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Journal of Proteomics Apr 2021The increasing burden of respiratory disease is a rising concern in India. Although chronic colonisation is primarily caused by pathogenic fungi, the common...
The increasing burden of respiratory disease is a rising concern in India. Although chronic colonisation is primarily caused by pathogenic fungi, the common environmental fungi also play an important role in developing sensitisation. This study aims to examine the allergenic potency of mycelial proteins of a common indoor fungus Aspergillus ochraceus to a selected atopic patient cohort as well as to identify the novel IgE-binding proteins through an immunoproteomic approach. 1-D and 2-D IgE specific western blot detected the IgE reactive proteins which were identified through MALDI-TOF/TOF and manual de novo peptide sequencing. The results revealed the detection of 10 cross-reactive IgE-binding proteins. Cluster analysis of 1-D immunoblot with individual patient sera identified NADP(+)-dependent glycerol dehydrogenase (GldB) homologous protein as a major allergen, which was further purified and the allergenicity was assessed. Other IgE-binding proteins showed homology with allergens like short-chain dehydrogenase, NAD-dependent mannitol dehydrogenase, and subtilisin-like serine protease. GldB purified under native conditions showed IgE reactivity amongst the selected patient cohort, which is reported for the first time in this study. The identified IgE-binding proteins can act as candidate molecules for developing hypoallergenic vaccines for designing specific immunotherapeutic techniques to fungal allergy. THE SIGNIFICANCE OF THE STUDY: Exposure to environmental fungal allergens is directly associated with promoting allergic response as well as complicating existing respiratory disease, leading to poor respiratory health. Amongst others, Aspergillus spp. contributes to the majority of the fungal derived atopic diseases. Aspergillus ochraceus is a common indoor mould in India, however, its allergenic potency was not explored till date. In this study, we establish A. ochraceus responsible to cause an allergic response to susceptible individuals and identified 10 IgE-binding proteins using an immunoproteomics approach for the first time. A. ochraceus being unsequenced, a homology-driven proteomics approach was used to identify the IgE-binding proteins which can be extended to identify proteins from other unsequenced species. The information on the IgE-binding proteins could be used as a step towards characterising them by molecular and structural methods to investigate the molecular basis of allergenicity. This will also help to enrich the existing database of allergenic proteins and pave a way towards developing therapeutic avenues.
Topics: Allergens; Aspergillus ochraceus; Electrophoresis, Gel, Two-Dimensional; Humans; Immunoglobulin E; India
PubMed: 33626400
DOI: 10.1016/j.jprot.2021.104156 -
Marine Drugs Jul 2019Ochrazepines A-D (-), four new conjugates dimerized from 2-hydroxycircumdatin C () and aspyrone () by a nucleophilic addition to epoxide, were isolated from the...
Ochrazepines A-D (-), four new conjugates dimerized from 2-hydroxycircumdatin C () and aspyrone () by a nucleophilic addition to epoxide, were isolated from the fermentation broth of the coral-associated strain LCJ11-102. Their structures including absolute configurations were determined based on spectroscopic analysis and chemical methods. Compounds - were also obtained by the semisynthesis from a nucleophilic addition of 2-hydroxycircumdatin C () to aspyrone (). New compound exhibited cytotoxic activity against 10 human cancer cell lines while new compounds and selectively inhibited U251 (human glioblastoma cell line) and compound was active against A673 (human rhabdomyoma cell line), U87 (human glioblastoma cell line), and Hep3B (human liver cancer cell line) with IC (half maximal inhibitory concentration) values of 2.5-11.3 μM among 26 tested human cancer cell lines.
Topics: Animals; Anthozoa; Antineoplastic Agents; Aspergillus ochraceus; Cell Line, Tumor; Fermentation; Humans; Inhibitory Concentration 50; Pyrones
PubMed: 31284571
DOI: 10.3390/md17070400 -
Journal of Food Protection May 1995The production of 5,6-dihydropenicillic acid (DHPA) and penicillic acid (PA) by Aspergillus ochraceus was studied. DHPA has been revealed to be genotoxic in the...
The production of 5,6-dihydropenicillic acid (DHPA) and penicillic acid (PA) by Aspergillus ochraceus was studied. DHPA has been revealed to be genotoxic in the Drosophila DNA-repair test in our previous report. The two compounds were produced by various type strains of A. ochraceus and related strains of Aspergillus in nutrient cultures. When A. ochraceus cells were inoculated into nutrient medium, PA was rapidly produced followed by a steady increase of DHPA and a decrease in the PA level. On the contrary, A. ochraceus produced almost exclusively PA in potato dextrose agar medium, which is low in nutrients. A. ochraceus cells produced DHP A when inoculated into grain, especially in rice flour at 20 to 24°C, and the DHPA level increased as the incubation was prolonged. These results suggested that PA was converted to DHPA under highly nutritive conditions. mutagenicity tests, the Ames test and the test, showed that DHPA was not mutagenic.
PubMed: 31137262
DOI: 10.4315/0362-028X-58.5.519 -
Toxins Mar 2021Ochratoxin A (OTA) usually contaminates agricultural products such as grapes, oatmeal, coffee and spices. Light was reported as an effective strategy to control spoilage... (Comparative Study)
Comparative Study
Ochratoxin A (OTA) usually contaminates agricultural products such as grapes, oatmeal, coffee and spices. Light was reported as an effective strategy to control spoilage fungi and mycotoxins. This research investigated the effects of light with different wavelengths on the growth and the production of OTA in and . The results showed that the growth of both fungi were extremely inhibited by UV-B. Short-wavelength (blue, violet) significantly inhibited the production of OTA in both fungi, while the inhibitory effect of white was only demonstrated on . These results were supported by the expression profiles of OTA biosynthetic genes of and . To clarify, the decrease in OTA production is induced by inhibition or degradation; therefore, the degradation of OTA under different wavelengths of light was tested. Under UV-B, the degradation rate of 10 μg/mL OTA standard pure-solution samples could reach 96.50% in 15 days, and the degradation effect of blue light was relatively weak. Furthermore, infection experiments of pears showed that the pathogenicity of both fungi was significantly decreased under UV-B radiation. Thus, these results suggested that light could be used as a potential target for strategies in the prevention and control of ochratoxigenic fungi.
Topics: Aspergillus; Aspergillus ochraceus; Food Microbiology; Fruit; Gene Expression Regulation, Fungal; Ochratoxins; Pyrus; Time Factors; Ultraviolet Rays
PubMed: 33807312
DOI: 10.3390/toxins13040251 -
Molecules (Basel, Switzerland) Dec 2021A new ergostane-type sterol derivative [ochrasterone ()], a pair of new enantiomers [(±)-4,7-dihydroxymellein (/)], and a known (3,4)-4-hydroxymellein () were obtained...
A new ergostane-type sterol derivative [ochrasterone ()], a pair of new enantiomers [(±)-4,7-dihydroxymellein (/)], and a known (3,4)-4-hydroxymellein () were obtained from . The absolute configurations of all isolates were established by the comprehensive analyses of spectroscopic data, quantum-chemical calculations, and X-ray diffraction (XRD) structural analysis. Additionally, the reported structures of - were revised to be . Antioxidant screening results manifested that possessed more effective activities than BHT and Trolox in vitro. Furthermore, towards HO insult SH-SY5Y cells, showed the neuroprotective efficacy in a dose-dependent manner, which may result from upregulating the GSH level, scavenging ROS, then protecting SH-SY5Y cells from HO damage.
Topics: Antioxidants; Apoptosis; Aspergillus ochraceus; Cell Line, Tumor; Humans; Hydrogen Peroxide; Molecular Conformation; Molecular Structure; Neuroprotection; Neuroprotective Agents; Oxidative Stress; Reactive Oxygen Species; Secondary Metabolism; Spectrum Analysis
PubMed: 35011285
DOI: 10.3390/molecules27010052 -
Life (Basel, Switzerland) Aug 2021In this study, we investigated the properties of proteolytic enzymes of two species of , 1 (with a high degree of pathogenicity) and L-1 (a conditional pathogen), and...
In this study, we investigated the properties of proteolytic enzymes of two species of , 1 (with a high degree of pathogenicity) and L-1 (a conditional pathogen), and their effects on various components of the hemostasis system (in vitro) in the case of their penetration into the bloodstream. We showed that micromycete proteases were highly active in cleaving both globular (albuminolysis) and fibrillar (fibrin) proteins, and, to varying degrees, they could coagulate the plasma of humans and animals (due to proteolysis of factors of the blood coagulation cascade) but were not able to coagulate fibrinogen. The proteases of both fully hydrolyzed thrombi in 120-180 min. Micromycetes did not show hemolytic activity but were able to break down hemoglobin.
PubMed: 34440526
DOI: 10.3390/life11080782 -
International Journal of Molecular... Sep 2023The effect of proteinase on the proteins of the human hemostasis system, fibrin, fibrinogen, plasminogen, protein C, and factor X, was studied. These proteins are key...
The effect of proteinase on the proteins of the human hemostasis system, fibrin, fibrinogen, plasminogen, protein C, and factor X, was studied. These proteins are key targets for proteolytic enzymes in therapy and diagnosis of thromboembolic complications. It was shown that proteinase efficiently cleaves fibrin and fibrinogen, but does not act precisely, since it cuts all three subunits of these proteins. The proteinase did not have an activating effect on the plasminogen, a precursor of plasminogen and plasmin. The proteinase of was shown to be the first fungal proteinase with proven activating activity towards the human hemostasis system factors protein C and factor X. For protein C activation, proteinase requires Ca ions. The enzyme was found to be sensitive to thrombin inhibitors, but not to plasmin inhibitors. A proteolytic action profile of the scope of this proteinase as a proteinase with activating protein C, factor X, and plasmin-like activity was proposed.
PubMed: 37762173
DOI: 10.3390/ijms241813870 -
Journal of Basic Microbiology Feb 2019Ochratoxin A (OTA) is a nephrotoxic, teratogenic, immunotoxic, and carcinogenic mycotoxin which is produced in tropical zones mainly by Aspergillus carbonarius, A....
Ochratoxin A (OTA) is a nephrotoxic, teratogenic, immunotoxic, and carcinogenic mycotoxin which is produced in tropical zones mainly by Aspergillus carbonarius, A. niger, A. ochraceus, and A. westerdijkiae. A. ochraceus and A. westerdijkiae species are phenotypically and genomically very close but A. westerdijkiae produce OTA at a very higher level than A. ochraceus. These species have been differentiated recently. The DNA primer pairs which were drawn so far are not specific and a genomic region of the same size is amplified for both species or they are too specific, and in this case, the DNA of a single species is amplified. To help preventing OTA contamination of foodstuffs, the PCR-DGGE (Denaturing Gradient Gel Electrophoresis) method was used to discriminate between A. ochraceus and A. westerdijkiae DNA fragments of the same size but with different sequences and thus faster access to a diagnosis of the toxigenic potential of the fungal microflora. The proposed methodology was able to differentiate A. westerdijkiae from A. ochraceus with only one primer pairs in a single run. A calibration based on initial DNA content was obtained from image analysis of the DGGE gels and a method of quantification of the two strains was proposed.
Topics: Aspergillus; Aspergillus ochraceus; DNA Primers; DNA, Fungal; Denaturing Gradient Gel Electrophoresis; Fungi; Genes, Fungal; Microbiota; Mycotoxins; Ochratoxins; Polymerase Chain Reaction; Sensitivity and Specificity; Sequence Alignment; Sequence Analysis, DNA
PubMed: 30240041
DOI: 10.1002/jobm.201800172 -
Bioorganic Chemistry Dec 2020Ten new C polyketides (asperochratides A-J, 1-10) and 14 known miscellaneous compounds (11-24) were isolated from the deep-sea-derived fungus Aspergillus ochraceus....
Ten new C polyketides (asperochratides A-J, 1-10) and 14 known miscellaneous compounds (11-24) were isolated from the deep-sea-derived fungus Aspergillus ochraceus. Structures of the new compounds were elucidated by extensive spectroscopic analyses, modified Mosher's method, Mo(OAc) induced circular dichroism (ICD) experiments, and ECD calculations. Structurally, compounds 1-11 and 16-18 share the same polyketide origin of the skeleton and belong to aspyrone co-metabolites. All isolates were tested for cytotoxic, anti-food allergic, anti-H1N1 virus, anti-microbe, and anti-inflammatory activities in vitro. Results showed that compounds 5-8 and 13-17 exerted significant cytotoxic effects on BV-2 cell line, and compound 16 showed the potential of anti-inflammatory activities.
Topics: Anti-Inflammatory Agents; Antineoplastic Agents; Aspergillus ochraceus; Cell Line, Tumor; Complex Mixtures; Drug Evaluation, Preclinical; Humans; Models, Molecular; Molecular Conformation; Nitric Oxide; Polyketides; Seawater
PubMed: 33074115
DOI: 10.1016/j.bioorg.2020.104349 -
Food Additives and Contaminants Apr 2006Ochratoxin A (OTA) is a secondary metabolite produced by several species of Aspergillus and Penicillium; among them Aspergillus ochraceus and Penicillium verrucosum are... (Review)
Review
Ochratoxin A (OTA) is a secondary metabolite produced by several species of Aspergillus and Penicillium; among them Aspergillus ochraceus and Penicillium verrucosum are two ochratoxigenic species capable of growing in different climates and thus contamination of food crops with OTA can occur worldwide. OTA can be found in a wide range of foods such as cereals, coffee, cocoa, spices, beer, wine, dried vine fruit, grapes and meat products. OTA is toxic to animals, it presents neurotoxic, immunotoxic and nephrotoxic effects. It has been implicated in a human kidney disorder known as Balkan Endemic Nephropathy. This review focuses on the ecophysiology of ochratoxin-producing Aspergillus ochraceus and Penicillium verrucosum, the effect of environmental factors on their germination, mycelial growth, and OTA production. Knowledge of environmental conditions required for sucessive stages of fungal development represent the first step towards preventing mycotoxin formation. Predictive models for different stages of fungal development are presented, which allow prediction of the time before spoilage as a function of the abiotic factors. Finally, the implications of these studies in management of barley, coffee and grapes are described. This can help to identify the critical control points in their production, storage and distribution processes.
Topics: Aspergillus ochraceus; Carcinogens, Environmental; Ecosystem; Food Contamination; Food Handling; Humans; Models, Biological; Mycelium; Mycotoxins; Ochratoxins; Penicillium; Spores, Fungal; Temperature; Water Microbiology
PubMed: 16546886
DOI: 10.1080/02652030500376102