jobi).a protein inhibitor of locust gut alpha-amylase was purified from seeds of job's tears (coix lachryma-jobi) using ammonium sulphate precipitation, affinity chromatography on red sepharose and reversed-phase hplc. it consisted of two major isomers, each a dimer of two closely similar or identical subunits of mr about 26,400, and associated by inter-chain disulphide bonds. these isomers also had closely similar amino acid compositions. the major isomer showed no inhibitory activity against amylase ...19892605263
isolation of a cdna encoding aspergillus oryzae taka-amylase a: evidence for multiple related genes.complementary and genomic dnas encoding aspergillus oryzae taka-amylase a (taa) were cloned and sequenced. the coding sequence of the cdna comprised the signal peptide [21 amino acids (aa)] and mature taa (478 aa). the deduced aa sequence agrees well with the published aa sequence, except for one insertion, one deletion and ten aa substitutions. these differences might be due to the difference in the strains used. sequence comparison of the cdna and genomic dna indicates the presence of eight in ...19892612911
characterization of three enzymatic forms of glucose-6-phosphate dehydrogenase from aspergillus oryzae.three forms (i, ii and iii) of glucose-6-phosphate dehydrogenase were isolated from mycelium of aspergillus oryzae grown on ribose as the carbon source, by ion-exchange chromatography. the km values determined for the three forms with respect to glucose-6-phosphate were nearly identical; however the km for nadp+ were different and the vmax for the isoenzymatic form ii was higher than those for i and iii. inhibition by nadph was competitive with respect to nadp+, isoenzyme ii showing the highest ...19892616874
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 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
a gene transfer system based on the homologous pyrg gene and efficient expression of bacterial genes in aspergillus oryzae.a homologous transformation system for aspergillus oryzae is described. the system is based on an a. oryzae strain deficient in orotidine-5'-phosphate decarboxylase (pyrg) and the vector pao4-2, which contains a functional a. oryzae pyrg gene as selection marker. transformation of the a. oryzae pyrg mutant with circular pao4-2 resulted in the appearance of pyr+ transformants at a frequency of up to 20 per micrograms of dna, whereas with linear pao4-2 up to 200 transformants per micrograms dna we ...19892688930
a full length cdna clone for the alkaline protease from aspergillus oryzae: structural analysis and expression in saccharomyces cerevisiae.we have cloned and determined the nucleotide sequence of a cdna fragment for the entire coding region of the alkaline protease (alp) from a filamentous ascomycete aspergillus oryzae. according to the deduced amino acid sequence, alp has a putative prepro region of 121 amino acids preceding the mature region, which consists of 282 amino acids. a consensus sequence of a signal peptide consisting of 21 amino acids is found at the n-terminus of the prepro region. the primary structure of the mature ...19892693947
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
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