novel easily accessible glucosidase inhibitors: 4-hydroxy-5-alkoxy-1,2-cyclohexanedicarboxylic acids.glycosidases are very important enzymes involved in a variety of biochemical processes with a special importance to biotechnology, food industry, and pharmacology. novel structurally simple inhibitors derived from cyclohexane-1,2-dicarboxylic acids were synthesized and tested against several fungal glycosidases from aspergillus oryzae and penicilliumcanescens. the presence of at least two carboxylic groups and one hydroxy group was essential for efficient inhibition. significant selective inhibi ...200919084826
mathematical modeling of the development of antioxidant activity in soybeans fermented with aspergillus oryzae and aspergillus awamori in the solid state.the kinetics of the development fungal growth, beta-glucosidase activity, total phenolics, and dpph scavenging in soybeans fermented with aspergillus oryzae and aspergillus awamori were studied over an 8-day incubation period. modified logistic equations were then used to describe the experimental fermentation profiles. in all cases the models were consistent and the parametric estimations were statistically significant. the predicted values have high coefficients of linear correlation with the ...200919099459
cloning and heterologous transcription of a polyketide synthase gene from the lichen solorina crocea.lichens and most ascomycete fungi produce polyketide secondary metabolites often with valuable biological activities. their biosynthesis is primarily governed by large iterative multifunctional type i polyketide synthases. although there has been good progress studying filamentous non-lichenized fungi, there is limited information on polyketide biosynthesis in lichens and their mycobionts, due to their slow growth, difficulties in establishing pure cultures, and the absence of methods for direct ...200919100326
construction of quintuple protease gene disruptant for heterologous protein production in aspergillus oryzae.aspergillus oryzae has received attention as a host for heterologous protein production. however, a. oryzae has 134 protease genes, which is recognized to be one of the major reasons for the proteolytic degradation of heterologously produced proteins. we previously reported that double disruption of the protease genes (tppa and pepe) improved heterologous protein (human lysozyme) production by a. oryzae. in this study, we performed successive round of five protease genes (tppa, pepe, nptb, dppiv ...200919107471
comparative analysis of oligosaccharide specificities of fucose-specific lectins from aspergillus oryzae and aleuria aurantia using frontal affinity chromatography.aleuria aurantia lectin (aal) is widely used to estimate the extent of alpha1,6-fucosylated oligosaccharides and to fractionate glycoproteins for the detection of specific biomarkers for developmental antigens. our previous studies have shown that aspergillus oryzae lectin (aol) reflects the extent of alpha1,6-fucosylation more clearly than aal. however, the subtle specificities of these lectins to fucose linked to oligosaccharides through the 2-, 3-, 4-, or 6-position remain unclear, because la ...200919109923
construction of a citrinin gene cluster expression system in heterologous aspergillus oryzae.filamentous fungi are considered an attractive resource for the discovery and production of bioactive compounds. to facilitate molecular breeding, biosynthetic genes must be rapidly identified. but, even after the chemical structure of a compound is identified, finding the corresponding biosynthetic genes in the fungal genome still remains a challenge. in an attempt to overcome this difficulty and to easily characterize each gene in a cluster, we constructed a heterologous expression system usin ...200819111642
isolation of industrial strains of aspergillus oryzae lacking ferrichrysin by disruption of the dffa gene.based on studies using laboratory strains, the efficiency of gene disruption in aspergillus oryzae, commonly known as koji mold, is low; thus, gene disruption has rarely been applied to the breeding of koji mold. to evaluate the efficiency of gene disruption in industrial strains of a. oryzae, we produced ferrichrysin biosynthesis gene (dffa) disruptants using three different industrial strains as hosts. pcr analysis of 438 pyrithiamine-resistant transformants showed dffa gene disruption efficie ...200819111645
use of saccharum spontaneum (wild sugarcane) as biomaterial for cell immobilization and modulated ethanol production by thermotolerant saccharomyces cerevisiae vs3.saccharum spontaneum is a wasteland weed consists of 45.10+/-0.35% cellulose and 22.75+/-0.28% of hemicellulose on dry solid (ds) basis. aqueous ammonia delignified s. spontaneum yielded total reducing sugars, 53.91+/-0.44 g/l (539.10+/-0.55 mg/g of substrate) with a hydrolytic efficiency of 77.85+/-0.45%. the enzymes required for hydrolysis were prepared from culture supernatants of aspergillus oryzae mtcc 1846. a maximum of 0.85+/-0.07 iu/ml of filter paperase (fpase), 1.25+/-0.04 iu/ml of car ...200919114303
functional expression of miraculin, a taste-modifying protein in escherichia coli.miraculin isolated from red berries of richadella dulcifica, a native shrub of west africa, has the unusual property of modifying a sour taste into a sweet one. this homodimer protein consists of two glycosylated polypeptides that are cross-linked by a disulfide bond. recently, functional expression of miraculin was reported in host cells with the ability to glycosylate proteins, such as lettuce, tomato and the microbe aspergillus oryzae, but not escherichia coli. thus, a question remains as to ...200919122203
analysis of extracellular proteins of aspergillus oryzae grown on soy sauce koji.aspergillus oryzae as 3.951 is widely used in chinese soy sauce manufacture, but little is known about the profiles of the extracellular proteins from the culture of soybean koji. in this study, we carried out maldi-tof/tof ms analysis of extracellular proteins during koji culture. besides well-known proteins (taa and oryzin), a variety of aminopeptidase and proteases were identical at the proteome level. this suggests that a. oryzae as 3.951 has a powerful capacity to digest soybean protein.200919129648
alternative processing of proproteins in aspergilli kexb gene disruptants under hyperosmotic conditions.disruption of the kexb gene encoding a subtilisin-like processing protease in aspergillus oryzae and aspergillus nidulans led to remarkable morphological defects, and these phenotypes were suppressed under hyperosmotic conditions. in this study, we investigated to determine whether non-kexb proteases might complement the in vivo function of kexb in the two aspergillus kexb disruptants. neither overexpression of opsa or opsb encoding a. oryzae aspartyl proteases homologous to yeast yapsins (yps1/ ...200919129662
disruption of the aopex11-1 gene involved in peroxisome proliferation leads to impaired woronin body formation in aspergillus oryzae.the woronin body, a unique organelle found in the pezizomycotina, plugs the septal pore upon hyphal damage to prevent excessive cytoplasmic bleeding. although it was previously shown that the woronin body buds out from the peroxisome, the relationship between peroxisomal proliferation/division and woronin body differentiation has not been extensively investigated. in this report, we examined whether pex11 required for peroxisomal proliferation participates in woronin body formation in aspergillu ...200919136573
chemopreventive effects of fermented brown rice and rice bran against 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone-induced lung tumorigenesis in female a/j mice.the most common cause of human lung cancer is suggested to be exposure to the carcinogens in tobacco smoke. among the multiple chemicals in tobacco smoke, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (nnk) has been regarded as one of the important agents for generation of lung cancers. previously, our studies proved that fermented brown rice and rice bran (fbra) has chemopreventive effects against carcinogenesis of the colon, liver, stomach, bladder and esophagus. in the present study, we exam ...200919148502
isolation of aspergillus oryzae mutants for heterologous protein production from a double proteinase gene disruptant.aspergillus oryzae has attracted much attention as a host for heterologous protein production because of its high secretion ability and safety. however, there have been only a few reports on construction of this organism to improve its properties as a production host. we previously reported that the double disruptant of the proteinase gene (tppa, pepe) improved human lysozyme (hly) production. in this double disruptant, however, the hly expression plasmid cannot be removed due to its random inte ...200919151971
codon optimization prevents premature polyadenylation of heterologously-expressed cellulases from termite-gut symbionts in aspergillus achieve high expression of glycoside hydrolase family 45 endoglucanase (rssym45eg1) from a symbiotic protist of the termite reticulitermes speratus, synthetic sequence rssym45eg1-co, in which the codon usage was adjusted to that of the highly-expressed tef1 gene encoding translation elongation factor 1alpha, was prepared and introduced into a. oryzae. the transcript level of rssym45eg1-co was 1.8-fold higher than that of rssym45eg1. in cells harboring rssym45eg1, but not rssym45eg1-co, trunca ...200819164877
natural killer cell activity, lymphocyte proliferation, and cytokine profile in tumor-bearing mice treated with mapa, a magnesium aggregated polymer from aspergillus oryzae.the present study examined the effects of mapa, an antitumor aggregated polymer of protein magnesium ammonium phospholinoleate-palmitoleate anhydride, isolated from aspergillus oryzae, on concanavalin a (con a)-induced spleen cell proliferation, cytokine production and on natural killer (nk) cell activity in ehrlich ascites tumor-bearing mice. the ehrlich ascites tumor (eat) growth led to diminished mitogen-induced expansion of spleen cell populations and total nk activity. this was accompanied ...200319180795
rapid screening and isolation of a fungus for sitosterol to androstenedione biotransformation.androstenedione (ad) is a steroid intermediate used in the pharmaceutical industry for the production of several important anabolic drugs. an important route for producing ad is by the microbial transformation of sterols. microbes capable of efficiently transforming sterols to ad are few and newer strains need to be isolated. conventional procedures for screening and isolation are time consuming. a new procedure was used for screening and isolation of fungal microorganisms capable of biotransfor ...200919189060
enzymatic hydrolysis of recovered protein from frozen small croaker and functional properties of its protein isolate were recovered from frozen small croaker using ph shift. the partial enzymatic hydrolysates were fractionated as soluble and insoluble parts. they were dried using the drum dryer and their functional properties were examined. the total nitrogen content of the enzymatic hydrolysates ranged from 12.9% to 13.7%. the degree of hydrolysis of precipitates was 18.2% and 12.2% for croaker hydrolysates treated with protamex 1.5 mg (bacilllus protease complex) and flavourzyme 500 mg ( ...200919200081
kinetic resolution of alpha-lipoic acid via enzymatic differentiation of a remote stereocenter.kinetic resolution of alpha-lipoic acid, a case of remote stereocenter discrimination, was accomplished using lipase from aspergillus oryzae wz007. performance of this lipase was investigated for enantioselective esterification of (s)-alpha-lipoic acid, leaving the target product (r)-alpha-lipoic acid in unreacted form. the effects of chain length of alcohol, type of solvent, molar ratio of alcohol:acid, and reaction temperature were studied. the optimum reaction conditions were found to be este ...200919212787
comparative analysis of core-fucose-binding lectins from lens culinaris and pisum sativum using frontal affinity chromatography.lens culinaris lectin (lca) is a useful probe for the detection in serum of a core-fucosylated alpha-fetoprotein, called afp-l3 fraction, which is a well-known marker for the diagnosis and prognosis of hepatocellular carcinoma. here we performed a systematic quantitative interaction analysis of lca and its close homolog, pisum sativum lectin (psa), by frontal affinity chromatography with 143 pyridylaminated (pa) glycans including a series of core-fucosylated glycans. both lectins showed binding ...200919218400
the dddp gene, encoding a novel enzyme that converts dimethylsulfoniopropionate into dimethyl sulfide, is widespread in ocean metagenomes and marine bacteria and also occurs in some ascomycete fungi.the marine alphaproteobacterium roseovarius nubinhibens ism can produce the gas dimethyl sulfide (dms) from dimethylsulfoniopropionate (dmsp), a widespread secondary metabolite that occurs in many phytoplankton. roseovarius possesses a novel gene, termed dddp, which when cloned, confers on escherichia coli the ability to produce dms. the dddp polypeptide is in the large family of m24 metallopeptidases and is wholly different from two other enzymes, dddd and dddl, which were previously shown to g ...200919220400
secretory expression of the non-secretory-type lentinula edodes laccase by aspergillus oryzae.the shiitake mushroom, lentinula edodes, has an extracelluar secretory-type laccase, lcc1, and a fruiting-body-accumulation-type laccase, lcc4. we previously reported the production of lcc1 by plant cells, but had difficulty producing lcc4. here, we report the production of lcc1 and lcc4 by aspergillus oryzae and the extracellular secretory production of lcc4 using a modified secretion signal peptide (sp) from lcc1. sp-lcc4 produced by a. oryzae had biochemical activities similar to lcc4 produce ...200919230633
enrichment of phenolics and free radical scavenging property of wheat koji prepared with two filamentous the present study, total phenolic content and antioxidant property of wheat (54% ethanolic extract) was drastically enhanced when fermented with two gras filamentous fungi, aspergillus oryzae and aspergillus awamori nakazawa. a. oryzae yielded significant (p<0.05) amounts of phenolic compounds, dpph* (2,2-diphenyl-1-picrylhydrazyl) and abts*(+) [2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid)] scavenging properties on 4th day of incubation compared to a. awamori on 5th day of incubatio ...200919232489
aspergillus flavus genomics as a tool for studying the mechanism of aflatoxin formation.aspergillus flavus is a weak pathogen that infects plants, animals and humans. when it infects agricultural crops, however, it produces one of the most potent carcinogens known (aflatoxins). to devise strategies to control aflatoxin contamination of pre-harvest agricultural crops and post-harvest grains during storage, we launched the a. flavus genomics program. the major objective of this program is the identification of genes involved in aflatoxin biosynthesis and regulation, as well as in pat ...200819238624
characterization of two distinct feruloyl esterases, aofaeb and aofaec, from aspergillus oryzae.two hypothetical proteins xp_001818628 and xp_001819091 (designated aofaeb and aofaec, respectively), showing sequence identity with known type-c feruloyl esterases, have been found in the genomic sequence of aspergillus oryzae. we cloned the putative a. oryzae feruloyl esterase-encoding genes and expressed them in pichia pastoris. both purified recombinant aofaeb (raofaeb) and aofaec (raofaec) had apparent relative molecular masses of 61,000 and 75,000, respectively, on sodium dodecyl sulfate-p ...200919242690
some studies of alpha-amylase production using aspergillus oryzae.the extracellular alpha-amylase production by aspergillus oryzae was studied in submerged fermentation using an adlof-kuhner orbital shaker. the effect of initial ph values in the range of 4 to 7.5 on enzyme production was investigated and initial ph medium of 6.2 +/- 0.1 resulted in enhanced alpha-amylase production. the effect of carbon and nitrogen source and composition was examined and it has been observed that corn starch concentration of 15 g l(-1) has sound effect on enzyme production. t ...200819260332
crystal structure of a fungal protease inhibitor from antheraea mylitta.indian tasar silk is produced by a wild insect called antheraea mylitta. insects do not have any antigen-antibody mediated immune system like vertebrates but they produce a wide variety of effector proteins and peptides possessing potent antifungal and antibacterial activity to combat microbial attack. antheraea mylitta expresses a fungal protease inhibitor amfpi-1, in the hemolymph that inhibits alkaline protease of aspergillus oryzae for protection against fungal infection. amfpi-1 is purified ...200919263521
crawler, a novel tc1/mariner-type transposable element in aspergillus oryzae transposes under stress conditions.a novel active transposable element, designated crawler, has been isolated from an industrial strain (osi1013) of aspergillus oryzae as an insertion sequence within the niad gene encoding nitrate reductase. it is 1290bp in length with imperfect terminal inverted repeats of 28bp and is flanked by 2bp (ta) target site duplications. it contains an open reading frame with no introns that encodes a putative transposase (aota) of 357 amino acid residues, which is highly homologous to the transposase e ...200919269345
morphological quantification of filamentous fungal development using membrane immobilization and automatic image analysis.mycelial morphology is a critically important process property in industrial fermentations of filamentous micro-organisms, as particular phenotypes are associated with maximum productivity. however, the accurate quantification of complex morphologies still represents a significant challenge in elucidating this relationship. a system has been developed for high-resolution characterisation of filamentous fungal growth on a solid substrate, using membrane immobilization and fully-automatic plug-ins ...200919277741
a novel transformation system using a bleomycin resistance marker with chemosensitizers for aspergillus oryzae.aspergillus oryzae is resistant to many kinds of antibiotics, which hampers their use to select transformants. in fact, the fungus is resistant to over 200microg/ml of bleomycin (bm). by enhancing the susceptibility of a. oryzae to bm using triton x-100 as a detergent and an atp-binding cassette (abc) pump inhibitor, chlorpromazine, to the growing medium, we established a novel transformation system by bm selection for a. oryzae. in a medium containing these reagents, a. oryzae showed little gro ...200919324021
analysis of microbial communities in doenjang, a korean fermented soybean paste, using nested pcr-denaturing gradient gel electrophoresis.doenjang is a traditional korean fermented soybean paste that provides a major source of protein. the microbial diversity of 10 samples of doenjang (5 commercially manufactured products and 5 homemade products) was investigated using nested pcr-denaturing gradient gel electrophoresis (dgge). in the first step, the nearly complete 16s rrna and 18s rrna genes were amplified using universal primers. subsequently, these products were used as a template in a nested pcr to obtain fragments suitable fo ...200919324443
deletion analysis of the promoter of aspergillus oryzae gene encoding heat shock protein order to find a promoter that could be influenced by temperature shift, we explored and isolated an aspergillus oryzae gene expressed at high temperatures (37-42 degrees c) by the cdna subtraction method. of the 96 cdna clones isolated from the subtraction library, one cdna clone showed 73% identity with aspergillus nidulans heat shock protein 30 (hsp30). based on this, we designated the isolated gene hsp30 of a. oryzae. a. oryzae hsp30 was weakly expressed at 30 degrees c, but strongly at 40 ...200919332290
development of biosensors containing laccase and imidazolium bis(trifluoromethylsulfonyl)imide ionic liquid for the determination of rutin.biosensors based on hydrophobic ionic liquids (ils) derived from the bis(trifluoromethylsulfonyl)imide [(cf(3)so(2))(2)n(-) = tf(2)n(-)] anion associated with three different imidazolium cations: 1-butyl-3-methylimidazolium (bmi x tf(2)n), 1-decyl-3-methylimidazolium (dmi x tf(2)n) and 1-tetradecyl-3-methylimidazolium (tdmi x tf(2)n), along with laccase from aspergillus oryzae, were constructed and optimized for determination of rutin. the laccase catalyzes the oxidation of rutin to the correspo ...200919345764
cloning and characterization of two flavohemoglobins from aspergillus oryzae.two flavohemoglobin (fhb) genes, fhb1 and fhb2, were cloned from aspergillus oryzae. the amino acid sequences of the deduced fhb1 and fhb2 showed high identity to other fhbs except for the predicted mitochondrial targeting signal in the n-terminus of fhb2. the recombinant proteins displayed absorption spectra similar to those of other fhbs. fhb1 and fhb2 were estimated to be a monomer and a dimer in solution, respectively. both of the isozymes exhibit high no dioxygenase (nod) activity. fhb1 uti ...200919351585
atan1p-an extracellular tannase from the dimorphic yeast arxula adeninivorans: molecular cloning of the atan1 gene and characterization of the recombinant enzyme.the tannase-encoding arxula adeninivorans gene atan1 was isolated from genomic dna by pcr, using as primers oligonucleotide sequences derived from peptides obtained after tryptic digestion of the purified tannase protein. the gene harbours an orf of 1764 bp, encoding a 587-amino acid protein, preceded by an n-terminal secretion sequence comprising 28 residues. the deduced amino acid sequence was similar to those of tannases from aspergillus oryzae (50% identity), a. niger (48%) and putative tann ...200919387973
evaluating the potential for enzymatic acrylamide mitigation in a range of food products using an asparaginase from aspergillus oryzae.asparaginase, an enzyme that hydrolyzes asparagine to aspartic acid, presents a potentially very effective means for reducing acrylamide formation in foods via removal of the precursor, asparagine, from the primary ingredients. an extracellular asparaginase amenable to industrial production was cloned and expressed in aspergillus oryzae . this asparaginase was tested in a range of food products, including semisweet biscuits, ginger biscuits, crisp bread, french fries, and sliced potato chips. in ...200919388639
a biotechnological product and its potential as a new immunomodulator for treatment of animal phlebovirus infection: punta toro virus.intracellular pathogens with widespread drug-resistance contribute substantially to the increasing rates in morbidity and mortality due to emerging and reemerging diseases. thus, the development of new drugs, including those that can enhance the immune response, is urgently needed. the immunomodulator, p-mapa, a proteinaceous aggregate of ammonium and magnesium phospholinoleate-palmitoleate anhydride derived from aspergillus oryzae, have been shown to induce antitumor activities. the ability of ...200919393266
genetic analysis of glucosidase ii beta-subunit in trimming of high-mannose-type glycans.glucosidase ii (g-ii) is a glycoprotein-processing enzyme that successively cleaves two alpha1,3-linked glucose residues from n-linked oligosaccharides in the endoplasmic reticulum. g-ii is a heterodimer whose alpha-subunit contains a glycosidase active site, but the function(s) of the beta-subunit remain poorly defined. we report here an in vivo enzymatic analysis using gene disruptants lacking either the g-ii alpha- or beta-subunit in the filamentous fungus aspergillus oryzae. using synthetic ...200919395677
heterologous expression and characterization of an endoglucanase from a symbiotic protist of the lower termite, reticulitermes speratus.rssymeg, an endoglucanase of glycosyl hydrolase family (ghf) 7 encoded by a transcript isolated from the symbiotic protist of the termite reticulitermes speratus, is expressed in aspergillus oryzae. interestingly, purified rssymeg1 has a relatively higher specific activity (603 micromol min(-1) mg(-1) protein) and v(max) value (769.6 unit/mg protein) than previously reported data for ghf7 endoglucanase of trichoderma ressei. it also has the same k(m) value (1.97 mg/ml) with clostridium celluloly ...201019404781
functional expression of the aspergillus flavus pks-nrps hybrid cpaa involved in the biosynthesis of cyclopiazonic acid.alpha-cyclopiazonic acid (cpa) is an indole tetramic acid mycotoxin. based on our identification of the polyketide synthase-nonribosomal peptide synthase (pks-nrps) hybrid gene cpaa involved in cyclopiazonic acid biosynthesis in aspergillus fungi, we carried out heterologous expression of aspergillus flavuscpaa under alpha-amylase promoter in aspergillus oryzae and identified its sole product to be the cpa biosynthetic intermediate cyclo-acetoacetyl-l-tryptophan (caatrp). this result rationalize ...200919410456
pepj is a new extracellular proteinase of aspergillus nidulans.under carbon starvation, aspergillus nidulans released a metallo-proteinase with activities comparable to those of prta, the major extracellular serine proteinase of the fungus. the relative molar mass of the enzyme was 19 kda as determined with both denaturing and renaturing sds page, while its isoelectric point and ph and temperature optima were 8.6, 5.5 and 65 degrees c, respectively. the enzyme was stable at ph 3.5-10.5 and was still active at 95 degrees c in the presence of azocasein substr ...200919418246
purification and identification of proteolytic enzymes from aspergillus oryzae capable of producing the antihypertensive peptide ile-pro-pro.two proteolytic enzymes capable of releasing the angiotensin i-converting enzyme (ace) inhibitor ile-pro-pro from casein were identified by purification of an aspergillus oryzae extract by three-step column chromatography. first, proteins capable of producing ile-pro-pro from beta-casein were eluted using a deae-sepharose ff column with a linear sodium chloride gradient. an endopeptidase capable of releasing pro-ile-pro-gln-ser-leu-pro-gln-asn-ile-pro-pro from pro-ile-pro-gln-ser-leu-pro-gln-asn ...200919447337
an acidophilic beta-galactosidase from bispora sp. mey-1 with high lactose hydrolytic activity under simulated gastric conditions.bgala, a full-length gene (3,009 bp) that encodes a beta-galactosidase, was cloned from the meso-acidophilic fungus bispora sp. mey-1 and expressed in pichia pastoris. the deduced amino acid sequence of bgala shares highest identity (55.5%) with the beta-galactosidase from aspergillus phoenicis, which belongs to the glycoside hydrolyase family 35. purified recombinant bgala is acidophilic, exhibiting maximum activity at ph 1.5, which is lower than that reported for other beta-galactosidases. the ...200919453169
comparison of stirred tank and airlift bioreactors in the production of polygalacturonases by aspergillus oryzae.the production of endo and exo-polygalacturonase (pg) by aspergillus oryzae ipt 301 was studied in a stirred tank bioreactor (str) and an internal circulation airlift bioreactor. using a factorial experimental design, a soluble culture medium was defined which allowed the production of exo- and endo-pg comparable to that obtained in a medium containing suspended wheat bran. the soluble medium was used in tests to compare the production of these enzymes in the str and airlift bioreactor. in these ...200919467860
crystallization of carbohydrate oxidase from microdochium nivale.microdochium nivale carbohydrate oxidase was produced by heterologous recombinant expression in aspergillus oryzae, purified and crystallized. the enzyme crystallizes with varying crystal morphologies depending on the crystallization conditions. several different crystal forms were obtained using the hanging-drop vapour-diffusion method, two of which were used for diffraction measurements. hexagon-shaped crystals (form i) diffracted to 2.66 a resolution, with unit-cell parameters a = b = 55.7, c ...200919478452
alpha-lactalbumin hydrolysate stimulates glucagon-like peptide-2 secretion and small intestinal growth in suckling rats.we investigated whether bovine milk constituents influenced glucagon-like peptide (glp)-2 secretion and intestinal growth in suckling rats. male sprague-dawley rats (14 d old) received i.g. infusions of a milk protein fraction, a lactose solution, or the cream fraction of milk. the serum concentration of glp-2, but not glp-1, markedly increased in rats administered milk protein compared with those given the lactose solution or the cream fraction from 60 to 120 min after administration. in anothe ...200919494023
a unique post-translational processing of an exo-beta-1,3-glucanase of penicillium sp. kh10 expressed in aspergillus characterize an exo-beta-1,3-glucanase (exgp) of an isolated fungal strain with high laminarin degradation activity, identified as penicillium sp. kh10, heterologous secretory expression of the exgp was performed in aspergillus oryzae. deduced amino acid sequence of the exgp gene possibly consisted of 989 amino acids which showed high sequence similarity to those of fungal exo-beta-1,3-glucanases belonging to the glycoside hydrolase (gh) family 55. notably, the purified recombinant exgp showe ...200919500674
is electron equivalence between substrate and product preferable to c-mol equivalence in representations of microbial anabolism applicable to "origin of life" environmental conditions.this research compares two techniques for constructing sets of equations representing the growth of a microorganism on a given substrate. each anabolic equation represents a defined initial and a final state. the initial state comprises all necessary mineral nutrients and one organic source of carbon and energy. the final state comprises the microbial cells that have grown, and any additional organic or inorganic products of the growth process. the cells are represented by a unit-carbon formula ...200919501106
functional analysis and subcellular localization of two geranylgeranyl diphosphate synthases from penicillium paxilli.the filamentous fungus penicillium paxilli contains two distinct geranylgeranyl diphosphate (ggpp) synthases, ggsa and ggsb (paxg). paxg and its homologues in neotyphodium lolii and fusarium fujikuroi are associated with diterpene secondary metabolite gene clusters. the genomes of other filamentous fungi including aspergillus fumigatus, aspergillus nidulans, aspergillus niger, aspergillus oryzae and fusarium graminearum also contain two or more copies of ggpp synthase genes, although the diterpe ...200919529962
cereal-based biorefinery development: utilisation of wheat milling by-products for the production of succinic acid.a novel wheat-based bioprocess for the production of a nutrient-complete feedstock for the fermentative succinic acid production by actinobacillus succinogenes has been developed. wheat was fractionated into bran, middlings and flour. the bran fraction, which would normally be a waste product of the wheat milling industry, was used as the sole medium in two solid-state fermentations (ssf) of aspergillus awamori and aspergillus oryzae that produce enzyme complexes rich in amylolytic and proteolyt ...200919539669
modified electrophoretic and digestion conditions allow a simplified mass spectrometric evaluation of disulfide bonds.proper formation of disulfide bonds in proteins is a prerequisite to their stability and function. information on disulfide pattern may therefore serve as an indication of the proper folding of recombinant proteins, and can also be used in protein homology modeling for the purpose of structure refinement. protein handling and digestion at basic ph leads to disulfide bond scrambling. that is why the samples are usually treated and digested at low ph where no scrambling occurs. unfortunately, the ...200919551705
heterologous expression and characterization of cpi, ocpa, and novel serine-type carboxypeptidase ocpb from aspergillus the genome of aspergillus oryzae, 12 genes have been predicted to encode serine-type carboxypeptidases. however, the carboxypeptidase activities of the proteins encoded by these genes have not yet been confirmed experimentally. in this study, we have constructed three of these 12 genes overexpressing strains using aspergillus nidulans and characterized their overproduced recombinant proteins. of these three genes, one was previously named cpi; the other two have not been reported yet, and hen ...200919557408
responses of aspergillus oryzae to stimuli from near-uv light irradiation in the presence or absence of titanium dioxide.the responses of aspergillus oryzae to the stimuli from near-uv irradiation were investigated by using a black light fluorescent lamp in the presence or absence of tio2 particles. light irradiation at an intensity of 6 w/m2 strongly inhibited the growth of germinated pellets of a. oryzae. this growth inhibition was weakened by tio2 particles (0.05 g/l), especially in the initial growth phase, in which the expression level of catalase gene (catb) was approximately three times higher than that in ...200919579656
[promotive excretion of causative agents of yusho by intake of fermented brown rice with aspergillus oryze in patients with yusho --with regard to pcdfs and pcdds].forty years have passed since the outbreak of kanemi rice oil poisoning, namely, yusho in the western japan. however, even now the patients with yusho have been still suffering from several objective and subjective symptoms. in order to improve or, if possible, to cure such symptoms, the most important therapeutic treatment is considered to actively excrete the causative agents, that is, polychlorinated dibenzofurans (pcdfs) and polychlorinated dibenzo-p-dioxins (pcdds) from the bodies of the pa ...200919588849
d-lactic acid production from cellooligosaccharides and beta-glucan using l-ldh gene-deficient and endoglucanase-secreting lactobacillus order to achieve direct fermentation of an optically pure d: -lactic acid from cellulosic materials, an endoglucanase from a clostridium thermocellum (cela)-secreting plasmid was introduced into an l: -lactate dehydrogenase gene (ldhl1)-deficient lactobacillus plantarum (ldhl1) bacterial strain. cela expression and its degradation of beta-glucan was confirmed by western blot analysis and enzyme assay, respectively. although the cela-secreting ldhl1 assimilated cellooligosaccharides up to cell ...201019597813
post-genomic insights into the plant polysaccharide degradation potential of aspergillus nidulans and comparison to aspergillus niger and aspergillus oryzae.the plant polysaccharide degradative potential of aspergillus nidulans was analysed in detail and compared to that of aspergillus niger and aspergillus oryzae using a combination of bioinformatics, physiology and transcriptomics. manual verification indicated that 28.4% of the a. nidulans orfs analysed in this study do not contain a secretion signal, of which 40% may be secreted through a non-classical method.while significant differences were found between the species in the numbers of orfs ass ...200919618505
release of antihypertensive peptides in miso paste during its fermentation, by the addition of casein.casein hydrolysate by aspergillus oryzae protease has been developed to obtain two kinds of antihypertensive peptides, val-pro-pro and ile-pro-pro. in this study, casein miso paste was prepared by adding casein at various concentrations during miso paste fermentation in order to release angiotensin-i converting enzyme (ace) inhibitory peptides. increase of peptide concentration and ace inhibitory activity in casein miso paste was observed and corresponded to the degradation of casein and soy bea ...200919619856
cloning of aspergillus oryzae aovps5 gene, homologous to vacuolar protein sorting associated gene vps5 and construction of the disruptant.aovps5 gene was isolated from aspergillus oryzae as a homologue to s. cerevisiae vps5 gene which encodes a polypeptide consisting of 451 amino acids that is nearly 32% homologous to vps5p. three aovps5 gene disruptants were generated and they showed higher activity of tripeptidyl peptidase, which is mainly detected in vacuoles, in their culture medium. higher amount of nitrogenous constituent was found in the filtrate of wheat gluten degraded by addition of culture medium of these disruptants th ...200919619858
a food-grade enzyme preparation with modest gluten detoxification properties.celiac sprue is a life-long disease characterized by an intestinal inflammatory response to dietary gluten. a gluten-free diet is an effective treatment for most patients, but accidental ingestion of gluten is common, leading to incomplete recovery or relapse. food-grade proteases capable of detoxifying moderate quantities of dietary gluten could mitigate this problem.200919621078
identification of a basic helix-loop-helix-type transcription regulator gene in aspergillus oryzae by systematically deleting large chromosomal segments.we previously developed two methods (loop-out and replacement-type recombination) for generating large-scale chromosomal deletions that can be applied to more effective chromosomal engineering in aspergillus oryzae. in this study, the replacement-type method is used to systematically delete large chromosomal dna segments to identify essential and nonessential regions in chromosome 7 (2.93 mb), which is the smallest a. oryzae chromosome and contains a large number of nonsyntenic blocks. we constr ...200919633118
characterization of a neutral ceramidase orthologue from aspergillus oryzae.ceramide is an important molecule not only structurally but also regulationally as a modulator of various cellular events. ceramidase (cdase) are classified into three different types (acid, alkaline, and neutral cdases). neutral cdase could play an important role in the regulation of ceramide levels in the extracellular space. in this study, we describe the characterization of a neutral cdase orthologue from the filamentous fungus aspergillus oryzae. the gene encoding the neutral cdase ortholog ...200919650849
characterization of a 52 kda exoantigen of penicillium chrysogenum and monoclonal antibodies suitable for its detection.the indoor clade of penicillium chrysogenum, the so-called fleming clade, is the most common species of penicillium on moldy building materials. in a previous study, we identified a 52 kda human antigen characteristic of the indoor clade of p. chrysogenum not present in a taxonomically diverse selection of fungi. further investigations revealed that it is a modestly glycosylated mature protein with a pi 5.3. the protein is apparently identical to a glucoamylase previously reported from an alumin ...201019653120
nonhomologous end-joining deficiency allows large chromosomal deletions to be produced by replacement-type recombination in aspergillus oryzae.gene targeting is a technique of introducing a genetic trait at a predetermined site within a genome; it is also used to eliminate undesirable chromosomal regions from the relevant genome. thus far, replacement-type recombination between two homologous regions separated by a large nonhomologous sequence has been hardly achieved probably due to the low frequency of homologous recombination in filamentous fungi. in this study, we report the successful and highly efficient deletion by replacement-t ...200919654050
isolation of a new metabolite from biotransformation of daidzein by aspergillus oryzae.two compounds were isolated from biotransformation of daidzein, a soybean isoflavone, by aspergillus oryzae. one was 6,7,8,4'-tetrahydroxyisoflavone, an a-ring dihydroxylated daidzein, and the other was a novel compound which had the cleaved b-ring of daidzein. the former was perhaps derived from 8-hydroxydaidzein and/or 6-hydroxydaidzein, and had high dpph radical-scavenging activity.200919661709
genome-wide analysis of maltose utilization and regulation in aspergilli.maltose utilization and regulation in aspergilli is of great importance for cellular physiology and industrial fermentation processes. in aspergillus oryzae, maltose utilization requires a functional mal locus, composed of three genes: malr encoding a regulatory protein, malt encoding maltose permease and mals encoding maltase. through a comparative genome and transcriptome analysis we show that the mal regulon system is active in a. oryzae while it is not present in aspergillus niger. in order ...200919696104
endocytic recycling at the tip region in the filamentous fungus aspergillus oryzae.recent live cell imaging analyzing the components required for endocytosis has elucidated that endocytosis actively occurs at the hyphal tip region in filamentous fungi. to examine further the physiological roles of endocytosis we investigated a conditional mutant of endocytosis in aspergillus oryzae. endocytosis-deficient hyphae displayed retarded apical growth, abnormal hyphal morphology, mislocalization of a vesicle- snare, which is thought to undergo endocytic recycling to the tip region, an ...200919721880
production and characterization of a milk-clotting enzyme from aspergillus oryzae mtcc 5341.microbial milk-clotting enzymes are valued as calf rennet substitutes in the cheese industry. aspergillus oryzae mtcc 5341 was identified to produce the highest milk-clotting activity during screening of 16 fungal strains. solid state fermentation using wheat bran along with 4% defatted soy flour and 2% skim milk powder as substrate was optimal for growth of a. oryzae and production of the enzyme. nearly 40,000 u/g bran of milk-clotting activity was present at the end of 120 h. the enzyme could ...201019727708
characterization and kinetic analysis of a thermostable gh3 beta-glucosidase from penicillium brasilianum.a gh3 beta-glucosidase (bgl) from penicillium brasilianum was purified to homogeneity after cultivation on a cellulose and xylan rich medium. the bgl was identified in a genomic library, and it was successfully expressed in aspergillus oryzae. the bgl had excellent stability at elevated temperatures with no loss in activity after 24 h of incubation at 60 degrees c at ph 4-6, and the bgl was shown to have significantly higher stability at these conditions in comparison to novozym 188 and to other ...201019756584
aspergillus oryzae atfa controls conidial germination and stress tolerance.we compared atfa and atfb, the genes encoding the respective atf/creb-type transcription factors in aspergillus oryzae. the germination ratio of deltaatfa conidia was low without any stress, unlike that of deltaatfb conidia. the deltaatfa conidia were more sensitive to oxidative stress than the deltaatfb conidia, which are also sensitive to oxidative stress. we compared the gene expressions of these strains by using a dna microarray, genechip. almost all the genes regulated by atfb were also reg ...200919770065
genes regulated by aoxlnr, the xylanolytic and cellulolytic transcriptional regulator, in aspergillus oryzae.xlnr is a zn(ii)2cys6 transcriptional activator of xylanolytic and cellulolytic genes in aspergillus. overexpression of the aoxlnr gene in aspergillus oryzae (a. oryzae xlnr gene) resulted in elevated xylanolytic and cellulolytic activities in the culture supernatant, in which nearly 40 secreted proteins were detected by two-dimensional electrophoresis. dna microarray analysis to identify the transcriptional targets of aoxlnr led to the identification of 75 genes that showed more than fivefold i ...200919777228
uncovering transcriptional regulation of glycerol metabolism in aspergilli through genome-wide gene expression data analysis.glycerol is catabolized by a wide range of microorganisms including aspergillus species. to identify the transcriptional regulation of glycerol metabolism in aspergillus, we analyzed data from triplicate batch fermentations of three different aspergilli (aspergillus nidulans, aspergillus oryzae and aspergillus niger) with glucose and glycerol as carbon sources. protein comparisons and cross-analysis with gene expression data of all three species resulted in the identification of 88 genes having ...200919784673
discovery of a new tyrosinase-like enzyme family lacking a c-terminally processed domain: production and characterization of an aspergillus oryzae catechol oxidase.a homology search against public fungal genome sequences was performed to discover novel secreted tyrosinases. the analyzed proteins could be divided in two groups with different lengths (350-400 and 400-600 residues), suggesting the presence of a new class of secreted enzymes lacking the c-terminal domain. among them, a sequence from aspergillus oryzae (408 aa, aoco4) was selected for production and characterization. aoco4 was expressed in trichoderma reesei under the strong cbh1 promoter. expr ...201019798497
identification of two aflatrem biosynthesis gene loci in aspergillus flavus and metabolic engineering of penicillium paxilli to elucidate their function.aflatrem is a potent tremorgenic toxin produced by the soil fungus aspergillus flavus, and a member of a structurally diverse group of fungal secondary metabolites known as indole-diterpenes. gene clusters for indole-diterpene biosynthesis have recently been described in several species of filamentous fungi. a search of aspergillus complete genome sequence data identified putative aflatrem gene clusters in the genomes of a. flavus and aspergillus oryzae. in both species the genes for aflatrem bi ...200919801473
acrylamide degradation by filamentous fungi used in food and beverage industries.filamentous fungi (24 strains) used in food and beverage industries were investigated for acrylamide-degradation ability: aspergillus oryzae kbn1010 showed the highest ability. little acrylic acid was produced but no glycidamide was detected during aa degradation in roasted green tea; therefore, a. oryzae could be used for reducing the aa concentration.200919804862
analysis of enzyme production by submerged culture of aspergillus oryzae using whole barley.we have reported on high enzyme production by submerged culture of aspergillus kawachii using barley with the husk (whole barley). to elucidate the mechanism underlying this high enzyme production, we performed a detailed analysis. aspergillus oryzae rib40 was submerged-cultured using whole barley and milled whole barley. enzyme production was analyzed in terms of changes in medium components and gene expression levels. when whole barley was used, high production of glucoamylase and alpha-amylas ...200919809198
structural and functional studies of aspergillus oryzae cutinase: enhanced thermostability and hydrolytic activity of synthetic ester and polyester degradation.cutinases are responsible for hydrolysis of the protective cutin lipid polyester matrix in plants and thus have been exploited for hydrolysis of small molecule esters and polyesters. here we explore the reactivity, stability, and structure of aspergillus oryzae cutinase and compare it to the well-studied enzyme from fusarium solani. two critical differences are highlighted in the crystallographic analysis of the a. oryzae structure: (i) an additional disulfide bond and (ii) a topologically favor ...200919810726
biosensor for luteolin based on silver or gold nanoparticles in ionic liquid and laccase immobilized in chitosan modified with cyanuric chloride.novel and effective biosensors based on ag or au nanoparticles dispersed in ionic liquid (il) 1-butyl-3-methylimidazolium hexafluorophosphate ( and laccase (lac) from aspergillus oryzae immobilized in chitosan (chi) chemically cross-linked with cyanuric chloride (cc) were constructed. this enzyme catalyzes the oxidation of luteolin to the corresponding o-quinone, which is electrochemically reduced back to luteolin at 0.35 v vs. ag/agcl. square-wave voltammetry was used for the electroc ...200919838422
genetic analysis of conidiation regulatory pathways in koji-mold aspergillus oryzae.conidia of koji-mold aspergillus oryzae are often used as starters in the fermented food industry. however, little is known about conidiation regulation in a. oryzae. to improve the productivity of conidia in a. oryzae, it is necessary to understand conidiation regulation in the strain. therefore, we analyzed the conidiation regulatory system in a. oryzae using 10 kinds of conidiation regulatory gene disruptants. the phenotypes of aorflug, aorflba, aorflbb, aorflbc, aorflbd, aorflbe, aorbrla, ao ...201019850144
characterization and expression analysis of a maltose-utilizing (mal) cluster in aspergillus oryzae.starch and maltooligosaccharides such as maltose and maltotriose induce the production of amylolytic enzymes including alpha-amylase in aspergillus oryzae. a transcriptional activator gene amyr, required for maltose induction of amylolytic enzymes, has been cloned and characterized. the amyr gene deletion mutant showed significantly poor growth on starch medium but normal growth on maltose medium. this indicated the existence of another maltose-utilizing system, whose expression might not be con ...201019850146
a trial of minimization of chromosome 7 in aspergillus oryzae by multiple chromosomal deletions.we aim to create an aspergillus oryzae mutant with a highly reduced chromosome, but better growth, by eliminating the nonessential regions coding various dispensable functions for its better industrial use. in our previous study, we successfully determined the outline of essential and nonessential regions by constructing a series of large chromosomal deletions in a. oryzae chromosome 7. based on these results, we here constructed two mutants, designated rkuaf7a and rkuaf7b, lacking 24.7 and 24% ...201019855999
characterization of cyclo-acetoacetyl-l-tryptophan dimethylallyltransferase in cyclopiazonic acid biosynthesis: substrate promiscuity and site directed mutagenesis studies.the fungal neurotoxin alpha-cyclopiazonic acid (cpa), a nanomolar inhibitor of ca(2+)-atpase with a unique pentacyclic indole tetramic acid scaffold, is assembled by a three enzyme pathway cpas, cpad, and cpao in aspergillus sp. we recently characterized the first pathway-specific enzyme cpas, a hybrid two module polyketide synthase-nonribosomal peptide synthetase (pks-nrps) that generates cyclo-acetoacetyl-l-tryptophan (caatrp). here we report the characterization of the second pathway-specific ...200919877600
a cytosolic phospholipase a2-like protein in the filamentous fungus aspergillus oryzae localizes to the intramembrane space of the mammalian cells, cytosolic phospholipase a(2) (cpla(2)) displays a variety of activities through the release of arachidonic acid in response to cellular stimuli. in this study, we characterize the putative cpla(2)-like protein, aoplaa, in the filamentous fungus aspergillus oryzae. when aoplaa-egfp was expressed in a. oryzae, it localized to the tubular structures that was costained by the marker dye for the mitochondria. a biochemical fractionation experiment showed that aoplaa was present in ...200919889028
characterization of galactosyl derivatives obtained by transgalactosylation of lactose and different polyols using immobilized beta-galactosidase from aspergillus oryzae.the synthesis of novel galactosides is interesting because of their important role in several biological processes. their properties greatly depend upon the configuration and type of galactoside. therefore, to study biological activity, it is essential to elucidate the structure of the products. glycosidases are capable of catalyzing glycosidic linkages with absolute stereoselectivity of the anomeric center. we report the enzymatic synthesis of galactosyl-ethylene glycol, galactosyl-glycerol, an ...200919891502
improvement of heterologous protein production in aspergillus oryzae by rna interference with alpha-amylase genes.aspergillus oryzae rib40 has three alpha-amylase genes (amya, amyb, and amyc), and secretes alpha-amylase abundantly. however, large amounts of endogenous secretory proteins such as alpha-amylase can compete with heterologous protein in the secretory pathway and decrease its production yields. in this study, we examined the effects of suppression of alpha-amylase on heterologous protein production in a. oryzae, using the bovine chymosin (chy) as a reporter heterologous protein. the three alpha-a ...200919897917
analysis of bacterial and fungal communities in japanese- and chinese-fermented soybean pastes using nested pcr-dgge.the microbial diversity of japanese- and chinese-fermented soybean pastes was investigated using nested pcr-denaturing gradient gel electrophoresis (dgge). five japanese-fermented soybean paste samples and three chinese-fermented soybean paste samples were analyzed for bacteria and fungi. extracted dna was used as a template for pcr to amplify 16s rrna and 18s rrna genes. the nearly complete 16s rrna and 18s rrna genes were amplified using universal primers, and the resulting products were subse ...201019924476
acid protease production by solid-state fermentation using aspergillus oryzae mtcc 5341: optimization of process parameters.aspergillus oryzae mtcc 5341, when grown on wheat bran as substrate, produces several extracellular acid proteases. production of the major acid protease (constituting 34% of the total) by solid-state fermentation is optimized. optimum operating conditions obtained are determined as ph 5, temperature of incubation of 30 degrees c, defatted soy flour addition of 4%, and fermentation time of 120 h, resulting in acid protease production of 8.64 x 10(5) u/g bran. response-surface methodology is used ...201019937364
using a new gpi-anchored-protein identification system to mine the protein databases of aspergillus fumigatus, aspergillus nidulans, and aspergillus oryzae.computational approaches provide valuable information to start experimental surveys identifying glycosylphosphatidylinositol (gpi)-anchored proteins in protein sequence databases. we developed a new sequence-based identification system that uses an optimized classifier based on a support vector machine (svm) algorithm to recognize appropriate cooh-terminal sequences and uses a classifier implementing a simple majority voting strategy to recognize appropriate nh2-terminal sequences. the svm class ...200919940384
aoso protein accumulates at the septal pore in response to various stresses in the filamentous fungus aspergillus oryzae.filamentous ascomycetes form hyphal networks that are compartmentalized by septa which have a perforated pore allowing the passage of cytoplasm and organelles between adjacent hyphal compartments. thus, the septal pore may play an important role in the organized growth of multicellular organisms. upon hyphal injury, the septal pore is plugged by a wound-healing organelle, known as the woronin body, to prevent excessive cytoplasmic leakage. however, the movement of proteins towards the septal por ...201019945422
hydrolysis of amylopectin by amylolytic enzymes: structural analysis of the residual amylopectin population.amylopectin fine structures were studied following limited hydrolysis of gelatinised waxy maize starch by amylases with a different level of inner chain attack (lica). this was done by size exclusion chromatography as well as by debranching the (partially hydrolysed) amylopectin samples and studying the size distributions of the released chains. alpha-amylases from bacillus amyloliquefaciens and aspergillus oryzae, with a relatively high lica, drastically altered amylopectin chain length distrib ...201019962130
hydrolysis of amylopectin by amylolytic enzymes: level of inner chain attack as an important analytical differentiation criterion.differences in amylase action pattern on amylopectin were demonstrated by the relation between the decrease in potassium iodide-iodine binding of waxy maize starch and the increase in reducing value during hydrolysis, as expressed by the rv(80) value (i.e., the reducing value for a potassium iodide-iodine binding value of 80% of that of the starting material). in the initial stages of the hydrolysis, the ratio of the increase in the level of reducing polysaccharides to the increase in the total ...201019962134
expression in escherichia coli of an unnamed protein gene from aspergillus oryzae rib40 and cofactor analyses of the gene product as formate unnamed protein of aspergillus oryzae rib40 (accession no. xp_001727378), the amino acid sequence of which shows high similarity to those of formate oxidase isoforms produced by debaryomyces vanjiriae mh201, was produced in escherichia coli in c-his(6)-tagged form. the gene product, purified by affinity column chromatography, catalyzed the oxidation of formate to yield hydrogen peroxide but showed no evidence of activity on the other substrates tested. the k(m) and v(max) values at 30 degrees ...200919966484
simultaneous preparation of purified plasmalogens and sphingomyelin in human erythrocytes with phospholipase a1 from aspergillus orizae.a method for the simultaneous purification of plasmalogens and sphingomyelin (sm) in human erythrocytes is described. treatment of total lipids with n-hexane/acetone (1:1 v/v) resulted in selective precipitation of sm. both the supernatant and the precipitate fractions were incubated with a phospholipase a(1) (pla1) from aspergillus orizae for 3.5 h. the pla1-treated lipids were extracted with n-hexane/isopropanol, the hexane layer was obtained using a na(2)so(4) solution, and the hexane layer w ...200919966491
enzymatic degradation products from a marine polysaccharide ycp with different immunological activity and binding affinity to macrophages, hydrolyzed by alpha-amylases from different origins.ycp is a marine polysaccharide with anti-tumor and immune-modulating effects. this study evaluated the effect of enzymatic degradation of ycp by alpha-amylases from different origins on its immunological activity and binding ability to the macrophages. ycp was hydrolyzed by alpha-amylases isolated from aspergillus oryzae, bacillus licheniformis, barley malt, and porcine pancreas respectively, then four fragments with unique molecular weight (termed: ycp-ao, ycp-bl, ycp-bm, and ycp-pp, respective ...201020004229
production of fructooligosaccharides by beta-fructofuranosidases from aspergillus oryzae kb.aspergillus oryzae kb produces two types of beta-fructofuranosidases: f1 and f2. f1 produces the fructooligosaccharides (foss) 1-kestose, nystose, and fructosyl nystose from sucrose through a transfructosylation action, whereas f2 mainly hydrolyzes sucrose to glucose and fructose. f1 and f2 enzymes were more selectively produced from the kb strain in liquid media with a sucrose concentration>2% and <2%, respectively. immobilization using an anion-exchange resin (wa-30; polystyrene with tertiary ...201020014851
characterization of recombinant prolyl aminopeptidase from aspergillus oryzae.prolyl aminopeptidase (pap) degrades only amino-terminal proline from peptides. the food-grade fungus aspergillus oryzae produces this enzyme only in small amounts. in this paper, we present efficient production of recombinant pap with an overexpression system of a. oryzae and characterization of its biochemical properties.201020028436
distinct enzymatic and cellular characteristics of two secretory phospholipases a2 in the filamentous fungus aspergillus oryzae.microbial secretory phospholipases a(2) (spla(2)s) are among the last discovered and least known members of this functionally diverse family of enzymes. we analyzed here two spla(2)s, named splaa and splab, of the filamentous ascomycete aspergillus oryzae. splaa and splab consist of 222 and 160 amino acids, respectively, and share the conserved cys and catalytic his-asp residues typical of microbial spla(2)s. two spla(2)s differ in ph optimum, ca(2+) requirement and expression profile. the splaa ...201020045482
bioreduction of trivalent aurum to nano-crystalline gold particles by active and inactive cells and cell-free extract of aspergillus oryzae var. viridis.bioreduction efficacy of both active (ab) and inactive (ib) cells/biomass of aspergillus oryzae var. viridis and their respective cell-free extracts (ace and ice) to convert trivalent aurum to gold nanoparticles were tested in the present study. strong plasmon resonance of gold nanoparticles was observed between 540 and 560 nm in the samples obtained from ab, ib, ace and ice. transmission electron microscopy (tem), field emission scanning electron microscopy (fe-sem), energy dispersive x-ray (ed ...201020056324
characterization of the palh gene from aspergillus oryzae.we disrupted the palh gene, which is known to participate in the ambient ph signal transduction pathway, in aspergillus oryzae. palh disruption caused significant decreases in pacc expression and alkaline protease activity. hence we believe that palh plays a very important role in controlling the alkaline protease level in a. oryzae.201020057127
evaluation of enzyme mixtures in releasing fermentable sugars from pre-pulping extracts of mixed northeast near-term option to developing a forest product biorefinery is to derive pre-pulping extract from incoming wood chips before the main pulping step. the release of monomer sugars from a xylan-rich extract, creating a fermentable substrate is a prerequisite for utilization of pre-pulping extract for production of ethanol or other value-added products. this study examined the individual and mixture efficiencies of two hemicellulolytic microbial enzymes and two xylanase preparations in catalyzin ...201020084471
characterization of aroma compounds in chinese rice wine qu by solvent-assisted flavor evaporation and headspace solid-phase microextraction.the aroma of chinese rice wine qu is one of the most important factors that influences the flavor of chinese rice wine. to better understand the aroma of qu, aroma compounds in four wheat qus and two xiao qus were identified by chromatography-olfactometry (gc-o) after solvent extraction followed by solvent-assisted flavor evaporation (safe). a total of 39 aroma compounds were characterized by gc-o. on the basis of aroma intensity, 1-hexanal, ethyl hexanoate, 1-octen-3-ol, and phenylacetaldehyde ...201020088505
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