RESUMEN
Fungal pathogens pose significant threats to plant health by secreting effectors that manipulate plant-host defences. However, identifying effector proteins remains challenging, in part because they lack common sequence motifs. Here, we introduce Fungtion (Fungal effector prediction), a toolkit leveraging a hybrid framework to accurately predict and visualize fungal effectors. By combining global patterns learned from pretrained protein language models with refined information from known effectors, Fungtion achieves state-of-the-art prediction performance. Additionally, the interactive visualizations we have developed enable researchers to explore both sequence- and high-level relationships between the predicted and known effectors, facilitating effector function discovery, annotation, and hypothesis formulation regarding plant-pathogen interactions. We anticipate Fungtion to be a valuable resource for biologists seeking deeper insights into fungal effector functions and for computational biologists aiming to develop future methodologies for fungal effector prediction: https://step3.erc.monash.edu/Fungtion/.
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Biología Computacional , Proteínas Fúngicas , Proteínas Fúngicas/metabolismo , Proteínas Fúngicas/química , Biología Computacional/métodos , Programas Informáticos , Hongos/metabolismo , Hongos/química , Interacciones Huésped-Patógeno , Plantas/microbiología , Plantas/metabolismoRESUMEN
Extracellular vesicles (EVs) are vesicle-like structures composed of lipid bilayers, which can be divided into apoptotic bodies, microbubbles and exosomes. They are nanoparticles used for the exchange of information between cells. EVs contains many substances, including protein. With the development of proteomics, we know more about the types and functions of protein in vesicles. The potential functions of proteins in the envelope are mainly discussed, including cell wall construction, fungal virulence transmission, signal transmission and redox reactions, which provides a new perspective for studying the interaction mechanism between fungi and other organisms. The fungal protein markers of EVs are also summarized, which provided an exploration tool for studying the mechanism of vesicles. In addition, the possible role of immune protein in the EVs in the treatment of human diseases is also discussed, which provides new ideas for vaccine development.
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Vesículas Extracelulares , Proteínas Fúngicas , Hongos , Vesículas Extracelulares/metabolismo , Vesículas Extracelulares/química , Proteínas Fúngicas/metabolismo , Proteínas Fúngicas/química , Humanos , Hongos/metabolismo , Hongos/química , Proteómica/métodosRESUMEN
Nature uses compact but functionalized biosynthetic fragments as building blocks to generate complex natural products. To leverage this strategy for the discovery of natural products with new scaffolds, we performed genome mining to identify biosynthetic gene clusters (BGCs) in fungi that embed genes that can synthesize targeted fragments. The three-enzyme pathway that biosynthesizes the strained dityrosine cyclophane in the herquline A pathway was used to identify a large number of potential BGCs that may use the cyclophane as a fragment. Characterization of a conserved BGC from fungal strains led to the isolation of octacyclin A, an octacyclic natural product with an unprecedented structure, including two hetero-[3.3.1]bicycles and a combination of fused, bridged, and macrocyclic rings. Biosynthetic steps leading to octacyclin A were fully elucidated using pathway reconstitution and enzymatic assays, unveiling intriguing chemical logic and new enzymatic reactions in building the octacyclic core. Our work demonstrates the potential utility of fragment-guided genome mining in expanding natural product chemical space.
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Alcaloides , Familia de Multigenes , Alcaloides/química , Hongos/genética , Hongos/química , Genoma Fúngico , Productos Biológicos/química , Productos Biológicos/metabolismo , CiclofanosRESUMEN
Elemental profiling of fungal species as a phenotyping tool is an understudied topic and is typically performed to examine plant tissue or non-biological materials. Traditional analytical techniques such as inductively coupled plasma-optical emission spectroscopy (ICP-OES) and inductively coupled plasma-mass spectrometry (ICP-MS) have been used to identify elemental profiles of fungi; however, these techniques can be cumbersome due to the difficulty of preparing samples. Additionally, the instruments used for these techniques can be expensive to procure and operate. Laser-induced breakdown spectroscopy (LIBS) is an alternative elemental analytical technique-one that is sensitive across the periodic table, easy to use on various sample types, and is cost-effective in both procurement and operation. LIBS has not been used on axenic filamentous fungal isolates grown in substrate media. In this work, as a proof of concept, we used LIBS on two genetically distinct fungal species grown on a nutrient-rich and nutrient-poor substrate media to determine whether robust elemental profiles can be detected and whether differences between the fungal isolates can be identified. Our results demonstrate a distinct correlation between fungal species and their elemental profile, regardless of the substrate media, as the same strains shared a similar uptake of carbon, zinc, phosphorus, manganese, and magnesium, which could play a vital role in their survival and propagation. Independently, each fungal species exhibited a unique elemental profile. This work demonstrates a unique and valuable approach to rapidly phenotype fungi through optical spectroscopy, and this approach can be critical in understanding these fungi's behavior and interactions with the environment. IMPORTANCE: Historically, ionomics, the elemental profiling of an organism or materials, has been used to understand the elemental composition in waste materials to identify and recycle heavy metals or rare earth elements, identify the soil composition in space exploration on the moon or Mars, or understand human disorders or disease. To our knowledge, ionomic profiling of microbes, particularly fungi, has not been investigated to answer applied and fundamental biological questions. The reason is that current ionomic analytical techniques can be laborious in sample preparation, fail to measure all potential elements accurately, are cost-prohibitive, or provide inconsistent results across replications. In our previous efforts, we explored whether laser-induced breakdown spectroscopy (LIBS) could be used in determining the elemental profiles of poplar tissue, which was successful. In this proof-of-concept endeavor, we undertook a transdisciplinary effort between applied and fundamental mycology and elemental analytical techniques to address the biological question of how LIBS can used for fungi grown axenically in a nutrient-rich and nutrient-poor environment.
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Hongos , Rayos Láser , Análisis Espectral , Análisis Espectral/métodos , Análisis Espectral/instrumentación , Hongos/aislamiento & purificación , Hongos/química , Hongos/clasificación , Hongos/metabolismo , Espectrometría de Masas/métodosRESUMEN
Medicinal plants are "goldmines" of natural products, and continue to provide key scaffolds for drug development. They have immense therapeutic potential, encapsulating a plethora of metabolites within them, which have yet to be explored. Withania somnifera (L.) Dunal is one such medicinal plant known since time immemorial for its therapeutic activity in the Ayurveda system of medicine. Studies have revealed Nature's marvel of these medicinal plants harbouring endophytic and epiphytic microorganisms from phyllosphere to rhizosphere. Chromatographic fingerprinting was carried out using HPTLC and HPLC on five epiphytic strains isolated from the leaves, stem and fruits of Withania somnifera. Out of five filamentous fungi, one fungus identified as Aspergillus aculeatus S20 was well explored. An indole alkaloid, okaramine H, was isolated using systematic chromatographic investigation at a retention time of 26.278 min showing λmax at 206, 236, 284 and 370 nm. Confirmation was achieved using NMR and mass spectrometry (MS) as analytical techniques. Structure elucidation was done by studying the fragmentation pattern using MS/MS and an accurate mass was determined using HR-ESI-QTOF-MS showing m/z of 521.2546 [M + H]+. The percentage purity of isolated okaramine H was found to be >90. Well known for its insecticidal activity, okaramine H was explored for its antileishmanial activity against the Leishmania donovani parasite for the first time. Under in vitro conditions, the compound showed an inhibitory effect on Leishmania donovani promastigotes with an IC50 of 147 µg mL-1.
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Withania , Withania/química , Cromatografía Líquida de Alta Presión/métodos , Cromatografía en Capa Delgada/métodos , Alcaloides Indólicos/farmacología , Alcaloides Indólicos/química , Alcaloides Indólicos/aislamiento & purificación , Aspergillus/química , Aspergillus/efectos de los fármacos , Aspergillus/aislamiento & purificación , Animales , Hojas de la Planta/microbiología , Hojas de la Planta/química , Hongos/química , Hongos/efectos de los fármacos , Hongos/aislamiento & purificación , Leishmania donovani/efectos de los fármacos , Espectrometría de Masas en Tándem/métodosRESUMEN
OBJECTIVES: Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is extensively employed for the identification of filamentous fungi on MALDI Biotyper (Bruker Daltonics) and Vitek MS (biomerieux), but the performance of fungi identification on new EXS2600 (Zybio) is still unknow. Our study aims to evaluate the new EXS2600 system's (Zybio) ability to rapidly identify filamentous fungi and determine its effect on turnaround time (TAT) in our laboratory. METHODS: We tested 117 filamentous fungi using two pretreatment methods: the formic acid sandwich (FA-sandwich) and a commercial mold extraction kit (MEK, Zybio). All isolates were confirmed via sequence analysis. Laboratory data were extracted from our laboratory information system over two 9-month periods: pre-EXS (April to December 2022) and post-EXS (April to December 2023), respectively. RESULTS: The total correct identification (at the species, genus, or complex/group level) rate of fungi was high, FA-sandwich (95.73%, 112/117), followed by MEK (94.02%, 110/117). Excluding 6 isolates not in the database, species-level identification accuracy was 92.79% (103/111) for FA-sandwich and 91.89% (102/111) for MEK; genus-level accuracy was 97.29% (108/111) and 96.39% (107/111), respectively. Both methods attained a 100% correct identification rate for Aspergillus, Lichtheimia, Rhizopus Mucor and Talaromyces species, and were able to differentiate between Fusarium verticillioides and Fusarium proliferatum within the Fusarium fujikuroi species complex. Notably, high confidence was observed in the species-level identification of uncommon fungi such as Trichothecium roseum and Geotrichum candidum. The TAT for all positive cultures decreased from pre EXS2600 to post (108.379 VS 102.438, P < 0.05), and the TAT for tissue decreased most (451.538 VS 222.304, P < 0.001). CONCLUSIONS: The FA-sandwich method is more efficient and accurate for identifying filamentous fungi with EXS2600 than the MEK. Our study firstly evaluated the performance of fungi identification on EXS2600 and showed it is suitable for clinical microbiology laboratories use.
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Formiatos , Hongos , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Hongos/clasificación , Hongos/aislamiento & purificación , Hongos/química , Hongos/genética , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Formiatos/químicaRESUMEN
Fu Brick Tea (FBT) is characterized by Fungus Aroma (FA), which determines the quality of FBT products. However, the aroma constituents and their interactive mechanism for FA remain unclear. In this study, the FBT sample with the optimal FA characteristics was selected from 29 FBTs. Then, 19 components with OAV ≥ 1 were identified as the odorants involved in the FA formation. The aroma recombination test suggested that the FA was potentially produced by the synergistic interplay among the 15 key odorants, including (E,E)-2,4-heptadienal, (E,E)-2,4-nonadienal, (E)-2-nonenal, (E,Z)-2,6-nonadienal, (E)-2-octenal, (E)-ß-ionone, 4-ketoisophorone, dihydroactinidiolide, (E)-ß-damascenone, 1-octen-3-ol, linalool, geraniol, heptanal, hexanal, and phenylacetaldehyde. And, the synergistic effects between them were preliminarily studied by aroma omissions, such as modulatory effects, masking effects, compensatory effects, and novelty effects, ultimately contributing to the FA. In all, this work helps us better understand the formation of the FA and provides a basis for the improvement of FBT production technology.
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Camellia sinensis , Odorantes , Té , Compuestos Orgánicos Volátiles , Odorantes/análisis , Té/química , Té/microbiología , Compuestos Orgánicos Volátiles/química , Compuestos Orgánicos Volátiles/metabolismo , Camellia sinensis/química , Camellia sinensis/microbiología , Hongos/metabolismo , Hongos/química , Cromatografía de Gases y Espectrometría de Masas , Humanos , Aromatizantes/química , Aromatizantes/metabolismoRESUMEN
Natural bioactive compounds encompass a vast array of molecules derived from plants, fungi, marine organisms, and other natural sources [...].
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Productos Biológicos , Productos Biológicos/química , Productos Biológicos/farmacología , Humanos , Plantas/química , Organismos Acuáticos/química , Hongos/químicaRESUMEN
Cellular agriculture, an alternative and innovative approach to sustainable food production, has gained momentum in recent years. However, there is limited research into the production of cultivated seafood. Here, we investigated the ability of fish mackerel cells (Scomber scombrus) to adhere to plant, algal and fungal-based biomaterial scaffolds, aiming to optimize the cultivation of fish cells for use in cellular agriculture. A mackerel cell line was utilized, and metabolic assays and confocal imaging were utilized to track cell adhesion, growth, and differentiation on the different biomaterials. The mackerel cells adhered and grew on gelatin (positive control), zein, and soy proteins, as well as on alginate, chitosan, and cellulose polysaccharides. The highest adhesion and growth were on the zein and chitosan substrates, apart from the gelatin control. These findings provide a blueprint to enhance scaffold selection and design, contributing to the broader field of cellular agriculture through the development of scalable and eco-conscious solutions for meeting the growing global demand for seafood.
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Materiales Biocompatibles , Adhesión Celular , Animales , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Adhesión Celular/efectos de los fármacos , Gelatina/química , Quitosano/química , Hongos/metabolismo , Hongos/química , Plantas/química , Plantas/metabolismo , Plantas/microbiología , Línea Celular , Alginatos/química , Peces , Andamios del Tejido/químicaRESUMEN
Saccharide mapping was a promising scheme to unveil the mystery of polysaccharide structure by analysis of the fragments generated from polysaccharide decomposition process. However, saccharide mapping was not widely applied in the polysaccharide analysis for lacking of systematic introduction. In this review, a detailed description of the establishment process of saccharide mapping, the pros and cons of downstream technologies, an overview of the application of saccharide mapping, and practical strategies were summarized. With the updating of the available downstream technologies, saccharide mapping had been expanding its scope of application to various kinds of polysaccharides. The process of saccharide mapping analysis included polysaccharides degradation and hydrolysates analysis, and the degradation process was no longer limited to acid hydrolysis. Some downstream technologies were convenient for rapid qualitative analysis, while others could achieve quantitative analysis. For the more detailed structure information could be provided by saccharide mapping, it was possible to improve the quality control of polysaccharides during preparation and application. This review filled the blank of basic information about saccharide mapping and was helpful for the establishment of a professional workflow for the saccharide mapping application to promote the deep study of polysaccharide structure.
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Polisacáridos Fúngicos , Plantas , Polisacáridos Fúngicos/química , Hidrólisis , Plantas/química , Polisacáridos/química , Hongos/químicaRESUMEN
Mycoplasma genitalium is a sexually transmitted bacterium associated with urogenital disease syndromes in the US and worldwide. The global rise in drug resistance in M. genitalium necessitates the development of novel drugs to treat this pathogen. To address this need, we have screened extracts from a library of fungal isolates assembled through the University of Oklahoma Citizen Science Soil Collection Program. Analysis of one of the bioactive extracts using bioassay-guided fractionation led to the purification of the compound PF1140 (1) along with a new and several other known pyridones. The N-hydroxy pyridones are generally regarded as siderophores with high binding affinity for iron(III) under physiological conditions. Results from UV-vis absorption spectroscopy-based titration experiments revealed that 1 complexes with Fe3+. As M. genitalium does not utilize iron, we propose that the PF1140-iron complex induces cytotoxicity by facilitating the cellular uptake of iron, which reacts with endogenous hydrogen peroxide to produce toxic hydroxyl radicals.
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Hierro , Mycoplasma genitalium , Mycoplasma genitalium/metabolismo , Mycoplasma genitalium/efectos de los fármacos , Hierro/metabolismo , Estructura Molecular , Pruebas de Sensibilidad Microbiana , Antibacterianos/farmacología , Antibacterianos/química , Piridonas/farmacología , Piridonas/química , Hongos/química , Hongos/metabolismo , Compuestos Férricos/química , Peróxido de Hidrógeno , Sideróforos/farmacología , Sideróforos/químicaRESUMEN
Chitin is a polymer of N-acetylglucosamine and an essential component of the fungal cell wall. Chitosan is the deacetylated form of chitin and is also important for maintaining the integrity of this structure. Both polysaccharides are widely distributed in nature and have been shown to have a variety of applications in biomedicine, including their potential in immune sensing and as potential antifungal agents. In addition, chitin has been reported to play an important role in the pathogen-host interaction, involving innate and adaptive immune responses. This paper will explore the role of chitin and chitosan when incorporated into nanobiocomposites to improve their efficacy in detecting fungi of medical interest and inhibiting their growth. Potential applications in diagnostic and therapeutic medicine will be discussed, highlighting their promise in the development of more sensitive and effective tools for the early diagnosis of fungal infections. This review aims to highlight the importance of the convergence of nanotechnology and biology in addressing public health challenges.
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Antifúngicos , Quitina , Quitosano , Hongos , Quitina/química , Quitina/farmacología , Quitosano/química , Quitosano/farmacología , Antifúngicos/farmacología , Antifúngicos/química , Hongos/efectos de los fármacos , Hongos/química , Humanos , Nanocompuestos/química , Micosis/inmunología , Micosis/tratamiento farmacológico , Micosis/diagnósticoRESUMEN
Fungi pose a global threat to humankind due to the increasing emergence of multi-drug-resistant fungi. There is a rising incidence of invasive fungal infections. Due to the structural complexity of fungal cell membranes, only a few classes of antifungal agents are effective and have been approved by the U.S. FDA. Hence, researchers globally are focusing on developing novel strategies to cure fungal infections. One of the potential strategies is the "Trojan horse" approach, which uses the siderophore-mediated iron acquisition (SIA) system to scavenge iron to deliver potent antifungal agents for therapeutics and diagnostics. These siderophore conjugates chelate to iron and are taken up through siderophore-iron transporters, which are overexpressed exclusively on microbes such as bacteria or fungi, but not mammalian cells. Our comprehensive review delves into recent advancements in the design of siderophore-conjugated antifungal agents to gain fungal cell entry. Notably, our focus extends to unraveling the intricate relationship between the structure of natural siderophores or siderophore-like molecules and the resulting antifungal activity. By exploring these design strategies, we aim to contribute to the ongoing discourse on combating drug-resistant fungal infections and advancing the landscape of antifungal theranostics.
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Antifúngicos , Hongos , Micosis , Sideróforos , Sideróforos/química , Sideróforos/farmacología , Antifúngicos/farmacología , Antifúngicos/química , Humanos , Micosis/tratamiento farmacológico , Micosis/microbiología , Hongos/efectos de los fármacos , Hongos/química , Hierro/metabolismo , Hierro/química , AnimalesRESUMEN
Covering: up to December 2023Decalin-containing tetramic acid derivatives, especially 3-decalinoyltetramic acids (3-DTAs), are commonly found as fungal secondary metabolites. Numerous biological activities of this class of compounds, such as antibiotic, antiviral, antifungal, antiplasmodial, and antiprotozoal properties, have been the subject of ongoing research. For this reason, these molecules have attracted a lot of interest from the scientific community and various efforts including semi-synthesis, co-culturing with bacteria and biosynthetic gene sequencing have been made to obtain more derivatives. In this review, 3-DTAs are classified into four major groups based on the absolute configuration of the bicyclic decalin ring. Their biosynthetic pathways, various biological activities, and structure-activity relationship are then introduced.
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Hongos , Pirrolidinonas , Relación Estructura-Actividad , Hongos/química , Hongos/metabolismo , Pirrolidinonas/farmacología , Pirrolidinonas/química , Pirrolidinonas/aislamiento & purificación , Pirrolidinonas/metabolismo , Estructura Molecular , Naftalenos/farmacología , Naftalenos/química , Naftalenos/aislamiento & purificación , Naftalenos/metabolismo , Antifúngicos/farmacología , Antifúngicos/química , Antifúngicos/aislamiento & purificación , Antibacterianos/farmacología , Antibacterianos/química , Antibacterianos/biosíntesis , Antibacterianos/aislamiento & purificaciónRESUMEN
A magnetic biochar nanomaterial derived from fungal hyphae was introduced into the sample preparation field. The magnetic fungal hyphae-derived biomass carbon (MFHBC) could be produced by a controllable hydrothermal method. In order to obtain the best sorbent for magnetic solid-phase extraction (MSPE), the reaction conditions containing temperature, time and the consumption of fungal hyphae were investigated. A series of MFHBC materials were characterized by vibrating sample magnetometers, X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy, and transmission electron microscopy. A material with a satisfactory saturation magnetization (21.58 emu g-1) and largest surface area (88.06 m2 g-1) was selected as the sorbent to extract ten typical organochlorine pesticides (OCPs). The extraction conditions were optimized as 20 mL of sample solution with 70 mg of sorbent and 2.0 g of NaCl oscillated at 50 °C for 5.0 min. And the optimum desorption was performed by oscillating sorbent in 1.0 mL acetonitrile for 5.0 min. Then, the MFHBC-based MSPE-GC-MS/MS methods were established for different samples including water samples, tea beverages, and Chinese traditional medicines. The linearities were 10-2500 ng L-1 or 100-25,000 ng kg-1, and the limits of detection were 0.3-13.9 ng L-1 for water sample, 0.1-9.7 ng L-1 for tea beverage samples, 0.1-21.4 ng L-1 for Shenqi Fuzheng injection samples, and 7.2-278.3 ng kg-1 for Astragali Radix decoction pieces. Except for satisfactory repeatability (RSDs ≤13.8%) in intra-day and inter-day tests (n = 3), the reproducibility (RSDs ≤13.5%, n = 3) of MFHBC was acceptable. The methods were applied in the determination of OCPs from above real samples, with the recoveries of 80.5-117.2% and the RSDs (n = 3) <8.9%. The methods were suitable in the sensitive determination of OCPs from simple to complex matrix samples.
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Hidrocarburos Clorados , Extracción en Fase Sólida , Espectrometría de Masas en Tándem , Extracción en Fase Sólida/métodos , Hidrocarburos Clorados/química , Hidrocarburos Clorados/análisis , Hidrocarburos Clorados/aislamiento & purificación , Té/química , Plaguicidas/química , Cromatografía de Gases y Espectrometría de Masas , Contaminantes Químicos del Agua/química , Contaminantes Químicos del Agua/aislamiento & purificación , Carbono/química , Carbón Orgánico/química , Medicina Tradicional China , Hongos/química , Hongos/aislamiento & purificación , Medicamentos Herbarios Chinos/química , Biomasa , Contaminación de Alimentos/análisis , Residuos de Plaguicidas/químicaRESUMEN
Inflammation and associated disorders have been a major contributing factor to mortality worldwide. The augmented mortality rate and emerging resistance against the approved therapeutics necessitate the discovery of novel chemistries destined for multiple clinical settings. Cellular factories including endophytic fungi have been tapped for chemical diversity with therapeutic potential. The emerging evidence has suggested the potential of bioactive compounds isolated from the endophytic fungi as putative agents to combat inflammation-associated disorders. The review summarizesand assists the readers in comprehending the structural and functional aspects of the medicinal chemistries identified from endophytic fungi as anticancer, antiobesity, antigout, and immunomodulatory agents.
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Hongos , Humanos , Hongos/química , Antineoplásicos/farmacología , Antineoplásicos/química , Animales , Endófitos/química , Endófitos/metabolismo , Estructura Molecular , Fármacos Antiobesidad/farmacología , Fármacos Antiobesidad/química , Fármacos Antiobesidad/aislamiento & purificación , Productos Biológicos/química , Productos Biológicos/farmacología , Productos Biológicos/aislamiento & purificación , Factores Inmunológicos/farmacología , Factores Inmunológicos/químicaRESUMEN
Glycogen is a highly branched glucose polymer that is an energy storage material in fungi and animals. Extraction of glycogen from its source in a way that minimizes its molecular degradation is essential to investigate its native structure. In this study, the following extraction methods were compared: sucrose gradient density ultracentrifugation, thermal alkali, hot alcohol and hot water extractions. Molecular-size and chain-length distributions of glycogen were measured by size-exclusion chromatography and fluorophore-assisted carbohydrate electrophoresis, respectively. These two fine-structure features are the most likely structural characteristics to be degraded during extraction. The results show that the thermal alkali, hot alcohol and hot water extractions degrade glycogen molecular size and/or chain-length distributions, and that sucrose gradient density ultracentrifugation with neither high temperature nor alkaline treatment is the most suitable method for fungal glycogen extraction.
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Glucógeno , Glucógeno/química , Glucógeno/metabolismo , Hongos/química , Peso Molecular , Fraccionamiento Químico/métodos , Cromatografía en Gel/métodos , Ultracentrifugación/métodosRESUMEN
Extracellular vesicles (EVs) have been identified in diverse fungi, including human pathogens. In this protocol, we present two techniques for isolating and analyzing fungal EVs. The first is for high-throughput screening, and the second is for yielding concentrated samples suitable for centrifugation-based density gradients. We describe steps for analytical assays such as nano-flow cytometry and nanoparticle tracking analysis to measure EV dimensions and concentration. EV suspensions can serve diverse assays, including electron microscopy, compositional determination, and cell-to-cell communication assays. For complete details on the use and execution of this protocol, please refer to Rizzo et al.,1 Rizzo et al.,2 Reis et al.,3 and Reis et al.4.
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Vesículas Extracelulares , Hongos , Ultracentrifugación , Vesículas Extracelulares/química , Vesículas Extracelulares/metabolismo , Ultracentrifugación/métodos , Hongos/química , Hongos/metabolismo , Hongos/aislamiento & purificación , Hongos/citología , Citometría de Flujo/métodos , Medios de Cultivo/químicaRESUMEN
Spices are food flavouring agents that are highly used in Iraq. However, they may be contaminated by toxicogenic fungi and subsequent production of mycotoxins. The aim of this study was to investigate the contamination of commonly used spices with fungi using polymerase chain reaction (PCR) assay and to detect fungal mycotoxin using high-performance liquid chromatography. Thirty-five spices (seven samples from each black pepper, red pepper, turmeric, cumin and ginger) were cultured on an appropriate medium to identify various fungi species. Later on, the toxigenicity of Aspergillus flavus and Aspergillus niger was determined using a PCR assay. The fungal mycotoxins, including aflatoxins and ochratoxins, were then determined through a high-performance liquid chromatography using the validated Quick, Easy, Cheap, Effective, Rugged, and Safe (QuECHERS) method. Aspergillus species were the predominantly isolated fungi, followed by Penicillium and Fusarium. The PCR results indicate the high toxigenicity of A. flavus as 85.7% of the strains had aflQ/aflR genes and 79% had PKS15KS/PKS15C-MeT genes. Regarding mycotoxin contamination in spices, the highest rates of aflatoxins and ochratoxins were found in black pepper (5.913 µg/kg) and red chilli (6.9055 µg/kg), respectively. Spices are susceptible substrates for the growth of mycotoxigenic fungi. Thus, regular effective surveillance and quality control procedures are highly recommended.
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Hongos , Micotoxinas , Especias , Especias/análisis , Especias/microbiología , Micotoxinas/análisis , Cromatografía Líquida de Alta Presión/métodos , Hongos/genética , Hongos/aislamiento & purificación , Hongos/química , Hongos/clasificación , Hongos/metabolismo , Reproducibilidad de los Resultados , Límite de Detección , Reacción en Cadena de la Polimerasa/métodos , Modelos LinealesRESUMEN
Three new indole alkaloids, named talatensindoids A-C (1-3), together with two known biogenetically related indole alkaloids tryptamine (4) and L-tryptophan (5) were isolated from the Talaromyces assiutensis JTY2 based on the guidance of OSMAC approach. The structures of these indole alkaloids were determined by comprehensive spectroscopic analyses. The absolute configuration of 3 was confirmed by X-ray crystallographic analysis. Compound 1 represent the rare example of a chlorine-substituted indole alkaloid from natural products. The inhibitory activity of compounds 1-5 against two phytopathogenic fungi and three phytopathogenic bacteria was evaluated. Compound 1 exhibited broad spectrum antibacterial activities.