| studies on the active site of ribonucleases from aspergillus saitoi (author's transl). | | 1976 | 9458 |
| the structure and function of acid proteases. v. comparative studies on the specific inhibition of acid proteases by diazoacetyl-dl-norleucine methyl ester, 1,2-epoxy-3-(p-nitrophenoxy) propane and pepstatin. | comparative studies have been made on the effects of diazoacetyl-dl-norleucine methyl ester (dan), 1,2-epoxy-3-(p-nitrophenoxy)propane (epnp) and pepstatin on acid proteases, including those from acrocylindrium sp., aspergillus niger, aspergillus saitoi, mucor pusillus, paecilomyces varioti, rhizopus chinensis, and trametes sanguinea, and also porcine pepsin ec 3.4.23.1 and calf rennin ec 3.4.23.4 for comparative purposes. these enzymes were rapidly inactivated at similar rates and in 1:1 stioch ... | 1976 | 10290 |
| circular dichroism studies on the n-bromosuccinimide oxidation of ribonuclease from aspergillus saitoi. | | 1977 | 18443 |
| c-terminal peptidyl-l-proline hydrolase activity of aspergillus acid carboxypeptidase. | aspergillus saitoi acid carboxypeptidase hydrolyzed c-terminal peptidyl-l-proline bonds and released the c-terminal proline from z-gly-pro-leu-gly-pro and z-gly-pro at ph 3.3. proline liberated by the enzymic reaction was measured by a sensitive colorimetric ninhydrin method in glacial acetic acid at 513 nm. a km value of 1.0 mm and a kcat value of 0.09 s-1 for z-gly-pro-leu-gly-pro hydrolysis, and a km value of 5.0 mm and a kcat value of 0.0045 s-1 for z-gly-pro hydrolysis were calculated from ... | 1977 | 19444 |
| purification of an acid proteinase from aspergillus saitoi and determination of peptide bond specificity. | the specificity and mode of action of an acid proteinase (ec 3.4.23.6) from aspergillus saitoi were investigated with oxidized b-chain of insulin, angiotensin ii and bradykinin. further purification of acid proteinase was performed with n,o-dibenzyloxycarbonyl-tyrosine hexamethylene-diamino-sepharose 4b affinity chromatography and isoelectric focusing. the purified enzyme was free of any other proteolytic activity demonstrated in asp. saitoi. acid proteinase from asp. saitoi hydrolyzed primarily ... | 1977 | 21699 |
| photooxidation and carbethoxylation of a minor ribonuclease from aspergillus saitoi. | in order to investigate the nature of amino acid residues involved in the active in the active site of a ribonuclease from aspergillus saitoi, the ph dependence of the rates of inactivation of rnase ms by photooxidation and modification with diethylpyrocarbonate were studied. (1) rnase ms was inactivated by illumination in the presence of methylene blue at various ph's. the ph dependence of the rate of photooxidative inactivation of rnase ms indicated that at least one functional group having pk ... | 1977 | 23378 |
| ph-profiles of the kinetic parameters of a minor ribonuclease from aspergillus saitoi. | | 1978 | 25272 |
| immobilization of aspergillus beta-glucosidase on chitosan. | beta-glucosidase of aspergillus phoenicis qm 329 was immobilized on chitosan, using the bifunctional agent glutaraldehyde. the most active preparation based on the amount of support contained a 1:2.5 enzyme-to-chitosan ratio (wt/wt). however, the specific activity of the bound enzyme decreased from 10 to 1% with increasing enzyme-to-chitosan ratio. compared with free beta-glucosidase, the immobilized enzyme exhibited: (i) a similar ph optimum but more activity at lower ph values; (ii) improved t ... | 1978 | 25624 |
| purification and properties of a new ribonuclease from aspergillus saitoi. | from a commercial digestive produced from aspergillus saitoi, a ribonuclease [ec 3.1.4.23] having a molecular weight of 12,500 has been isolated in addition to the rnase reported previously, which had a molecular weight of 38,000. the enzyme was found to be homogeneous by chromatography on deae-cellulose, disc electrophoresis on polyacrylamide gel, and ultracentrifugation. the nh2-terminal amino acid was identified as glutamic acid. the amino acid composition indicated the presence of about 13 t ... | 1975 | 239932 |
| modification of an arginine residue of a base-nonspecific ribonuclease from aspergillus saitoi. | 1. a base-nonspecific ribonuclease from aspergillus saitoi [rnase ms, ec 3.1.4.23; molecular weight, 12,500] was modified with phenylglyoxal (pg) and 1,2-cyclohexanedione (chd) in order to determine whether a single arginine residue was involved in the active site of the enzyme. 2. rnase ms was inactivated by both pg and chd with concomitant loss of one arginine residue. a competitive inhibitor of rnase ms, 2',(3')-amp, protected the enzyme from inactivation by pg. these findings strongly sugges ... | 1979 | 447619 |
| carboxymethylation of a minor ribonuclease from aspergillus saitoi. | (1) rnase ms was inactivated by iodoacetate. the inactivation was most rapid at ph 6.0, and was inhibited in the presence of a denaturant such as 8 m urea or 6 m guanidine-hcl. (2) competitive inhibitors protected rnase ms from inactivation by iodoacetate; the effect was in the order 2',(3')-gtp greater than 2',(3')-amp, 2',(3')-ump greater than or equal to 2',(3')-cmp. the order is not consistent with that of the binding constants of the 4 nucleotides towards rnase ms (a is greater than c great ... | 1979 | 479129 |
| the effect of elimination of amino acids from the carboxyl-terminal of the minor ribonuclease from aspergillus saitoi by carboxypeptidase a on the enzymatic activity. | | 1978 | 639178 |
| the effect of modification of amino-groups of the minor ribonuclease from aspergillus saitoi on the enzymatic activity. | | 1977 | 924992 |
| further studies on the specificity of the minor ribonuclease from aspergillus saitoi. | in order to investigate the base specificity of the minor rnase [ec 3.1.4.23] from aspergillus saitoi, the kinetic constant of the enzyme was measured with 16 dinucleoside phosphates (xpy's) as substrates at ph 5.5 and 25 degrees. the maximum rates of transesterification of gpy's were in the range of 10,000 to 2,800 and were markedly larger than those of other xpy's, including xpg's. the average km values of upy, cpy, apy, and gpy increased in the order a, c, u, and g. this order coincides with ... | 1976 | 965365 |
| purification and characterization of a strictly specific beta-d-fucosidase from aspergillus phoenicis. | although beta-d-fucosidase (beta-d-fucohydrolase, ec 3.2.1.38) has been isolated from various sources, all those enzymes were associated with a high activity of beta-d-galactosidase and/or beta-d-glucosidase. we have purified a specific beta-d-fucosidase in electrophoretically homogeneous form from crude extracts of aspergillus phoenicis by polyethyleneglycol 8000-phosphate buffer aqueous two-phase separation, and successive chromatography on deae-sephadex a-50, hydroxyapatite, and sephadex g-10 ... | 1992 | 1524431 |
| [studies on the beta-d-fucosidase from aspergillus phoenicis]. | although beta-d-fucosidase (beta-d-fucoside fucohydrolase, ec 3.2.1.38) has been isolated from various sources, the identity of this enzyme is still not settled. we have purified a specific beta-d-fucosidase in electrophoretically homogeneous form crude extracts of aspergillus phoenicis by polyethyleneglycol 6000-phosphate buffer aqueous two-phase separation, and successive chromatography on deae-sephadex a-50, hydroxyapatite and sephadex g-100 columns. the molecular weight of the enzyme was est ... | 1992 | 1598757 |
| primary structure of a base non-specific and adenylic acid preferential ribonuclease from aspergillus saitoi. | the complete primary structure of a base non-specific and adenylic acid preferential rnase (rnase m) from aspergillus saitoi was determined. the sequence was determined by analysis of the peptides generated by digestion of heat-denatured rnase m with lysylendopeptidase, and the peptides generated from rcm rnase m by digestion with staphylococcal v8 protease or chemical cleavage with brcn. it consisted of 238 amino acid residues and carbohydrate moiety attached to the 74th asparagine residue. the ... | 1990 | 2229029 |
| [purification and properties of beta-glucosidase from aspergillus phoenicis]. | a beta-glucosidase has been purified to electrophoretically homogeneity from the wheat bran culture of aspergillus phoenicis by peg 6000-phosphate biphasic separation, column chromatography on sephadex g-100, deae-sephadex a-50 and se-sephadex c-50. the enzyme showed optimal activity at ph 5.0 and 60 degrees c. it was stable in the ph range of 4.0-7.5 and up to 55 degrees c. the enzyme activity was strongly inhibited by ag+ and hg2+. the molecular weight of the enzyme was 118000 as determined by ... | 1989 | 2506700 |
| crystallization of a complex between ribonuclease ms and 3'-guanylic acid. | the crystals of a complex between ribonuclease ms, the extracellular ribonuclease from aspergillus saitoi, and 3'-guanylic acid were obtained from 2-methyl-2,4-pentanediol solution by vapor diffusion technique in the hanging drop mode. the crystals belong to orthorhombic space group p2(1)2(1)2(1) with dimensions a = 47.0 a, b = 62.8 a, c = 37.9 a. the crystals diffract strongly up to at least 2.0 a resolution. | 1989 | 2547976 |
| recognition pattern of different bases in the active site of ribonuclease ms--a model building study. | the structure of base non-specific ribonuclease ms from aspergillus saitoi was predicted by sequence similarity to guanine-specific rnase t1 of known structure. in this paper the interaction pattern of binding site of rnase ms with different nucleic acids bases is analysed using model building and energy minimisation techniques. it is shown that unspecificity of this protein can be explained only when taking into account flexibility of the base recognition loop. | 1989 | 2604906 |
| the tertiary structure of aspergillus saitoi minor ribonuclease (ms) predicted from the structure of rnase t1. | ribonuclease ms from aspergillus saitoi is a small acidic protein (11,714 da) containing 106 amino acids of known sequence. unlike other enzymes belonging to the rnase t1 family this ribonuclease is base-unspecific. using interactive computer graphics and energy minimisation we predicted the structure of rnase ms on the basis of sequence homology to rnase t1 of known structure. in this report the predicted structure of this protein is presented and characterised. | 1988 | 3142794 |
| [purification and properties of xylanases from aspergillus phoenicis]. | | 1987 | 3448814 |
| [substrate specificities of lichenase and xylanases from aspergillus phoenicis]. | | 1987 | 3448815 |
| alpha-mannosidases i and ii from aspergillus saitoi. | | 1987 | 3600352 |
| purification and properties of a phospholipase c that has high activity toward sphingomyelin from aspergillus saitoi. | an enzyme hydrolyzing sphingomyelin was purified from extracts of solid cultures of aspergillus saitoi 7041 by fractionation with isopropanol followed by successive column chromatographies on deae-sepharose cl-6b, butyl-toyopearl 650 m, and phenyl-sepharose cl-4b. the preparation of purified enzyme was homogeneous and had an activity increased 81-fold over that of the isopropanol fraction. the yield was about 65%. the molecular weight was estimated to be 54,000 by sodium dodecyl sulfate-gel elec ... | 1987 | 3675875 |
| site of alkylation of the major ribonuclease from aspergillus saitoi with iodoacetate. | a base non-specific and adenylic acid preferential ribonuclease from aspergillus saitoi (rnase m) was modified by [14c]iodoacetic acid. rnase m was inactivated with concomitant incorporation of about 1 mol equivalent of carboxymethyl group. carboxymethylated rnase m (cm rnase m) thus obtained was reduced and carboxymethylated (rcm cm rnase m). from tryptic and chymotryptic digests of rcm cm rnase m, two carboxymethylated histidine-containing peptides labeled with radioactivity were isolated. the ... | 1986 | 3711038 |
| purification and characterization of a novel alpha-mannosidase from aspergillus saitoi. | an alpha-mannosidase differing from 1,2-alpha-mannosidase was found to occur in aspergillus saitoi. by a series of column chromatographies the enzyme was purified up to 1,000-fold, and its properties were studied in detail. the enzyme preparation, which was practically free from other exoglycosidases, showed a ph optimum of 5.0. in contrast to 1,2-alpha-mannosidase, the enzyme was strongly activated by ca2+ ions. p-nitrophenyl alpha-mannopyranoside was not hydrolyzed by the enzyme. accordingly, ... | 1986 | 3745139 |
| modification of a minor glucoamylase from aspergillus saitoi with 1-cyclohexyl-3-(2-morpholinyl-(4)-ethyl)carbodiimide metho p-toluenesulfonate. | in order to elucidate the structure-function relationship of glucoamylases [ec 3.2.1.3, alpha-d-(1-4)-glucan glucohydrolase] from aspergillus saitoi, the reaction of a minor component, gluc m2 with 1-cyclohexyl-3-(2-morpholinyl-(4)-ethyl)carbodiimide metho p-toluenesulfonate (cmc) was studied at ph 4.5. inactivation of gluc m2 with [14c]cmc proceeded with the incorporation of about 5 cmc moieties. from the results of analyses of amino acid and sulfhydryl contents of cmc-modified gluc m2 and the ... | 1985 | 3924906 |
| inhibition of glucoamylases from a rhizopus sp. and aspergillus saitoi by aminoalcohol derivatives. | the mechanism of inhibition of the two glucoamylases from a rhizopus sp. and aspergillus saitoi by aminoalcohol derivatives was investigated. hydrolysis of maltose by the glucoamylases was inhibited competitively by aminoalcohols at ph 5.0, and tris(hydroxymethyl)aminomethane, 2-amino-2-ethyl-1,3-propanediol and 2-aminocyclohexanol were relatively good inhibitors of the glucoamylases among the aminoalcohol derivatives tested. one hydroxyl group and an amino group in these inhibitors were indispe ... | 1985 | 3934147 |
| specificity and mode of action of acid carboxypeptidase from aspergillus saitoi. | | 1973 | 4351261 |
| some properties of ribonucleases from aspergillus saitoi and seminal vesicles immobilized on sepharose 4b activated by cyanogen bromide. | | 1974 | 4468103 |
| mode of action on proteins of acid carboxypeptidase from aspergillus saitoi. | | 1974 | 4474166 |
| submerged production, purification, and crystallization of acid carboxypeptidase from penicillium janthinellum ifo-8070. | penicillium janthinellum ifo-8070 produced an acid carboxypeptidase of molecular weight 51,000 in a liquid medium at 25 c. maximum enzyme concentration was obtained within 3 to 6 days in a medium containing 2% wheat bran, 1% defatted soybean, and 1% kh(2)po(4); the initial ph was 2 to 4. when submerged aerobic conditions were used, a 51,000-molecular-weight acid carboxypeptidase was produced and no detectable amounts of 160,000-molecular-weight acid carboxypeptidase were produced. acid carboxype ... | 1974 | 4474829 |
| inactivation of acid proteases from rhizopus chinensis, aspergillus saitoi and mucor pusillus, and calf rennin by diazoactylnorleucine methyl ester. | | 1972 | 4552474 |
| studies on the state of tryptophan residues in ribonuclease from aspergillus saitoi. | | 1972 | 4664737 |
| alkylation of ribonuclease from aspergillus saitoi with iodoacetate and iodoacetamide. | | 1973 | 4720055 |
| production of a new type of acid carboxypeptidase of molds of the aspergillus niger group. | the ability of 88 fungi, which had been obtained as high-potency strains for acid proteinase production, to produce a new type of acid carboxypeptidase (having on optimal ph of about 3 for hydrolysis of benzyloxycarbonyl-glutamyltyrosine) in surface koji culture was determined. among the aspergilli, substantial amounts of this new acid carboxypeptidase were produced by aspergillus saitoi, a. usamii, a. awamori, a. inuii, and a. niger. maximum yields of acid carboxypeptidase per gram of substrate ... | 1973 | 4796163 |
| production and some properties of a new type of acid carboxypeptidase of penicillium molds. | among some 38 strains of the genus penicillium we investigated seven wild-type strains (p. daleae ifo-6087, p. frequentans ahu-8328, p. funiculosum iam-7013, p. janthinellum ifo-8070, iam-7026, p. lividum iam-7200, and p. oxalicum ahu-8336) that were found to be excellent strains for a new type of acid carboxypeptidase production in a surface koji culture at 25 c. the production of acid carboxypeptidase was determined in various culture conditions in a koji culture. the maximum yields of acid ca ... | 1974 | 4833287 |
| photooxidation of ribonuclease from aspergillus saitoi. | | 1969 | 5354030 |
| some chemical and physical properties of ribonuclease from aspergillus saitoi. | | 1971 | 5577152 |
| interaction of ribonuclease from aspergillus saitoi with aristeromycin and its phosphate esters. | | 1971 | 5577154 |
| isolation and properties of a ribonuclease from aspergillus saitoi. | | 1967 | 5587601 |
| substrate specificity of ribonuclease from aspergillus saitoi. | | 1968 | 5709261 |
| inhibition of ribonuclease from aspergillus saitoi by nucleosides. | | 1969 | 5811789 |
| molecular weight and amino acid composition of acid proteinase of aspergillus saitoi. | | 1965 | 5865163 |
| molecular weight of acid proteinase of aspergillus saitoi. | | 1965 | 5886141 |
| n- and c-terminal residues in the acid proteinase of aspergillus saitoi. | | 1966 | 5943597 |
| melanins and resistance of fungi to lysis. | hyphal walls of aspergillus phoenicis and sclerotium rolfsii are composed of large amounts of glucose- and n-acetylhexosamine-containing polysaccharides, and the walls are extensively digested by streptomycete culture filtrates or by a mixture of purified chitinase and beta-(1 --> 3) glucanase preparations with the release of the monomeric units. a. phoenicis conidial walls also contain polymers of glucose and n-acetylhexosamine, but these walls are resistant to digestion by microorganisms or th ... | 1967 | 6032507 |
| carboxamidomethylation of a ribonuclease from aspergillus saitoi. | 1) the inactivation of a rnase from aspergillus saitoi (rnase ms) was studied to obtain information on its active site. 2) inactivation of rnase ms by iodoacetamide was greater at an alkaline ph, and was protected more by 2',(3')-amp than by 2',(3')-gmp. 3) analysis of the hydrolysis products with 6 n hcl and alkaline treatment of carboxamidomethylated rnase ms showed that the sites of reaction were one carboxyl group and one histidine residue. 4) since the incorporation of a carboxamidomethyl g ... | 1980 | 6257663 |
| characterization of two forms of base non-specific and adenylic acid preferential ribonuclease from aspergillus saitoi. | two forms of rnases (rnase ml and rnase mm) from aspergillus saitoi which are base non-specific and adenylic acid preferential were separated from each other by deae-cellulose column chromatography. they are indistinguishable with respect to enzymatic properties such as base preferability, ph optimum, kinetic constants measured with 2',3'-cump and 2',3'-ccmp as substrates, and effects of ionic strength, physical properties such as heat stability, isoelectric point and circular dichroism spectra, ... | 1983 | 6417118 |
| modification of a major ribonuclease from aspergillus saitoi with 1-cyclohexyl-3-(2-morpholinyl-(4)-ethyl)carbodiimide. | in order to investigate the role of carboxyl groups of a base non-specific ribonuclease from aspergillus saitoi [ec 3.1.27.1] (rnase m, molecular weight 36,000), the modification of rnase m with a water-soluble carbodiimide, 1-cyclohexyl-3-(2-morpholinyl-(4)-ethyl)carbodiimide(cmc), was studied. the inactivation of rnase m proceeded almost linearly with the incorporation of about 9.5 cmc moieties. the peptide backbone structure of the modified rnase m was practically the same as that of the nati ... | 1983 | 6578211 |
| purification and characterization of a glucoamylase from aspergillus saitoi. | 1. a major glucoamylase [ec 3.2.1.3] of aspergillus saitoi was purified by ultrafiltration followed by successive chromatography on deae-sephadex, ultrogel aca 44 and sp-sephadex. the purification achieved was 23-fold from crude extract with a yield of 21%. the purified enzyme, named gluc m1, was proved homogeneous as judged by polyacrylamide gel electrophoresis, isoelectric focusing, ultracentrifugation, and also from the absence of the glycosidase activities detected in crude extract. 2. gluc ... | 1981 | 6783630 |
| purification and characterization of a minor glucoamylase from aspergillus saitoi. | | 1981 | 6796572 |
| modification of a glucoamylase from aspergillus saitoi with 1-cyclohexyl-3-(2-morpholinyl-(4)-ethyl)carbodiimide. | 1. in order to elucidate the structure-function relation of a glucoamylase [ec 3.2.1.3, alpha-d-(1 leads to 4)-glucan glucohydrolase] from aspergillus saitoi (gluc m1), the reaction of gluc m1 with water-soluble carbodiimides was studied. 2. gluc m1 was inactivated most effectively by 1-cyclohexyl-3-(2-morpholinyl-(4)-ethyl)carbodiimide (cmc) at ph 4.5. 3. inactivation of gluc m1 with [14c]cmc proceeded with the incorporation of about 12 cmc moieties. from the results of amino acid analysis, tit ... | 1982 | 6802809 |
| n-bromosuccinimide oxidation of a glucoamylase from aspergillus saitoi. | 1. in order to elucidate the structure-function relation of a glucoamylase [ec 3.2.1.3, alpha-d-(1 leads to 4) glucan glucohydrolase] from aspergillus saitoi (gluc m1), the reaction of gluc m1 with nbs was studied. 2. the tryptophan residues in glu m1 were oxidized at various nbs/gluc m1 ratios. the enzymatic activity decreased to about 80% of that of the native gluc m1 with the oxidation of the first 2 tryptophan residues. the oxidation of these 2 tryptophan residues occurred within 0.2-0.5 s. ... | 1982 | 6807973 |
| steroid modifications with immobilized biocatalysts--use of immobilized enzyme-requiring cofactor regeneration and of immobilized mycelium. | two biological approaches have been investigated for specific modifications of steroids. the first one uses the purified enzyme for the specific dehydrogenation of androsterone to androstanedione. the enzyme used is 3 alpha-hydroxysteroid dehydrogenase which requires a cofactor (nad). a cofactor regeneration is needed so that the process could work continuously. the conjugation of two points (immobilization of the enzyme and optimization of the ratio methanol-water) allows a continuous work of t ... | 1982 | 6962578 |
| purification of an acidic alpha-d-mannosidase from aspergillus saitoi and specific cleavage of 1,2-alpha-d-mannosidic linkage in yeast mannan. | an acidic alpha-d-mannosidase (alpha-d-mannoside mannohydrolase, ec 3.2.1.24) has been isolated from culture filtrate of aspergillus saitoi. the extracellular alpha-mannosidase was homogeneous in polyacrylamide gel electrophoresis. the molecular weight of the enzyme was 51 000 and the isoelectric point ph 4.5. the purified enzyme has a ph optimum of 5.0, a km of 0.45 mm with baker's yeast mannan and has no activity towards p-nitrophenyl-alpha-d-mannoside. the mode of action of the enzyme has be ... | 1981 | 7011404 |
| primary structure of a minor ribonuclease from aspergillus saitoi. | 1. rnase ms, a base non-specific rnase from aspergillus saitoi was reduced and carboxymethylated (rcm-rnase ms). rcm-rnase ms was hydrolyzed with trypsin, and the trypsin digests were then treated with chymotrypsin. trypsin digests were also treated with staphylococcus protease and with chymotrypsin, separately. 2. by the analyses of the amino acid sequences of the peptides formed, the alignment of these peptides in rcm-rnase ms was determined. 3. from the digest of heat-denatured rnase ms with ... | 1982 | 7096302 |
| an alpha-mannosidase purified from aspergillus saitoi is specific for alpha 1,2 linkages. | | 1980 | 7437073 |
| effect of irreversibility on the thermodynamic characterization of the thermal denaturation of aspergillus saitoi acid proteinase. | the thermal denaturation of the acid proteinase from aspergillus saitoi was studied by cd and differential scanning calorimetry (dsc). this process seemed to be completely irreversible, as protein samples that were heated to temperatures at which the transition had been completed and then cooled at 25 degrees c did not show any reversal of the change in the cd signal. similar results were obtained with dsc. nevertheless, we were able to detect the presence of reversibly unfolded species in exper ... | 1995 | 7487958 |
| molecular cloning of the cdna and gene for an elastinolytic aspartic proteinase from aspergillus fumigatus and evidence of its secretion by the fungus during invasion of the host lung. | hydrolysis of structural proteins in the lung by extracellular proteinases secreted by aspergillus fumigatus is thought to play a significant role in invasive aspergillosis. this fungus was found previously to secrete an elastinolytic serine proteinase and a metalloproteinase. we report that a. fumigatus also secretes an aspartic proteinase (aspergillopepsin f) that can catalyze hydrolysis of the major structural proteins of basement membrane, elastin, collagen, and laminin. the ph optimum for t ... | 1995 | 7558282 |
| the carbohydrate moiety of the acid carboxypeptidase from aspergillus saitoi. | acid carboxypeptidase from aspergillus saitoi is a glycoprotein that contains both n- and o-linked sugar chains. the n-glycanase released high-mannose type oligosaccharides that were separated into eight components on hplc. one, which had a unique structure of man11glcnac2, was characterized. mild alkali treatment of the carboxypeptidase, under conditions that effect beta-elimination, yielded d-mannose. deglycosylation of the carboxypeptidase with endo-beta-n-acetylglucosaminidase and alpha-mann ... | 1993 | 7764137 |
| cloning and expression of the carboxypeptidase gene from aspergillus saitoi and determination of the catalytic residues by site-directed mutagenesis. | carboxypeptidase from aspergillus saitoi removes acidic, neutral and basic amino acids as well as proline from the c-terminal position at ph 2-5. cpds, a cdna encoding a. saitoi carboxypeptidase, was cloned and expressed. analysis of the 1816-nucleotide sequence revealed a single open reading frame coding for 523 amino acids. when a. saitoi carboxypeptidase cdna was expressed in yeast cells, carboxypeptidase activity was detected in the cell extract and was immunostained with a 72 kda protein wi ... | 1995 | 7772020 |
| crystal structure of ribonuclease ms (as a ribonuclease t1 homologue) complexed with a guanylyl-3',5'-cytidine analogue. | a ribonuclease t1 homologue, ribonuclease ms (rnase ms) from aspergillus saitoi, has been crystallized as a complex with a substrate analogue gfpc where the 2'-hydroxyl (2'-oh) group of guanosine in guanylyl-3',5'-cytidine (gpc) is replaced by the 2'-fluorine (2'-f) atom to prevent transesterification. the crystal structure of the complex was solved at 1.8-a resolution to a final r-factor of 0.204. the role of his92 (rnase t1 numbering) as the general acid catalyst was confirmed. of the two alte ... | 1993 | 8218254 |
| molecular cloning and nucleotide sequence of the 1,2-alpha-d-mannosidase gene, msds, from aspergillus saitoi and expression of the gene in yeast cells. | a full-length cdna encoding 1,2-alpha-d-mannosidase (ec 3.2.1.113) from aspergillus saitoi was cloned. analysis of the 1718 bp nucleotide sequence of the cdna revealed a single open reading frame with 1539 nucleotides of 1,2-alpha-d-mannosidase gene, msds. the predicted amino-acid sequence of 1,2-alpha-d-mannosidase consists of 513 residues with a molecular mass of 55,767 and is 70%, 26% and 35% identity with those of penicillium citrinum 1,2-alpha-d-mannosidase, yeast alpha-mannosidase, and mou ... | 1995 | 8519794 |
| separation of biosynthetic oligosaccharide branch isomers using high-performance liquid chromatography on a porous two-dimensional graphite stationary phase. | oligomannosidic branch isomers (structures differing only in the branch location of a single residue) which are biosynthetic intermediates in yeast and higher eukaryotics have been separated using high-performance liquid chromatography (hplc) on porous graphatized carbon (pgc) columns, a stationary phase of two-dimensional crystalline carbon. a mixture of two man6glcnac isomers from igm, which was determined from 1h nmr analysis, was completely separated by pgc-hplc. mixtures of larger yeast oli ... | 1996 | 8954551 |
| characterization of the s1 subsite specificity of aspergillopepsin i by site-directed mutagenesis. | the structural determinants of s1 substrate specificity of aspergillopepsin i (api; ec 3.4.23.18), an aspartic proteinase from aspergillus saitoi, were investigated by site-directed mutagenesis. aspartic proteinases generally favor hydrophobic amino acids at p1 and p1'. however, api accommodates a lys residue at p1, which leads to activation of trypsinogen. on the basis of amino acid sequence alignments of aspartic proteinases, asp-76 and ser-78 of api are conserved only in fungal enzymes with t ... | 1996 | 8982865 |
| hydrophobic mannosides act as acceptors for trypanosome alpha-mannosyltransferases. | a series of hydrophobic mannosides were synthesized and tested for their ability to act as acceptor substrates for mannosyltransferases in a trypanosoma brucei cell-free system. the thiooctyl alpha-mannosides and octyl alpha-mannosides all accepted single mannose residues in alpha-linkage, as judged by thin layer chromatography of the products before and after jack bean alpha-mannosidase digestion. the mannosylation reactions were inhibited by amphomycin, suggesting that the immediate donor was ... | 1997 | 9184836 |
| identification of the glycosylation site and glycan structures of recombinant endopolygalacturonase ii by mass spectrometry. | a series of mass spectrometric experiments was performed to characterize the carbohydrate chains attached to endopolygalacturonase ii (epg-ii) overexpressed in aspergillus niger. first, an aliquot of trypsin-digested epg-ii was analyzed by capillary high-performance liquid chromatography (hplc) coupled with electrospray ionization (esi) mass spectrometry (hplc/ms). the esi mass spectrometer was operated in the tandem mass spectrometric (ms/ms) mode and utilized precursor ion scanning of the hexn ... | 1997 | 9276972 |
| five crucial carboxyl residues of 1,2-alpha-mannosidase from aspergillus saitoi (a. phoenicis), a food microorganism, are identified by site-directed mutagenesis. | an acidic 1,2-alpha-mannosidase from fungus, aspergillus saitoi (now designated aspergillus phoenicis), is highly specific for 1,2-alpha-mannosidic linkage in the high-mannose type oligosaccharide at ph 5.0. the predicted amino acid sequence of several peptide regions, including aspartic acid and glutamic acid, bears striking similarities to 1,2-alpha-mannosidases from fungi, yeast and mouse. active site determination of the enzyme expressed in saccharomyces cerevisiae cells was performed by sit ... | 1997 | 9325167 |
| [structures and functions of ribonucleases]. | 1. in order to understand the differences in ph optima and reaction rates of rnase a towards low molecular weight substrates and polymer substrates, the subsite structure of bovine pancreatic rnase a was studied. the kinetic studies of various sizes of oligouridylic acids showed that the size of the subsite is three nucleotides long. the kinetic studies on the inhibition of pup, x-ray crystallographies of rnase a-apc and ptp complexes, 31p-nmr studies on the binding of rnase a-pap, and ptp showe ... | 1997 | 9357326 |
| in vitro conversion of the carbohydrate moiety of fungal glycoproteins to mammalian-type oligosaccharides--evidence for n-acetylglucosaminyltransferase-i-accepting glycans from trichoderma reesei. | to investigate the potential of filamentous fungi to synthesize n-glycans that are convertible to a mammalian type, in vitro glycosylation assays were performed. recombinant human n-acetylglucosaminyltransferase i, human beta-1,4-galactosyltransferase and rat alpha-2,6-sialyltransferase were successively used to mimic part of the mammalian glycosylation synthesis pathway. high-mannose carbohydrates on trichoderma reesei cellobiohydrolase i were converted to a hybrid mammalian-type structure. suc ... | 1997 | 9395316 |
| use of hemicellulose hydrolysate for beta-glucosidase fermentation. | hydrolysis of cellulose by trichoderma cellulases often results in a mixture of glucose, cellobiose, and low-mol-wt cellodextrins. cellobiose is nonfermentable for most yeasts, and therefore it has to be hydrolyzed to glucose by beta-glucosidase prior to ethanol fermentation. in the present study, the beta-glucosidase production of one penicillium and three aspergillus strains, which were previously selected out of 24 strains, was investigated on steam pretreated willow. both steam-pretreated wi ... | 1998 | 9627384 |
| molecular and enzymic properties of recombinant 1, 2-alpha-mannosidase from aspergillus saitoi overexpressed in aspergillus oryzae cells. | for the construction of an overexpression system of the intracellular 1,2-alpha-mannosidase (ec 3.2.1.113) gene (msds) from aspergillus saitoi (now designated aspergillus phoenicis), the n-terminal signal sequence of the gene was replaced with that of the aspergillopepsin i (ec 3.4.23.18) gene (apns) signal, one of the same strains as described previously. then the fused 1, 2-alpha-mannosidase gene (f-msds) was inserted into the noti site between p-no8142 and t-agda in the plasmid pnan 8142 (9.5 ... | 1999 | 10215597 |
| molecular cloning and enzymatic characterization of a trichoderma reesei 1,2-alpha-d-mannosidase. | a cdna encoding 1,2-alpha-d-mannosidase mds 1 from trichoderma reesei was cloned. the largest open reading frame occupied 1571 bp. the predicted sequence contains 523 amino acid residues for a calculated molecular mass of 56,266 da and shows high similarity to the amino acid sequences of 1,2-alpha-d-mannosidases from aspergillus saitoi and penicillium citrinum (51.6 and 51.0% identity, respectively). t. reesei mannosidase was produced as a recombinant enzyme in the yeast pichia pastoris. replace ... | 2000 | 10682284 |
| fabrication, characterization, and enzymatic activity of encapsulated fungal protease--fatty lipid biocomposite films. | encapsulation of an aspartic protease from the fungus aspergillus saitoi (f-prot) in thermally evaporated fatty acid films by a simple beaker-based immersion technique under enzyme-friendly conditions is described. the approach is based on diffusion of the enzyme from aqueous solution, driven primarily by attractive electrostatic interaction between charged groups on the enzyme surface and ionized lipid molecules in the film. the encapsulated enzyme molecules could be "pumped out" of the biocomp ... | 2000 | 11008764 |
| structure of aspergillopepsin i from aspergillus phoenicis: variations of the s1'-s2 subsite in aspartic proteinases. | the crystal structure of aspergillopepsin i (ap) from aspergillus phoenicis has been determined at 2.18 a resolution and refined to r and r(free) factors of 21.5 and 26.0%, respectively. ap has the typical two beta-barrel domain structure of aspartic proteinases. the structures of the two independent molecules are partly different, exemplifying the flexible nature of the aspartic proteinase structure. notably, the 'flap' in one molecule is closer, with a largest separation of 4.0 a, to the activ ... | 2001 | 11418762 |
| purification and properties of a thermostable extracellular beta-d-xylosidase produced by a thermotolerant aspergillus phoenicis. | a beta-d-xylosidase was purified from cultures of a thermotolerant strain of aspergillus phoenicis grown on xylan at 45 degrees c. the enzyme was purified to homogeneity by chromatography on deae-cellulose and sephadex g-100. the purified enzyme was a monomer of molecular mass 132 kda by gel filtration and sds-page. treatment with endoglycosidase h resulted in a protein with a molecular mass of 104 kda. the enzyme was a glycoprotein with 43.5% carbohydrate content and exhibited a pl of 3.7. opti ... | 2001 | 11420656 |
| identification, classification and phylogeny of the aspergillus section nigri inferred from mitochondrial cytochrome b gene. | the partial mitochondrial cytochrome b gene from 32 strains of 12 species belonging to aspergillus section nigri was amplified by the polymerase chain reaction and sequenced directly. using 402 nucleotide characters, nucleotide-based and amino acid-based phylogenetic trees were inferred and the genetic divergence among the species was evaluated. based on analyses of the 402-bp nucleotide and 133-amino acid sequences, strains were divided into 11 dna types and five amino acid types. aspergillus n ... | 2001 | 11425482 |
| structural and mechanistic insight into the inhibition of aspartic proteases by a slow-tight binding inhibitor from an extremophilic bacillus sp.: correlation of the kinetic parameters with the inhibitor induced conformational changes. | we present here the first report of a hydrophilic peptidic inhibitor, atbi, from an extremophilic bacillus sp. exhibiting a two-step inhibition mechanism against the aspartic proteases, pepsin and f-prot from aspergillus saitoi. kinetic analysis shows that these proteases are competitively inhibited by atbi. the progress curves are time-dependent and consistent with slow-tight binding inhibition: e + i right arrow over left arrow (k(3), k(4)) ei right arrow over left arrow (k(5), k(6)) ei. the k ... | 2001 | 11560501 |
| on the preparation, characterization, and enzymatic activity of fungal protease-gold colloid bioconjugates. | we present herein details pertaining to the preparation of bioconjugates of colloidal gold with aspartic protease from the fungus aspergillus saitoi (f-prot) and their characterization and enzymatic activity. simple mixing of the colloidal gold and protein solutions under protein-friendly conditions (ph = 3) followed by centrifugation (to remove uncomplexed gold nanoparticles and protein molecules) results in the formation of the fungal protease-gold nanoparticle conjugates. the protein-gold nan ... | 2001 | 11562186 |
| the broadly neutralizing anti-human immunodeficiency virus type 1 antibody 2g12 recognizes a cluster of alpha1-->2 mannose residues on the outer face of gp120. | 2g12 is a broadly neutralizing human monoclonal antibody against human immunodeficiency virus type-1 (hiv-1) that has previously been shown to bind to a carbohydrate-dependent epitope on gp120. here, site-directed mutagenesis and carbohydrate analysis were used to define further the 2g12 epitope. extensive alanine scanning mutagenesis showed that elimination of the n-linked carbohydrate attachment sequences associated with residues n295, n332, n339, n386, and n392 by n-->a substitution produced ... | 2002 | 12072529 |
| cloning and expression of the exo-beta-d-1,3-glucanase gene (exgs) from aspergillus saitoi. | a gene of exo-1,3-beta-d-glucanase (exgs) was cloned from a koji mold, aspergillus saitoi, genomic dna using pcr. the exgs has an orf comprising 2832 bp, which contains one intron of 45 bp, and encodes 945 amino acids. the deduced amino acid sequences showed that the exgs has a non-homologous linker region consisting of 180 amino acids, which encompassed highly conserved regions observed in exg homologues from filamentous fungi. a recombinant protein (exgs) has been recovered from the cultural f ... | 2002 | 12224649 |
| structural determination of the n-glycans of a lepidopteran arylphorin reveals the presence of a monoglucosylated oligosaccharide in the storage protein. | the structures of the oligosaccharides attached to arylphorin from chinese oak silkworm, antheraea pernyi, have been determined. arylphorin, a storage protein present in fifth larval hemolymph, contained 4.8% (w/w) of carbohydrate that was composed of fuc:glcnac:glc:man=0.2:4.0:1.4:13.6 moles per mole protein. four moles of glcnac in oligomannose-type oligosaccharides strongly suggest that the protein contains two n-glycosylation sites. normal-phase hplc and mass spectrometry oligosaccharide pro ... | 2003 | 12626419 |
| perturbation of the wing color pattern of a swallowtail butterfly, papilio xuthus, induced by acid carboxypeptidase. | hypodermal injection of toughmac-e, a digestive mixture composed of nine digestive components, or molsin induced perturbation of the wing color pattern in 0- to 2-day-old pupae of papilio xuthus, but had no effect on prepupae or 3- to 4-day-old pupae. the effective component in toughmac-e was identified as molsin, an acid carboxypeptidase of aspergillus saitoi which specifically liberates tyrosine and phenylalanine from the c-terminal residues of proteins. the pattern perturbation occurred in ei ... | 2003 | 12692391 |
| identification of catalytic residues of ca2+-independent 1,2-alpha-d-mannosidase from aspergillus saitoi by site-directed mutagenesis. | the roles of six conserved active carboxylic acids in the catalytic mechanism of aspergillus saitoi 1,2-alpha-d-mannosidase were studied by site-directed mutagenesis and kinetic analyses. we estimate that glu-124 is a catalytic residue based on the drastic decrease of kcat values of the e124q and e124d mutant enzyme. glu-124 may work as an acid catalyst, since the ph dependence of its mutants affected the basic limb. d269n and e411q were catalytically inactive, while d269e and e411d showed consi ... | 2003 | 12702721 |
| synthesis of fimh receptor-active manno-oligosaccharides by reverse hydrolysis using alpha-mannosidases from penicillium citrinum, aspergillus phoenicis and almond. | recombinant penicillium citrinum alpha-1,2-mannosidase, expressed in aspergillus oryzae, was employed to carry out regioselective synthesis of alpha- d-mannopyranosyl-(1-->2)- d-mannose. yields (w/w) of 16.68% disaccharide, 3.07% trisaccharide and 0.48% tetrasaccharide were obtained, with alpha1-->2 linkages present at 98.5% of the total linkages formed. non-specific alpha-mannosidase from almond was highly efficient in reverse hydrolysis and oligosaccharide yields of 45-50% were achieved. the p ... | 2004 | 12910329 |
| new potent antioxidative hydroxyflavanones produced with aspergillus saitoi from flavanone glycoside in citrus fruit. | potent antioxidative hydroxyflavanones were produced with aspergillus saitoi from hesperidin or naringin, which are flavanone glycosides in citrus fruit with weak antioxidative activity. the hydroxyflavanone produced from hesperidin was identified as 8-hydroxyhesperetin (8-hhe), a novel substance, and those from naringin were identified as carthamidin (6-hydroxynaringenin) and isocarthamidin (8-hydroxynaringenin) by fab-ms, 1h-nmr and 13c-nmr analyses. the antioxidative activity of these hydroxy ... | 2003 | 12913285 |
| [on the activation of bovine trypsinogen by a crystallized protease from aspergillus saitoi]. | | 1960 | 13702766 |
| microbiological production ofgluconic acid by egyptian mould fungi. (iii). production of gluconic acid by aspergillus phoenicis in submerged cultures. | | 1960 | 13836595 |
| chromatographic purification and physical homogeneity of acid proteinase of aspergillus saitoi. | | 1965 | 14336073 |
| use of different proteases working in acidic conditions to improve sequence coverage and resolution in hydrogen/deuterium exchange of large proteins. | the combination of hydrogen exchange and mass spectrometry has been widely used in structural biology, providing views on protein structure and protein dynamics. one of the constraints is to use proteases working at low ph and low temperature to limit back-exchange during proteolysis. although pepsin works in these conditions and is currently used in such experiments, sequence coverage is not always complete especially for large proteins, and the spatial resolution of the exchange rate is limite ... | 2003 | 14587084 |
| hydrolytic cleavage of purine ribonucleosides in aspergillus phoenicis. | cell-free extracts of nitrate-grown aspergillus phoenicis could catalyze the hydrolytic cleavage of the n-glycosidic bond of inosine, guanosine and adenosine to the corresponding base and ribose by the nucleoside hydrolase. no evidence was obtained concerning the hydrolytic degradation of n-glycosidic bond of pyrimidine ribonucleosides namely cytidine and uridine by the same extracts. optimum ph and temperature for adenosine, guanosine and inosine hydrolysis were the same at ph 3.5 and 55 degree ... | 2003 | 14625894 |
| influence of temperature on the properties of the xylanolytic enzymes of the thermotolerant fungus aspergillus phoenicis. | this study reports on the effects of growth temperature on the secretion and some properties of the xylanase and beta-xylosidase activities produced by a thermotolerant aspergillus phoenicis. marked differences were observed when the organism was grown on xylan-supplemented medium at 25 degrees c or 42 degrees c. production of xylanolytic enzymes reached maximum levels after 72 h of growth at 42 degrees c; and levels were three- to five-fold higher than at 25 degrees c. secretion of xylanase and ... | 2004 | 14767676 |
| the production, purification and characterisation of two novel alpha-d-mannosidases from aspergillus phoenicis. | 1,6-alpha-d-mannosidase from aspergillus phoenicis was purified by anion-exchange chromatography, chromatofocussing and size-exclusion chromatography. the apparent molecular weight was 74 kda by sds-page and 81 kda by native-page. the isoelectric point was 4.6. 1,6-alpha-d-mannosidase had a temperature optimum of 60 degrees c, a ph optimum of 4.0-4.5, a k(m) of 14 mm with alpha-d-manp-(1-->6)-d-manp as substrate. it was strongly inhibited by mn(2+) and did not need ca(2+) or any other metal cofa ... | 2005 | 15721331 |
| production of cellulase/beta-glucosidase by the mixed fungi culture of trichoderma reesei and aspergillus phoenicis on dairy manure. | a cellulase production process was developed by growing the fungi trichoderma reesei and aspergillus phoenicis on dairy manure. t. reesei produced a high total cellulase titer (1.7 filter paper units [fpu]/ml, filter paper activity) in medium containing 10 g/l of manure (dry basis [w/w]), 2 g/l kh2po4, 2 ml/l of tween-80, and 2mg/l of cocl2. however, beta-glucosidase activity in the t. reesei-enzyme system was very low. t. reesei was then cocultured with a. phoenicis to enhance the beta-glucosid ... | 2005 | 15917591 |
| xylanase production by fungal strains on spent sulphite liquor. | xylanase production by seven fungal strains was investigated using concentrated spent sulphite liquor (sslc), xylan and d: -xylose as carbon substrates. an sslc-based medium induced xylanase production at varying levels in all of these strains, with aspergillus oryzae nrrl 3485 and aspergillus phoenicis atcc 13157 yielding activities of 164 and 146 u ml(-1), respectively; these values were higher than those obtained on xylan or d: -xylose with the same fungal strains. the highest xylanase activi ... | 2005 | 15944854 |
| local dynamics measured by hydrogen/deuterium exchange and mass spectrometry of creatine kinase digested by two proteases. | hydrogen/deuterium exchange coupled to mass spectrometry has been used to investigate the structure and dynamics of native dimeric cytosolic muscle creatine kinase. the protein was incubated in d2o for various time. after h/d exchange and rapid quenching of the reaction, the partially deuterated protein was cleaved in parallel by two different proteases (pepsin or type xiii protease from aspergillus saitoi) to increase the sequence coverage and spatial resolution of deuterium incorporation. the ... | 2005 | 16023284 |
| a single free cysteine residue and disulfide bond contribute to the thermostability of aspergillus saitoi 1,2-alpha-mannosidase. | aspergillus saitoi 1,2-alpha-mannosidase contains three conserved cysteine residues (cys334, cys363, and cys443). we showed that cys334 and cys363 are involved in a disulfide bond, and that cys443 contains a free thiol group. the cysteines were not essential for the activity analyzed by site-directed mutagenesis and kinetics. the substitution at each cysteine residue greatly destabilized the enzyme. the t(m) values of wt, c443a, c443g, c443s, and c443t were 55.8, 51.9, 50.2, 50.0, and 52.8 degre ... | 2005 | 16306691 |
| functional characterization of the hansenula polymorpha hoc1, och1, and ocr1 genes as members of the yeast och1 mannosyltransferase family involved in protein glycosylation. | the alpha-1,6-mannosyltransferase encoded by saccharomyces cerevisiae och1 (scoch1) is responsible for the outer chain initiation of n-linked oligosaccharides. to identify the genes involved in the first step of outer chain biosynthesis in the methylotrophic yeast hansenula polymorpha, we undertook the functional analysis of three h. polymorpha genes, hphoc1, hpoch1, and hpocr1, that belong to the och1 family containing seven members with significant sequence identities to scoch1. the deletions ... | 2006 | 16407250 |