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discovery of a linoleate 9s-dioxygenase and an allene oxide synthase in a fusion protein of fusarium oxysporum.fusarium oxysporum is a devastating plant pathogen that oxidizes c₁₈ fatty acids sequentially to jasmonates. the genome codes for putative dioxygenase (dox)-cytochrome p450 (cyp) fusion proteins homologous to linoleate diol synthases (ldss) and the allene oxide synthase (aos) of aspergillus terreus, e.g., foxb_01332. recombinant foxb_01332 oxidized 18:2n-6 to 9s-hydroperoxy-10(e),12(z)-octadecadienoic acid by hydrogen abstraction and antarafacial insertion of molecular oxygen and sequentially to ...201324082064
fatal-mixed cutaneous zygomycosis-aspergillosis: a case report.mucorales and aspergillus are molds responsible for infections in immunocompromised patients. in this report, we describe a case of a rare extensively mixed cutaneous infection caused by lichtheimia ramosa, aspergillus fumigatus and aspergillus terreus in a neutropenic patient suffering from an acute leukemia. the fatal outcome of this patient can be attributed to its hematologic malignancy, the extensive nature of the lesions and the resistance of the strains to antifungals.201324085614
rapid determination of lovastatin in the fermentation broth of aspergillus terreus using dual-wavelength uv spectrophotometry.lovastatin, a hypocholesterolemic drug, is produced by submerged fermentation of aspergillus terreus thom (trichocomaceae). high performance liquid chromatography is usually used to determine lovastatin in samples of the fermentation broth. however, this method is inconvenient and costly, especially in the context of high-throughput sample analysis.201424093797
[aspergillus species in hospital environments with pediatric patients in critical condition].aspergillus is a group of opportunistic fungi that cause infections, with high morbimortality in immunosuppressed patients. aspergillus fumigatus is the most frequent species in these infections, although the incidence of other species has increased in the last few years.201424120420
2-hydroxyisocaproic acid is fungicidal for candida and aspergillus species.the amino acid derivative 2-hydroxyisocaproic acid (hica) is a nutritional additive used to increase muscle mass. low levels can be detected in human plasma as a result of leucine metabolism. it has broad antibacterial activity but its efficacy against pathogenic fungi is not known. the aim was to test the efficacy of hica against candida and aspergillus species. efficacy of hica against 19 clinical and reference isolates representing five candida and three aspergillus species with variable azol ...201424125484
biodegradation of a mixture of pahs by non-ligninolytic fungal strains isolated from crude oil-contaminated soil.nine native non-ligninolytic fungal strains were isolated from maya crude oil-contaminated soil and selected based on their ability to grow and use crude oil and several polycyclic aromatic hydrocarbons (pahs) as carbon source, for their application to pah removal in soil. the fungi were identified by pcr amplification of intergenic transcribed sequences regions and microbiological techniques, and results showed them to be part of the genera fusarium, neurospora, aspergillus, scedosporium, penic ...201424132496
unusual biotransformation products of the sesquiterpene lactone budlein a by aspergillus species.biotransformation of chemicals by microorganisms can be effective in increasing chemical diversity. some fungi have been described to be useful for the biotransformation of sesquiterpene lactones. nevertheless, in most cases, only minor or simple transformations of functional groups have been observed. budlein a is a sesquiterpene lactone found in high amounts in american sunflower-like species of the genus viguiera (asteraceae). it shows important biological effects like in vitro and in vivo an ...201324135634
relationship between morphology and itaconic acid production by aspergillus terreus.the morphology of filamentous fungi closely correlates with the productivity in submerged culture. using itaconic acid (ia) production by aspergillus terreus as a research model, the quantitative relationship between the growth form of a. terreus and ia production was investigated. ia fermentation was scaled up from shake flasks to a 7 l stirred tank bioreactor based on the quantitative relationship. our results demonstrated the following: (1) three morphologies of a. terreus were formed by chan ...201424169454
identification of ochratoxin a producing fungi associated with fresh and dry liquorice.the presence of fungi on liquorice could contaminate the crop and result in elevated levels of mycotoxin. in this study, the mycobiota associated with fresh and dry liquorice was investigated in 3 producing regions of china. potential toxigenic fungi were tested for ochratoxin a (ota) and aflatoxin b1 (afb1) production using liquid chromatography/mass spectrometry/mass spectrometry. based on a polyphasic approach using morphological characters, β-tubulin and rna polymerase ii second largest subu ...201324205182
astpt catalyses both reverse n1- and regular c2 prenylation of a methylated bisindolyl benzoquinone.prenylated bisindolyl benzoquinones exhibit interesting biological activities, such as antidiabetic or anti-hiv activities. a number of these compounds, including asterriquinones, have been isolated from aspergillus terreus. in this study, we identified two putative genes by genome mining, ateg_09980 and ateg_00702, which share high sequence similarity with the known bisindolyl benzoquinone prenyltransferase tdib from aspergillus nidulans. the coding sequences were cloned and overexpressed in e. ...201424302698
a systems biology approach for the identification of target genes for the improvement of itaconic acid production in aspergillus species.in this paper, a clone based transcriptome analysis towards the identification of genes related to itaconic acid production in aspergillus terreus was carried out as an extension of a previously published a clone-based transcriptome analysis from a set of batch fermentation experiments. also a publically available a. niger transcriptome dataset from cultures similar to those of the a. terreus data set was analyzed to evaluate the specificity of the approach followed for a. terreus.201324304666
cloning, characterization and application of a glyceraldehyde-3-phosphate dehydrogenase promoter from aspergillus terreus.it is important to develop native and highly efficient promoters for effective genetic engineering of filamentous fungi. although aspergillus terreus is an important industrial fungus for the production of itaconic acid and lovastatin, the available genetic toolbox for this microorganism is still rather limited. we have cloned the 5' upstream region of the glyceraldehyde-3-phosphate dehydrogenase gene (gpd; 2,150 bp from the start codon) from a. terreus cicc 40205 and subsequently confirmed its ...201424306453
characterization of the co-purified invertase and β-glucosidase of a multifunctional extract from aspergillus terreus.the filamentous fungus aspergillus terreus secretes both invertase and β-glucosidase when grown under submerged fermentation containing rye flour as the carbon source. the aim of this study was to characterize the co-purified fraction, especially the invertase activity. an invertase and a β-glucosidase were co-purified by two chromatographic steps, and the isolated enzymatic fraction was 139-fold enriched in invertase activity. sds-page analysis of the co-purified enzymes suggests that the prote ...201424307498
n-glycosylation affects the proper folding, enzymatic characteristics and production of a fungal ß-glucosidase.heterologous expression of ß-glucosidase is one of the approaches to enhance the efficiency of fungal cellulase preparations. it has been reported that n-glycosylation affects the structure framework, function and stability of proteins. in this study, a ß-glucosidase from aspergillus terreus (genbank: xp_001216552, bgls) was heterologously expressed in pichia pastoris and trichoderma reesei. the four asparagine residues were all linked with high-mannose-type oligosaccharides in p. pastoris, wher ...201324308062
does a rhizospheric microorganism enhance k⁺ availability in agricultural soils?the potassium solubilizing microorganisms (ksms) are a rhizospheric microorganism which solubilizes the insoluble potassium (k) to soluble forms of k for plant growth and yield. k-solubilization is carried out by a large number of saprophytic bacteria (bacillus mucilaginosus, bacillus edaphicus, bacillus circulans, acidothiobacillus ferrooxidans, paenibacillus spp.) and fungal strains (aspergillus spp. and aspergillus terreus). major amounts of k containing minerals (muscovite, orthoclase, bioti ...201424315210
functional analysis of cis-aconitate decarboxylase and trans-aconitate metabolism in riboflavin-producing filamentous ashbya gossypii.in ashbya gossypii, isocitrate lyase (icl1) is a very crucial enzyme for riboflavin production. itaconate, the inhibitor of icl1, has been used as an antimetabolite for mutagenic studies in a. gossypii. it has been reported that itaconate is produced from cis-aconitate by cis-aconitate decarboxylase (cad1) in aspergillus terreus. in this study, identification of cad1 gene and determination of the presence of itaconate in the riboflavin biosynthetic pathway in a. gossypii were carried out to conf ...201424315530
new rubrolides from the marine-derived fungus aspergillus terreus oucmdz-1925.two new rubrolides, rubrolides r (1) and s (2), were isolated from the fermentation broth of the marine-derived fungus aspergillus terreus oucmdz-1925. their structures were elucidated on the basis of spectroscopic analysis and x-ray single crystal diffraction. compound 1 showed comparable or superior antioxidation against 2,2'-azino-di(3-ethylbenzthiazoline-6-sulfonic acid) (abts) radicals to those of trolox and ascorbic acid with an ic₅₀ value of 1.33 mm. compound 2 showed comparable or superi ...201424326339
disseminated aspergillosis in a dog due to aspergillus alabamensis.disseminated aspergillosis is uncommon in dogs and often associated with aspergillus terreus. a case of disseminated disease in an english springer spaniel is reported from which aspergillus alabamensis was recovered by culture and identified by molecular means suggesting a potential role for this agent as a primary pathogen of dogs.201224371723
aryl-aldehyde formation in fungal polyketides: discovery and characterization of a distinct biosynthetic mechanism.aryl-aldehydes are a common feature in fungal polyketides, which are considered to be exclusively generated by the r domain of nonreducing polyketide synthases (nr-pkss). however, by cloning and heterologous expression of both cryptic nr-pks and nonribosomal peptide synthase (nrps)-like genes from aspergillus terreus in saccharomyces cerevisiae, we identified a distinct mechanism for aryl-aldehyde formation in which a nrps-like protein activates and reduces an aryl-acid produced by the accompany ...201424412543
expression of the aspergillus terreus itaconic acid biosynthesis cluster in aspergillus niger.aspergillus terreus is a natural producer of itaconic acid and is currently used to produce itaconic acid on an industrial scale. the metabolic process for itaconic acid biosynthesis is very similar to the production of citric acid in aspergillus niger. however, a key enzyme in a. niger, cis-aconitate decarboxylase, is missing. the introduction of the a. terreus cada gene in a. niger exploits the high level of citric acid production (over 200 g per liter) and theoretically can lead to production ...201424438100
potential halophilic cellulases for in situ enzymatic saccharification of ionic liquids pretreated lignocelluloses.ionic liquids (ils) have been used as an alternative green solvent for lignocelluloses pretreatment. however, being a salt, ils exhibit an inhibitory effect on cellulases activity, thus making the subsequent saccharification inefficient. the aim of the present study is to produce salt-tolerant cellulases, with the rationale that the enzyme also tolerant to the presence of ils. the enzyme was produced from a locally isolated halophilic strain and was characterized and assessed for its tolerance t ...201424457303
an efficient tool for random insertional mutagenesis: agrobacterium tumefaciens-mediated transformation of the filamentous fungus aspergillus terreus.agrobacterium tumefaciens-mediated transformation (atmt) has been widely used in various organisms. in the current study, we developed a simple and efficient system for genetic transformation of the filamentous fungus aspergillus terreus using atmt. the transformation protocol was optimized for certain parameters to rapidly generate a library of transferred dna (t-dna) insertion mutants of a. terreus. the presence of mitotically stable hygromycin resistance gene (hph) integration in the genome w ...201424462974
crystal structure of an (r)-selective ω-transaminase from aspergillus terreus.chiral amines are important building blocks for the synthesis of pharmaceutical products, fine chemicals, and agrochemicals. ω-transaminases are able to directly synthesize enantiopure chiral amines by catalysing the transfer of an amino group from a primary amino donor to a carbonyl acceptor with pyridoxal 5'-phosphate (plp) as cofactor. in nature, (s)-selective amine transaminases are more abundant than the (r)-selective enzymes, and therefore more information concerning their structures is av ...201424498081
studies on extraction of mannanase enzyme by aspergillus terreus suk-1 from fermented palm kernel cake.microbial mannanases have become biotechnologically important in industry but their application is limited due to high production cost. in presents study, the extraction of mannanase from fermented palm kernel cake (pkc) in the solid state fermentation (ssf) was optimized. local isolate of aspergillus terreus suk-1 was grown on pkc in (ssf) using column bioreactor. the optimum condition were achieved after two washes of fermented pkc by adding of 10% glycerol (v/v) soaked for 10 h at the room te ...201324502150
optimization of complex fermentation media for glucose oxidase production using statistical approach.production cost of enzyme is largely determined by the type of the strain and raw material used to propagate the strain. hence, selection of the strain and raw materials is crucial in enzyme production. for glucose oxidase (gox), previous studies showed aspergillus terreus unimap aa-1 offers a better alternative to the existing sources. thus, a lower production cost could be logically anticipated by growing the strain in a cheaper complex media such as molasses. in this work, sugar cane molasses ...201324502155
optimization of inulinase production from low cost substrates using plackett-burman and taguchi methods.four marine-derived fungal isolates were screened for the production of inulinase enzyme from low cost substrates under solid state fermentation (ssf), one of them identified as aspergillus terreus showed the highest inulinase activity using artichoke leaves as a solid substrate. sequential optimization strategy, based on statistical experimental designs was employed to optimize the composition of the medium, including plackett-burman and taguchi's (l9 3(4)) orthogonal array designs. under the o ...201424507281
susceptibility breakpoints for amphotericin b and aspergillus species in an in vitro pharmacokinetic-pharmacodynamic model simulating free-drug concentrations in human serum.although conventional amphotericin b was for many years the drug of choice and remains an important agent against invasive aspergillosis, reliable susceptibility breakpoints are lacking. three clinical aspergillus isolates (aspergillus fumigatus, aspergillus flavus, and aspergillus terreus) were tested in an in vitro pharmacokinetic-pharmacodynamic model simulating the biphasic 24-h time-concentration profile of free amphotericin b concentrations in human serum with free peak concentrations (fcm ...201424514094
dissimilatory nitrate reduction by aspergillus terreus isolated from the seasonal oxygen minimum zone in the arabian sea.a wealth of microbial eukaryotes is adapted to life in oxygen-deficient marine environments. evidence is accumulating that some of these eukaryotes survive anoxia by employing dissimilatory nitrate reduction, a strategy that otherwise is widespread in prokaryotes. here, we report on the anaerobic nitrate metabolism of the fungus aspergillus terreus (isolate an-4) that was obtained from sediment in the seasonal oxygen minimum zone in the arabian sea, a globally important site of oceanic nitrogen ...201424517718
culture-based and sequence-based insights into biosynthesis of secondary metabolites by aspergillus terreus atcc 20542.aspergillus terreus atcc 20542 was cultivated in various culture media in order to activate its genome-encoded biosynthetic pathways and explore the secondary metabolic repertoire. in addition to mevinolinic acid (lovastatin) and its precursor monacolin l, a number of other secondary metabolites were found in the analyzed cultures, namely terreic acid, citrinin, (+)-geodin, terrein, and dehydrocurvularin. in contrast to previously described gene clusters responsible for production of lovastatin, ...201424534845
antibiotic-producing properties of aspergillus terreus. 195424543173
expression, purification, crystallization and preliminary x-ray diffraction analysis of aspergillus terreus endo-β-1,4-glucanase from glycoside hydrolase family 12.endoglucanases are important enzymes that are involved in the modification and degradation of cellulose. filamentous fungi such as aspergillus terreus are effective biomass degraders in nature owing to their capacity to produce an enzymatic arsenal of glycoside hydrolases, including endoglucanase from glycoside hydrolase family 12 (gh12). the a. terreus gh12 endoglucanase was cloned and overexpressed in a. nidulans, purified and crystallized. a single crystal was obtained from a solution consist ...201424637772
fermented whey as poultry feed additive to prevent fungal contamination.fungal contamination of poultry feed causes economic losses to industry and represents a potential risk to animal health. the aim of the present study was to analyze the effectiveness of whey fermented with kefir grains as additive to reduce fungal incidence, thus improving feed safety.201424652751
upstream and downstream processing of lovastatin by aspergillus terreus.the present study describes the enhanced production and purification of lovastatin by aspergillus terreus in submerged batch fermentation. the enhancement of lovastatin production from a. terreus was attempted by random mutagenesis using ultraviolet radiations and nitrous acid. uv mutants exhibited increased efficiency for lovastatin production as compared with nitrous acid mutants. among all the mutants developed, a. terreus uv-4 was found to be the hyper producer of lovastatin. this mutant gav ...201424671671
effect of mineral constituents in the bioleaching of uranium from uraniferous sedimentary rock samples, southwestern sinai, egypt.bioleaching, like biotechnology uses microorganisms to extract metals from their ore materials, whereas microbial activity has an appreciable effect on the dissolution of toxic metals and radionuclides. bioleaching of uranium was carried out with isolated fungi from uraniferous sedimentary rocks from southwestern sinai, egypt. eight fungal species were isolated from different grades of uraniferous samples. the bio-dissolution experiments showed that aspergillus niger and aspergillus terreus exhi ...201424682031
optimization of xylanase production by filamentous fungi in solid-state fermentation and scale-up to horizontal tube bioreactor.five microorganisms, namely aspergillus niger cect 2700, a. niger cect 2915, a. niger cect 2088, aspergillus terreus cect 2808, and rhizopus stolonifer cect 2344, were grown on corncob to produce cell wall polysaccharide-degrading enzymes, mainly xylanases, by solid-state fermentation (ssf). a. niger cect 2700 produced the highest amount of xylanases of 504 ± 7 u/g dry corncob (dcc) after 3 days of fermentation. the optimization of the culture broth (5.0 g/l nano3, 1.3 g/l (nh4)2so4, 4.5 g/l kh2 ...201424728763
filamentous fungi in microtiter plates-an easy way to optimize itaconic acid production with aspergillus terreus.itaconic acid is an important industrial building block and is produced by the filamentous fungi aspergillus terreus. to make the optimization process more efficient, a scale-down from shake flasks to microtiter plates was performed. this resulted in comparable product formations, and 87.7 g/l itaconic acid was formed after 10 days of cultivation in the microtiter plate. the components of the minimal medium were varied independently for a media optimization. this resulted in an increase of the i ...201424737061
filamentous fungal diversity and community structure associated with the solid state fermentation of chinese maotai-flavor liquor.maotai-flavor liquor is produced by simultaneous saccharification and fermentation (ssf) process under solid state conditions, including daqu (starter) making, stacking fermentation and alcohol fermentation stages. filamentous fungi produce many enzymes to degrade the starch material into fermentable sugar during liquor fermentation. this study investigated the filamentous fungal community associated with liquor making process. eight and seven different fungal species were identified by using cu ...201424742997
molecular characterization and expression of a novel alcohol oxidase from aspergillus terreus mtcc6324.the alcohol oxidase (aox) cdna from aspergillus terreus mtcc6324 with an open reading frame (orf) of 2001 bp was constructed from n-hexadecane induced cells and expressed in escherichia coli with a yield of ∼4.2 mg protein g-1 wet cell. the deduced amino acid sequences of recombinant raox showed maximum structural homology with the chain b of aryl aox from pleurotus eryngii. a functionally active aox was achieved by incubating the apo-aox with flavin adenine dinucleotide (fad) for ∼80 h at 16°c ...201424752075
terrein biosynthesis in aspergillus terreus and its impact on phytotoxicity.terrein is a fungal metabolite with ecological, antimicrobial, antiproliferative, and antioxidative activities. although it is produced by aspergillus terreus as one of its major secondary metabolites, not much is known about its biosynthetic pathway. here, we describe an unexpected discovery of the terrein biosynthesis gene locus made while we were looking for a pks gene involved in production of conidia coloration pigments common for aspergilli. the gene, ateg_00145, here named tera, is essent ...201424816227
does fungicide application in vineyards induce resistance to medical azoles in aspergillus species?this study assessed if the use of sterol demethylase inhibitor fungicides in vineyard production can induce resistance to azoles in aspergillus strains and if it can induce selection of resistant species. we also tried to identify the aspergillus species most prevalent in the vineyards. two vineyards from northern portugal were selected from "vinhos verdes" and "douro" regions. the vineyards were divided into plots that were treated or not with penconazole (pen). in each vineyard, air, soil, and ...201424833021
altering the activation mechanism in thermomyces lanuginosus lipase.it is shown by rational site-directed mutagenesis of the lid region in thermomyces lanuginosus lipase that it is possible to generate lipase variants with attractive features, e.g., high lipase activity, fast activation at the lipid interface, ability to act on water-soluble substrates, and enhanced calcium independence. the rational design was based on the lid residue composition in aspergillus niger ferulic acid esterase (faea). five constructs included lipase variants containing the full faea ...201424870718
screening of indigenously isolated fungi for lovastatin production and its in vivo evaluation.seven indigenously isolated fungal strains (aspergillus niger, aspergillus flavus, aspergillus terreus, penicillium citrinum, penicillium notatum, pleurotus ostreatus and trichoderma viradae) were tested for their potential to produce cholesterol lowering drug lovastatin by using different agro-industrial wastes (corn cobs, corn stover, banana stalk, wheat straw, wheat bran, bagasse) in submerged as well as solid state fermentation. aspergillus terreus showed maximum production of 18.74 mg/100 m ...201424894549
invasion of solanum tuberosum l. by aspergillus terreus: a microscopic and proteomics insight on pathogenicity.aspergillus terreus is one of the most harmful filamentous fungal pathogen of humans, animals and plants. recently, researchers have discovered that a. terreus can cause foliar blight disease in potato (solanum tuberosum l.). we used light and scanning electron microscopy, and performed proteomics analysis in an attempt to dissect the invasion process of a. terreus in this important crop.201424917207
effect of salt on survival and p-solubilization potential of phosphate solubilizing microorganisms from salt affected soils.a total of 23 phosphate solubilizing bacteria (psb) and 35 phosphate solubilizing fungi (psf) were isolated from 19 samples of salt affected soils. the ability of 12 selected psb and psf to grow and solubilize tricalcium phosphate in the presence of different concentrations of nacl was examined. among 12 psb, aerococcus sp. strain psbcrg1-1 recorded the highest (12.15) log viable cell count at 0.4 m nacl concentration after 7 days after incubation (dai) and the lowest log cell count (1.39) was r ...201224936136
enhancement of in vitro high-density polyethylene (hdpe) degradation by physical, chemical, and biological treatments.partially degraded high-density polyethylene (hdpe) was collected from plastic waste dump yard for biodegradation using fungi. of various fungi screened, strain mf12 was found efficient in degrading hdpe by weight loss and fourier transform infrared (ft-ir) spectrophotometric analysis. strain mf12 was selected as efficient hdpe degraders for further studies, and their growth medium composition was optimized. among those different media used, basal minimal medium (bmm) was suitable for the hdpe d ...201424946709
promiscuity breeds diversity: the role of polyketide synthase in natural product biosynthesis.secondary metabolites are often highly biologically active molecules and are widely used from antibacterial to anticancer drugs. in this issue of chemistry and biology, zaehle and coworkers describe the gene cluster and biosynthesis of the polyketide terrein, a secondary metabolite produced by the soil-borne fungus aspergillus terreus.201424950100
identification and characterisation of organisms associated with chocolate pralines and sugar syrups used for their production.spoilage of chocolate pralines, due to growth of microorganisms tolerating low water activity, causes problems in the confectionary industry. therefore, an increased knowledge on which organisms are present in the chocolate fillings and their tolerance towards low aw, ph, ethanol and other preservatives is needed. using media containing 40-50% glucose (aw 0.872-0.925) bacteria, yeasts and moulds were isolated from chocolate pralines (aw 0.70-0.898) of nine manufactures and sugar syrups (aw 0.854 ...201424997272
aspergillus terreus recovered from a corneal scraping.a 52 year old, healthy male presented to his optometrist complaining of redness and irritation in the right eye. a foreign body was removed from the eye. the patient was started on ophthalmic solutions of vigamox and systane. at 48 hours, the patient reported increased redness, limited vision, and yellow discharge from the eye. the patient was referred to an ophthalmologist for further evaluation. physical assessment revealed a superlative central infiltrate (extreme, centrally located injury th ...201425000648
invasive aspergillosis caused by aspergillus terreus: an emerging opportunistic infection with poor outcome independent of azole therapy.invasive aspergillosis (ia) caused by aspergillus terreus is a significant cause of morbidity and mortality in patients with haematological malignancy (hm). very few data are available in this patient population to differentiate ia patients with a. terreus from those with non-terreus species of aspergillus to compare outcomes. we retrospectively investigated 513 hm patients who were treated for either definite or probable ia between june 1993 and august 2012 in a cancer centre.201425006241
efficient expression, purification, and characterization of a novel fad-dependent glucose dehydrogenase from aspergillus terreus in pichia pastoris.flavin adenine dinucleotide-dependent glucose dehydrogenase (fad-gdh) can utilize a variety of external electron acceptors and also has stricter substrate specificity than any other glucose oxidoreductases, which makes it the ideal diagnostic enzyme in the field of glucose biosensors. a gene coding for a hypothetical protein, similar to glucose oxidase and derived from aspergillus terreus nih2624, was overexpressed in pichia pastoris gs115 under the control of an aox1 promoter with a level of 26 ...201425022525
optimization of β-glucosidase production by aspergillus terreus strain emoo 6-4 using response surface methodology under solid-state fermentation.forty-two morphologically different fungal strains were isolated from different soil samples and agricultural wastes and screened for β-glucosidase activity under solid-state fermentation. eight species were chosen as the most active β-glucosidase producers and were subjected to primary morphological identification. β-glucosidase was highly produced by aspergillus terreus, which showed the highest activity, and was subjected to full identification using scanning electron microscopy and molecular ...201525036937
cellulolytic and xylanolytic enzymes from thermophilic aspergillus terreus rwy.thermophilic aspergillus terreus rwy produced cellulases and xylanases in optimal concentrations at 45 °c in solid state fermentation process, though enzyme production was also observed at 50 and 55 °c. filter paper cellulase (fp), endoglucanase (eg), β-glucosidase (bgl), cellobiohydrolase (cbh), xylanase, β-xylosidase, α-l-arabinofuranosidase and xylan esterase activities for a. terreus rwy at 45 °c in 72 h were 11.3 ± 0.65, 103 ± 6.4, 122.5 ± 8.7, 10.3 ± 0.66, 872 ± 22.5, 22.1 ± 0.75, 126.4 ±  ...201425047723
comparative study of toxicity of azo dye procion red mx-5b following biosorption and biodegradation treatments with the fungi aspergillus niger and aspergillus terreus.azo dyes are an important class of environmental contaminants and are characterized by the presence of one or more azo bonds (-n=n-) in their molecular structure. effluents containing these compounds resist many types of treatments due to their molecular complexity. therefore, alternative treatments, such as biosorption and biodegradation, have been widely studied to solve the problems caused by these substances, such as their harmful effects on the environment and organisms. the aim of the pres ...201425048922
c7-prenylation of tryptophanyl and o-prenylation of tyrosyl residues in dipeptides by an aspergillus terreus prenyltransferase.during our search for novel prenyltransferases, a putative gene ateg_04218 from aspergillus terreus raised our attention and was therefore amplified from strain dsm 1958 and expressed in escherichia coli. biochemical investigations with the purified recombinant protein and different aromatic substrates in the presence of dimethylallyl diphosphate revealed the acceptance of all the tested tryptophan-containing cyclic dipeptides. structure elucidation of the main enzyme products by nmr and ms anal ...201525125042
in vitro combination of isavuconazole with micafungin or amphotericin b deoxycholate against medically important molds.whether isavuconazole, an extended-spectrum triazole, possesses synergistic activity in combination therapy with echinocandins or amphotericin b for the treatment of invasive molds infections has not been studied. our in vitro combination studies showed that isavuconazole and micafungin are synergistically active against aspergillus fumigatus, aspergillus flavus, aspergillus terreus, and cunninghamella bertholletiae. these results suggest that isavuconazole, in combination with micafungin, may h ...201425136021
synthetic (+)-terrein suppresses interleukin-6/soluble interleukin-6 receptor induced-secretion of vascular endothelial growth factor in human gingival fibroblasts.interleukin (il)-6 is a proinflammatory cytokine that performs a wide variety of biological functions, including important roles in the progression of chronic inflammatory diseases such as periodontal disease. (+)-terrein, a secondary bioactive fungal metabolite isolated from aspergillus terreus, has various biological activities; however, its anti-inflammatory effects are still unknown. the purpose of this study was to examine the effect of synthetic (+)-terrein on il-6 signaling and related pr ...201425151086
successful surgical treatment of rare aspergillus terreus prosthetic valve endocarditis complicated by intracranial and mesenteric artery mycotic aneurysms. 201425156465
direct production of itaconic acid from liquefied corn starch by genetically engineered aspergillus terreus.itaconic acid is on the doe (department of energy) top 12 list of biotechnologically produced building block chemicals and is produced commercially by aspergillus terreus. however, the production cost of itaconic acid is too high to be economically competitive with the petrochemical-based products. itaconic acid is generally produced from raw corn starch, including three steps: enzymatic hydrolysis of corn starch into a glucose-rich syrup by α-amylase and glucoamylase, fermentation, and recovery ...201425162619
improving itaconic acid production through genetic engineering of an industrial aspergillus terreus strain.itaconic acid, which has been declared to be one of the most promising and flexible building blocks, is currently used as monomer or co-monomer in the polymer industry, and produced commercially by aspergillus terreus. however, the production level of itaconic acid hasn't been improved in the past 40 years, and mutagenesis is still the main strategy to improve itaconate productivity. the genetic engineering approach hasn't been applied in industrial a. terreus strains to increase itaconic acid p ...201425162789
treatment of aspergillus terreus infections: a clinical problem not yet resolved.despite the use of recommended therapies, invasive infections by aspergillus terreus show a poor response. for years, investigative studies on the failure of therapy of fungal infections have focused on in vitro susceptibility data. however, it is well known that low minimum inhibitory concentrations (mics) are not always predictive of response to therapy despite a correct dosage schedule. many experimental and clinical studies have tried to establish a relationship between mics and outcome in s ...201425190543
novel fungal consortium for bioremediation of metals and dyes from mixed waste stream.the present study is targeted towards development of a three member fungal consortium for effective removal of metals [cr(6+) and cu(2+)] and dyes [ab and po] from mixed waste streams. initial studies using individual fungal strain showed that aspergillus lentulus was best for cu(2+) and ab removal, aspergillus terreus for cr(6+) removal whereas, rhizopus oryzae was best for po removal. based on the complementary pollutant affinities and positive interactions, a consortium comprising all three s ...201425203229
influence of the ph on the itaconic acid production with aspergillus terreus.itaconic acid is mainly produced with the filamentous fungi aspergillus terreus. an increase in the ph during the production phase of the cultivation resulted in an increase in the itaconic acid concentration. the ph was raised by a single ph shift ranging from ph 4 to 6 or by a ph control to ph 3. different lyes can be used for the ph shift, but ammonia solution has proven to be the best, because here the productivity does not drop after the ph shift. the highest itaconic acid concentration of ...201425213913
aspergillus terreus endogenous endophthalmitis: report of a case and review of literature.we report a rare case of aspergillus terreus endogenous endophthalmitis in an immunocompetent patient with subretinal abscess and also review the reported cases. a 50-year-old healthy male presented with sudden painful loss of vision in right eye. he was diagnosed with endogenous endophthalmitis and underwent urgent vitrectomy. aspergillus terreus growth was obtained in culture. at final follow-up, there was complete resolution of the infection but visual acuity was poor due to macular scar. asp ...201425230968
novel and sensitive qpcr assays for the detection and identification of aspergillosis causing species.despite concerted efforts, diagnosis of aspergillosis is still a great challenge to clinical microbiology laboratories. along with the requirement for high sensitivity and specificity, species-specific identification is important. we developed rapid, sensitive and species-specific qpcr assays using the taqman technology for the detection and identification of aspergillus fumigatus and aspergillus terreus. the assays were designed to target orthologs of the streptomyces factor c gene that are onl ...201425261942
molecular genetic characterization of terreic acid pathway in aspergillus terreus.terreic acid is a natural product derived from 6-methylsalicylic acid (6-msa). a compact gene cluster for its biosynthesis was characterized. isolation of the intermediates and shunt products from the mutant strains, combined with bioinformatic analyses, allowed for the proposition of a biosynthetic pathway for terreic acid.201425265334
metabolic engineering of itaconate production in escherichia coli.interest in sustainable development has led to efforts to replace petrochemical-based monomers with biomass-based ones. itaconic acid, a c5-dicarboxylic acid, is a potential monomer for the chemical industry with many prospective applications. cis-aconitate decarboxylase (cada) is the key enzyme of itaconate production, converting the citric acid cycle intermediate cis-aconitate into itaconate. heterologous expression of cada from aspergillus terreus in escherichia coli resulted in low cada acti ...201525277412
the marine-derived fungal metabolite, terrein, inhibits cell proliferation and induces cell cycle arrest in human ovarian cancer cells.the difficulties faced in the effective treatment of ovarian cancer are multifactorial, but are mainly associated with relapse and drug resistance. cancer stem-like cells have been reported to be an important contributor to these hindering factors. in this study, we aimed to investigate the anticancer activities of a bioactive fungal metabolite, namely terrein, against the human epithelial ovarian cancer cell line, skov3, primary human ovarian cancer cells and ovarian cancer stem-like cells. ter ...201425318762
fungal production of single cell oil using untreated copra cake and evaluation of its fuel properties for biodiesel.this study evaluated the microbial conversion of coconut oil waste, a major agro-residue in tropical countries, into single cell oil (sco) feedstock for biodiesel production. copra cake was used as a low-cost renewable substrate without any prior chemical or enzymatic pretreatment for submerged growth of an oleaginous tropical mangrove fungus, aspergillus terreus ibb m1. the sco extracted from fermented biomass was converted into fatty acid methyl esters (fames) by transesterification and evalua ...201525341469
invasive aspergillosis in liver transplant recipients: epidemiology, clinical characteristics, treatment, and outcomes in 116 cases.invasive aspergillosis (ia) in liver transplant recipients is associated with grave outcomes. we reviewed 116 individual cases reported in the literature from 1985 to 2013. ia was diagnosed after a median of 25 days after transplantation and involved a single organ in 51% of the cases, whereas in the remaining cases, multiple sites were involved. the most common infecting aspergillus species were aspergillus fumigatus (73%), aspergillus flavus (14%), and aspergillus terreus (8%). amphotericin b ...201525348192
aspergillus terreus infection in a sutureless self-sealing incision made during cataract surgery.here, we describe a case of keratitis caused by aspergillus terreus in an 80-year-old immunocompetent woman 1 month after uneventful cataract surgery. the patient presented with decreased visual acuity (20/50) and severe pain in her right eye. examination revealed a 3.5 × 4.5 mm white-colored deep stromal infiltration located on the temporal corneal tunnel incision. corneal scraping samples were obtained for cytological and culture examinations. the cinnamon-brown colonies consisting of a dense ...201525362536
aspergillus fumigatus and related species.the genus aspergillus contains etiologic agents of aspergillosis. the clinical manifestations of the disease range from allergic reaction to invasive pulmonary infection. among the pathogenic aspergilli, aspergillus fumigatus is most ubiquitous in the environment and is the major cause of the disease, followed by aspergillus flavus, aspergillus niger, aspergillus terreus, aspergillus nidulans, and several species in the section fumigati that morphologically resemble a. fumigatus. patients that a ...201425377144
the overproduction of 2,4-dtbp accompanying to the lack of available form of phosphorus during the biodegradative utilization of aminophosphonates by aspergillus terreus.although information about the ability of some filamentous fungi to biodegrade organophosphonates is available, the knowledge about accompanying changes in fungal metabolism is very limited. the aim of our study was to determine the utilization of the chosen, structurally diverse aminophosphonates by aspergillus terreus (thom), in the context of the behaviour of this fungus while growing in unfavourable conditions, namely the lack of easily available phosphates. we found that all the studied com ...201525385070
expression, characterization and mutagenesis of an fad-dependent glucose dehydrogenase from aspergillus terreus.an fad-dependent glucose dehydrogenase (fad-gdh) from aspergillus terreus nih2624 was expressed in escherichia coli with a yield of 228±16u/l of culture. co-expression with chaperones dnak/dnaj/grpe and osmotic stress induced by simple carbon sources enhanced productivity significantly, improving the yield to 23883±563u/l after optimization. fad-gdh was purified in two steps with the specific activity of 604u/mg. using d-glucose as substrate, the optimal ph and temperature for fad-gdh were deter ...201525435504
reactive oxygen species regulate lovastatin biosynthesis in aspergillus terreus during submerged and solid-state fermentations.in a previous work we detected an important increase in reactive oxygen species (ros) concentrations during idiophase in lovastatin fermentations. hence, the objective of the present work was to determine if ros contributes to the regulation of lovastatin biosynthesis. exogenous antioxidants were used to reduce ros accumulation. the addition of n-acetyl-l-cysteine (nac) decreased ros accumulation and concurrent lovastatin production. in solid-state fermentation (ssf), the addition of 100 mm of n ...201425457945
molecular epidemiology of aspergillus collected from cystic fibrosis patients.aspergillus respiratory infection is a common complication in cystic fibrosis (cf) and is associated with loss of pulmonary function and allergic disease.201525459562
production of itaconic acid in escherichia coli expressing recombinant α-amylase using starch as substrate.several studies on fermentative production of a vinyl monomer itaconic acid from hydrolyzed starch using aspergillus terreus have been reported. herein, we report itaconic acid production by escherichia coli expressing recombinant α-amylase, using soluble starch as its sole carbon source. to express α-amylase in e. coli, we first constructed recombinant plasmids expressing α-amylases by using cell surface display technology derived from two amylolytic bacteria, bacillus amyloliquefaciens nbrc 15 ...201525468427
pathway transfer in fungi.itaconic acid is an important building block for the chemical industry. currently, aspergillus terreus is the main organism used for itaconic acid production. due to the enormous citric acid production capacity of aspergillus niger, this host is investigated as a potential itaconic acid production host. several strategies have been tried so far: fermentation optimization, expression of cis-aconitate decarboxylase (cada) alone and in combination with aconitase targeted to the same compartment, ch ...201725482236
immobilization of aspergillus terreus lipase in self-assembled hollow nanospheres for enantioselective hydrolysis of ketoprofen vinyl ester.the aim of this study was to improve the ability of aspergillus terreus lipase to separate the racemic ketoprofen vinyl ester into individual enantiomers using hollow self-assembly alginate-graft-poly(ethylene glycol)/α-cyclodextrins (alg-g-peg/α-cd) spheres as enzyme immobilization carriers. the morphology and size of the alg-g-peg/α-cd particles were investigated by transmission electron microscopy (tem) and were found to be nanoscale. to facilitate recycling, calcium alginate (ca) beads were ...201525483320
aspergillus citrinoterreus, a new species of section terrei isolated from samples of patients with nonhematological predisposing conditions.the use of molecular identification techniques has revealed an increasing number of new species within aspergillus section terrei. we phenotyped a set of 26 clinical isolates that showed genetic differences from aspergillus terreus sensu stricto by analyzing sequences from pcr-amplified β-tubulin and calmodulin genes and the internal transcribed spacer region. since the isolates were phylogenetically and morphologically different from all of the members of aspergillus section terrei, they are de ...201525502530
territrem and butyrolactone derivatives from a marine-derived fungus aspergillus terreus.seventeen lactones including eight territrem derivatives (1-8) and nine butyrolactone derivatives (9-17) were isolated from a marine-derived fungus aspergillus terreus scsgaf0162 under solid-state fermentation of rice. compounds 1-3 and 9-10 were new, and their structures were elucidated by spectroscopic analysis. the acetylcholinesterase inhibitory activity and antiviral activity of compounds 1-17 were evaluated. among them, compounds 1 and 2 showed strong inhibitory activity against acetylchol ...201425522319
effect of reduced oxygen on the antifungal susceptibility of clinically relevant aspergilli.the influence of hypoxia on the in vitro activities of amphotericin b, azoles, and echinocandins against aspergillus spp. was evaluated by comparing mics, minimal fungicidal concentrations (mfcs), and epidemiological cutoffs (ecoffs). changes of mic distributions due to hypoxia largely depend on the method, the species, and the growth ability under hypoxia. the activities of antifungals were not significantly altered under hypoxia, except for aspergillus terreus, for which the activity changed f ...201525547350
simultaneous saccharification and fermentation of delignified lignocellulosic biomass at high solid loadings by a newly isolated thermotolerant kluyveromyces sp. for ethanol production.simultaneous saccharification and fermentation studies were carried out using thermotolerant newly isolated kluyveromyces sp. with three different delignified lignocellulosic biomass viz. rice straw, wheat straw and sugarcane bagasse at 5-15% solid loading and 6-12 fpu g(-1) substrate enzyme loading for different time intervals 0-72 h at 42°c. maximum ethanol achieved from rice straw, wheat straw and sugarcane bagasse with in-house crude cellulases from aspergillus terreus was 23.23, 18.29 and 1 ...201525553563
antifungal activity of compounds targeting the hsp90-calcineurin pathway against various mould species.invasive mould infections are associated with a high mortality rate and the emergence of mdr moulds is of particular concern. calcineurin and its chaperone, the heat shock protein 90 (hsp90), represent an important pathway for fungal virulence that can be targeted at different levels. we investigated the antifungal activity of compounds directly or indirectly targeting the hsp90-calcineurin axis against different mould species.201525558076
[superficial mycoses in patients from anzoátegui state, venezuela, period 2002-2012].superficial fungal diseases that affect the skin and its appendages are frequently seen in basic triage and in dermatology services. these diseases are distributed in venezuela with an incidence of 92.9%. the aim of this study was to determine the genera and species that cause dermatomycoses in residents of anzoátegui state, venezuela, during the period 2002-2012. a total of 4257 patients with a presumptive diagnosis of superficial mycoses were studied, with ages from 7 months to 79 years. the o ...201425558751
recent advances in genome mining of secondary metabolites in aspergillus terreus.filamentous fungi are rich resources of secondary metabolites (sms) with a variety of interesting biological activities. recent advances in genome sequencing and techniques in genetic manipulation have enabled researchers to study the biosynthetic genes of these sms. aspergillus terreus is the well-known producer of lovastatin, a cholesterol-lowering drug. this fungus also produces other sms, including acetylaranotin, butyrolactones, and territram, with interesting bioactivities. this review wil ...201425566227
anticancer activity of fungal l-asparaginase conjugated with zinc oxide nanoparticles.demand for developing novel delivery system for cancer treatment has increased due to the side effects present in intravenous injection of l-asparaginase. nanoparticles are used for delivering the drugs to its destination in cancer cure. nanobiocomposite of zinc oxide nanoparticles conjugated with l-asparaginase was produced by aspergillus terreus and was confirmed using maximum uv-vis absorption at 340 nm in the present work. the presence of functional groups like oh, c-h, -c=n and c=o on the s ...201525589205
onychomycosis in israel: epidemiological aspects.onychomycosis is a fungal infection treated orally for prolonged periods of treatment, caused primarily by dermatophytes, candida species and non-dermatophyte moulds (ndms). the prevalence of specific aetiology may differ in dependence of environmental, geographic and demographic factors, and may affect management of the infection. the objective of this survey was to analyse epidemiologic parameters of onychomycosis in israel. data of a cohort of 27,093 patients were collected from six centres d ...201525590832
(+)‑terrein inhibits human hepatoma bel‑7402 proliferation through cell cycle arrest.hepatoma is a common malignant tumor. thus, the development of a high‑efficacy therapeutic drug for hepatoma is required. in this study, (+)‑terrein isolated from the marine sponge‑derived aspergillus terreus pf‑26 against cell growth, apoptosis and cell cycle were assessed by mtt and flow cytometry. mrna array containing 73 cell cycle‑related genes and three cell morphology‑related genes was generated and its performance evaluated. the cell cycle pathway map was created using the pathview packa ...201525592371
subcutaneous mycoses: an aetiological study of 15 cases in a tertiary care hospital at dibrugarh, assam, northeast india.subcutaneous mycoses are a group of fungal infections of dermis and subcutaneous tissue which consist of sporotrichosis, chromoblastomycosis, phaeohyphomycosis, hyalohyphomycosis, mycetoma, subcutaneous zygomycosis, rhinosporidiosis, lobomycosis and disseminated penicilliosis. a total of 46 consecutive patients with clinically suspected subcutaneous mycoses attending various departments of assam medical college and hospital were included in this prospective study to know the prevalence of subcut ...201525596859
the cell factory aspergillus enters the big data era: opportunities and challenges for optimising product formation.living with limits. getting more from less. producing commodities and high-value products from renewable resources including waste. what is the driving force and quintessence of bioeconomy outlines the lifestyle and product portfolio of aspergillus, a saprophytic genus, to which some of the top-performing microbial cell factories belong: aspergillus niger, aspergillus oryzae and aspergillus terreus. what makes them so interesting for exploitation in biotechnology and how can they help us to addr ...201525616499
influence of oxygen on lovastatin biosynthesis by aspergillus terreus atcc 20542 quantitatively studied on the level of individual pellets.despite oxygen is believed to be the most important environmental factor for any aerobic microbial process, the quantitative studies of its influence on growth and metabolite formation on the level of individual pellets formed by filamentous fungi were seldom performed. never was it made for lovastatin producer aspergillus terreus atcc20542. thus, this work is a quantitative study of oxygen transfer into a. terreus pellets during lovastatin biosynthesis in the shake flask culture. the basic meas ...201525627471
bioprocess engineering aspects of the cultivation of a lovastatin producer aspergillus terreus.the aim of this work is to review bioprocess engineering aspects of lovastatin (antihypercholesterolemia drug) production by aspergillus terreus in the submerged culture in the bioreactors of various scale presented in the scientific literature since the nineties of the twentieth century. the key factor influencing the cultivation of any filamentous species is fungal morphology and that is why this aspect was treated as the starting point for further considerations. fungal morphology is known to ...201525633258
evaluation of antifungal combinations of nystatin-intralipid against aspergillus terreus using checkerboard and disk diffusion methods.the objective of this study was to evaluate the efficacy of combinations of nystatin-intralipid, found previously to be more active than nystatin, with antifungals of different mode of activity, against aspergillus terreus.201525639922
aspergillus species as emerging causative agents of onychomycosis.onychomycosis is a common nail infection caused by dermatophytes, non-dermatophyte molds (ndm), and yeasts. aspergillus species are emerging as increasing causes of toenail onychomycosis. the purpose of this study was species delineation of aspergillus spp. isolated from patients with onychomycosis.201525656408
uncovering the unusual d-ring construction in terretonin biosynthesis by collaboration of a multifunctional cytochrome p450 and a unique isomerase.terretonin (1) is a fungal meroterpenoid isolated from aspergillus terreus, and possesses a highly oxygenated and unique tetracyclic structure. although the biosynthetic gene cluster for 1 has been identified and the biosynthesis has recently been studied by heterologous reconstitution and targeted-gene deletion experiments, the last few steps of the terretonin pathway after terrenoid (6) have yet to be elucidated. notably, the mechanism for the d-ring expansion to afford the terretonin scaffold ...201525671343
production of plant cell wall degrading enzymes by monoculture and co-culture of aspergillus niger and aspergillus terreus under ssf of banana peels.filamentous fungi are considered to be the most important group of microorganisms for the production of plant cell wall degrading enzymes (cwde), in solid state fermentations. in this study, two fungal strains aspergillus niger ms23 and aspergillus terreus ms105 were screened for plant cwde such as amylase, pectinase, xylanase and cellulases (β-glucosidase, endoglucanase and filterpaperase) using a novel substrate, banana peels (bp) for ssf process. this is the first study, to the best of our kn ...201425763058
lumazine peptides from the marine-derived fungus aspergillus terreus.terrelumamides a (1) and b (2), two new lumazine-containing peptides, were isolated from the culture broth of the marine-derived fungus aspergillus terreus. from the results of combined spectroscopic and chemical analyses, the structures of these compounds were determined to be linear assemblies of 1-methyllumazine-6-carboxylic acid, an amino acid residue and anthranilic acid methyl ester connected by peptide bonds. these new compounds exhibited pharmacological activity by improving insulin sens ...201525775424
molecular epidemiology and in-vitro antifungal susceptibility of aspergillus terreus species complex isolates in delhi, india: evidence of genetic diversity by amplified fragment length polymorphism and microsatellite typing.aspergillus terreus is emerging as an etiologic agent of invasive aspergillosis in immunocompromised individuals in several medical centers in the world. infections due to a. terreus are of concern due to its resistance to amphotericin b, in vivo and in vitro, resulting in poor response to antifungal therapy and high mortality. herein we examined a large collection of molecularly characterized, geographically diverse a. terreus isolates (n = 140) from clinical and environmental sources in india ...201525781896
in vitro activities of amphotericin b, terbinafine, and azole drugs against clinical and environmental isolates of aspergillus terreus sensu stricto.the antifungal susceptibilities of 40 clinical and environmental isolates of a. terreus sensu stricto to amphotericin b, terbinafine, itraconazole, and voriconazole were determined in accordance with clsi document m38-a2. all isolates had itraconazole and voriconazole mics lower than epidemiologic cutoff values, and 5% of the isolates had amphotericin b mics higher than epidemiologic cutoff values. terbinafine showed the lowest mics. no significant differences were found when mics of clinical an ...201525824228
purification and biochemical characterization of an extracellular serine peptidase from aspergillus terreus.peptidases are important because they play a central role in pharmaceutical, food, environmental, and other industrial processes. a serine peptidase from aspergillus terreus was isolated after two chromatography steps that showed a yield of 15.5%. its molecular mass was determined to be 43 kd, by sodium dodecyl sulfate polyacrylamide gel electrophoresis (sds-page). this peptidase was active between ph 5.0 to 8.0 and had maximum activity at ph 7.0, at 45°c. when exposited with 1 m of urea, the en ...201625830777
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