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AIM: Potassium (K) is a key determinant for plant development and productivity. However, more than 90% of K in the soil exists in an insoluble form. K-solubilizing microbes play an important role in the transformation of insoluble K. Thus, the objective of this study was to evaluate K-dissolving ability of Aspergillus aculeatus (F) and growth-promoting properties in perennial ryegrass. METHODS AND RESULTS: Perennial ryegrass inoculated with A. aculeatus exhibited enhanced soluble K accompanied with higher growth rate and turf quality, compared with the noninoculated regimen. In addition, A. aculeatus also played a primary role in increasing chlorophyll content and photosynthetic capacity of the plant exposed to LK+F (K-feldspar plus A. aculeatus) treatment, compared with the CK (control, no K-feldspar and A. aculeatus), F (only A. aculeatus) and LK (only K-feldspar) groups. Furthermore, the antioxidase activities (CAT and POD) were significantly increased while the oxidative damage (EL and MDA) was dramatically decreased in the LK+F group compared to the LK (K-feldspar) group. Finally, in perennial ryegrass leaves, the genes expression levels of HAK8, HAK12 and HKT18 were obviously elevated in the LK+F group, compared to the CK, F and LK groups. CONCLUSION: We concluded that A. aculeatus could solubilize K from bound form and be considered as K-solubilizing biofertilizer through supplementing K in soil. SIGNIFICANCE AND IMPACT OF THE STUDY: Aspergillusaculeatus has the potential to be used as a biofertilizer in sustainable agriculture.
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Lolium , Aspergillus/genética , Fotossíntese , PotássioRESUMO
The present study aimed to assess the efficiency of silver bio-nanoparticles (Ag-NPs) in inactivating of the Aspergillus fumigatus, A. parasiticus and A. flavus var. columnaris and A. aculeatus spores. The AgNPs were synthesized in secondary metabolic products of Penicillium pedernalens 604 EAN. The inactivation process was optimized by response surface methodology (RSM) as a function of Ag NPs volume (1-10 µL/mL); time (10-120 min); pH (5-8); initial fungal concentrations (log10) (3-6). The artificial neural network (ANN) model was used to understand the behavior of spores for the factors affecting inactivation process. The best conditions to achieved SAL 10-6 of the fungal spores were recorded with 3.46 µl/mL of AgNPs, after 120 min at pH 5 and with 6 log of initial fungal spore concentrations, at which 5.99 vs. 6.09 (SAL 10-6) log reduction was recorded in actual and predicted results respectively with coefficient of 87.00%. The ANN revealed that the timehas major contribution in the inactivation process compare to Ag NPs volume. The fungal spores were totally inactivated (SAL 10-6, 6 log reduction with 99.9999%) after 110 min of the inactivation process, 10 min more was required to insure the irreversible inactivation of the fungal spores. The absence of protease and cellulase enzymes production confirm the total inactivation of the fungal spores. FESEM analysis revealed that the AgNPs which penetrated the fungal spores leading to damage and deform the fungal spore morphology. The AFM analysis confirmed the total spore surface damage. The bands in the range of the Raman spectroscopy from 1300 to 1600 cm-1 in the inactivated spores indicate the presence of CH3, CH2 and the deformation of lipids released outside the spore cytoplasm. These finding indicate that the AgNPs has high potential as a green alternative inactivation process for the airborne fungal spores.
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Nanopartículas , Penicillium , Redes Neurais de Computação , Prata , Esporos FúngicosRESUMO
Chemical epigenetic regulation (CER) is an effective method to activate the silent pathway of fungal secondary metabolite synthesis. However, conventional methods for CER study are laborious and time-consuming. In the meantime, the overall profile of the secondary metabolites in the fungi treated by the CER reagent is not well characterized. In this study, suberohydroxamic acid (SBHA), a histone deacetylase inhibitor, was added to a culture of Aspergillus aculeatus DL1011 and a new strategy based on LC-MS/MS analysis integrated with various metabolomic tools (MetaboAnalyst, MS-DIAL, SIRIUS and GNPS) was developed to characterize the profile of induced metabolites. As a result, 13.6%, 29.5% and 27.2% of metabolites were identified as newly biosynthesized, increasing and decreasing in abundance by CER, respectively. The structures of the 18 newly induced secondary metabolites were further identified by the new strategy to demonstrate that 72.2% of them (1 novel compound and 12 known compounds) were first discovered in A. aculeatus upon SBHA treatment. The accuracy of the new approach was confirmed by purification and NMR data analysis of major newly biosynthesized secondary metabolites. The bioassay showed that the newly biosynthesized compounds, roseopurpurin analogues, showed selective activities against DPPH scavenging, cytotoxicity and SHP1 inhibition. Our research demonstrated that CER was beneficial for changing the secondary metabolic profile of fungi and was an effective means of increasing the diversity of active metabolites. Our work also supplied a metabolomic strategy to characterize the profile changes and determine the newly induced compounds in the secondary metabolites of fungi treated with the chemical epigenetic regulator.
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Epigênese Genética , Espectrometria de Massas em Tandem , Cromatografia Líquida , Aspergillus/química , Metabolismo SecundárioRESUMO
This work presents the identification and proposed biosynthetic pathway for a compound of mixed polyketide-nonribosomal peptide origin that we named acurin A. The compound was isolated from an extract of the filamentous fungus Aspergillus aculeatus, and its core structure resemble that of the mycotoxin fusarin C produced by several Fusarium species. Based on bioinformatics in combination with RT-qPCR experiments and gene-deletion analysis, we identified a biosynthetic gene cluster (BGC) in A. aculeatus responsible for the biosynthesis of acurin A. Moreover, we were able to show that a polyketide synthase (PKS) and a nonribosomal peptide synthetase (NRPS) enzyme separately encoded by this BGC are responsible for the synthesis of the PK-NRP compound, acurin A, core structure. In comparison, the production of fusarin C is reported to be facilitated by a linked PKS-NRPS hybrid enzyme. Phylogenetic analyses suggest the PKS and NRPS in A. aculeatus resulted from a recent fission of an ancestral hybrid enzyme followed by gene duplication. In addition to the PKS- and NRPS-encoding genes of acurin A, we show that six other genes are influencing the biosynthesis including a regulatory transcription factor. Altogether, we have demonstrated the involvement of eight genes in the biosynthesis of acurin A, including an in-cluster transcription factor. This study highlights the biosynthetic capacity of A. aculeatus and serves as an example of how the CRISPR/Cas9 system can be exploited for the construction of fungal strains that can be readily engineered.
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Aspergillus/genética , Vias Biossintéticas/genética , Peptídeo Sintases/genética , Policetídeo Sintases/genética , Aspergillus/crescimento & desenvolvimento , Policetídeos/química , Policetídeos/metabolismoRESUMO
Agricultural or food processing wastes cause serious environmental burden and economic losses. Solid-state fermentation using these wastes is an attractive option to valorize these wastes. However, conventional autoclaving of substrate may degrade nutrients and generate toxins. Unsterilization of the substrate will cause undesired microbial contamination. Therefore, we compared irradiation with autoclaving to treat citrus wastes as substrate for solid-state fermentation by Aspergillus aculeatus. By comparing microbial growth, enzymes tested and medium consumption, irradiated substrate had higher biomass and extracellular protein, more sugar consumption and higher enzyme production than those with autoclaved substrate. Irradiation prevented the generation of cell-inhibiting components such as 5-hydroxymethylfurfural (5-HMF) whereas preserved the flavonoids well that are often enzyme inducers. These findings suggest that irradiation of agricultural and food processing wastes as substrate has advantages over autoclaving for solid-state fermentation. SIGNIFICANCE AND IMPACT OF THE STUDY: This study proposes irradiation as an alternative to sterilize agricultural residues rich in nutrients and thermosensitive compounds, such as citrus wastes for fungal solid-state fermentation and production of enzymes.
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Aspergillus/metabolismo , Citrus/metabolismo , Fermentação/fisiologia , Esterilização/métodos , Gerenciamento de Resíduos/métodos , Agricultura , Aspergillus/enzimologia , Temperatura AltaRESUMO
Considerable evidence exists that microorganisms play a significant role in the remediation of soil contaminated with heavy metals. Aspergillus aculeatus (A. aculeatus) isolated from Cd-polluted soil has been shown to increase the tolerance of turfgrasses to Cd stress. In this study, we assessed the tolerance, biosorption capacity for Cd and surface characteristics of this fungus and investigated the effect of plant inoculation with A. aculeatus on the lipid peroxidation, antioxidant activities and photosynthetic rates in rice cultivated in Cd-contaminated soil. The results indicated that the removal efficiency of A. aculeatus was 46.8% at a Cd concentration of 10â¯mgâ¯L-1. The A. aculeatus strains had the capacity to produce indole acetic acid, siderophore, and 1-aminocyclopropane-1-carboxylate deaminase and to solubilize phosphate. The O2- accumulation and the amount of MDA in rice roots inoculated with A. aculeatus were significantly lower than those in uninoculated plants. Nevertheless, no decrease in leaf ROS accumulation and photosynthetic activity was observed between the inoculated and uninoculated plants. Inoculation with A. aculeatus contained more of the ROS-scavenging metabolite GSH, a higher GSH/GSSG ratio, and higher antioxidative enzyme (SOD, POD, and CAT) activities, possibly explaining the lower ROS concentrations observed in inoculated roots in the presence of Cd. These results suggest that application of A. aculeatus has the potential to protect crops against Cd stress.
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Antioxidantes/metabolismo , Aspergillus/efeitos dos fármacos , Cádmio/análise , Oryza/microbiologia , Fotossíntese , Aspergillus/metabolismo , Dissulfeto de Glutationa/metabolismo , Ácidos Indolacéticos , Peroxidação de Lipídeos , Malondialdeído/metabolismo , Oryza/efeitos dos fármacos , Fosfatos/metabolismo , Raízes de Plantas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Sideróforos/metabolismo , Solo/química , Microbiologia do Solo , Poluentes do Solo/análiseRESUMO
Endophytes are microbes capable of colonizing the tissues of healthy plants and subsequently establishing a harmonious relationship with their hosts. In this research, the endophytic strain Paenibacillus sp. NEB was isolated from fruits of healthy Noni (Morinda citrifolia L.). Strain NEB was identified as Paenibacillus polymyxa using MALDI-TOF Mass Spectrometry. Pathogenic fungal strain NP-1 was isolated from Noni fruits infected by smut, and was identified as Aspergillus aculeatus by polyphasic taxonomy basing on morphological identification, and ITS-5.8S rDNA and ß-tubulin gene phylogenetic analyses. Through the antagonistic test against the pathogenic strain Aspergillus aculeatus NP-1, the results showed that strain NEB had a good antagonistic activity against smut pathogen of Noni. By sequencing with Illumina HiSeq 2000, the draft genome of Paenibacillus sp. NEB was acquired, and 3 CDSs for glucanases were annotated and potentially correlated to the antagonistic activity of this strain. Using realtime-PCR method with specific primers to amplify the biocontrol gene, ß-1,3-1,4- glucanase gene (gluB), it was found in Paenibacillus polymyxa NEB. This study would provide a theoretical and microbial basis for the rationally developing and using Noni beneficial microbial inoculants against its pathogenic strain in the future.
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Aspergillus/crescimento & desenvolvimento , Endófitos/fisiologia , Interações Microbianas , Morinda/microbiologia , Paenibacillus polymyxa/fisiologia , Aspergillus/classificação , Aspergillus/genética , Aspergillus/isolamento & purificação , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Endófitos/classificação , Endófitos/genética , Endófitos/isolamento & purificação , Frutas/microbiologia , Genoma Bacteriano , Anotação de Sequência Molecular , Paenibacillus polymyxa/química , Paenibacillus polymyxa/classificação , Paenibacillus polymyxa/isolamento & purificação , Filogenia , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , RNA Ribossômico 5,8S/genética , Análise de Sequência de DNA , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Tubulina (Proteína)/genéticaRESUMO
AIMS: Aspergillus aculeatus, a pathogen of peaches, can cause soft rot and lead to economic losses in agricultural production. However, studies on the prevention of soft rot caused by A. aculeatus have rarely been reported. Tricyclazole (TCZ) is a fungicide that has been widely used in disease prevention of various crops but the inhibitory mechanism of TCZ on A. aculeatus is unknown. Our aim was to determine the effects of TCZ on A. aculeatus. METHODS AND RESULTS: In our study, TCZ inhibited the growth of fungal colonies when applied at 0·5-6 mmol l-1 and inhibited the production of melanin at 3 mmol l-1 . Conidia exposed to TCZ were less effective at causing the disease in inoculated samples, and electrical conductivity, divulgation of nucleic acids and proteins rose with increasing concentrations of TCZ. Microscopic results suggest that TCZ damages not only the cell wall but also the cell membrane. Results of qRT-PCR showed that TCZ had no significant effect on the regulation of genes coding for laccase, apoptosis and hypothetical protein; however, it significantly down-regulated genes coding for cellulase, chitinase and sterol. CONCLUSIONS: Tricyclazole can influence the pathogenic ability of A. aculeatus by damaging the cell structure of hyphae and conidia, reducing the melanin production, and altering the expression of pathogenic-related gene. SIGNIFICANCE AND IMPACT OF THE STUDY: The results explained the potential cause and mechanism TCZ produced in A. aculeatus. Our research offers scientific insights into future research interest relative to using TCZ in the treatment of soft rot caused by A. aculeatus.
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An intriguing structural feature of echinocandins is the incorporation of hydroxylated amino acids. Elucidation of the machinery and the mechanism responsible for this modification is critical to generate new echinocandin derivatives with enhanced antifungal activity. In our present study, we biochemically characterized the α-ketoglutarate/Fe2+-dependent proline hydroxylase (HtyE) from two Aspergillus species, Aspergillus pachycristatus and Aspergillus aculeatus, in the respective echinocandin B and aculeacin A biosynthetic gene clusters. Our results showed that both Ap- and Aa-HtyE converted L-proline to trans-4- and trans-3-hydroxyproline, but at different ratios. Both enzymes also effectively hydroxylated C-3 of 4R-methyl-proline, L-pipecolic acid, and D-proline. Our homology modeling and site-directed mutagenesis studies identified Leu182 of Ap-HtyE as a key residue in determining the regioselectivity of Ap-HtyE. Notably, we found that the efficiency in C-3 hydroxylation of 4R-methyl-proline has no direct correlation with the ratio of trans-4-hydroxylproline to trans-3-hydroxylproline catalyzed by HtyE. Deletion of Ap-htyE abolished A. pachycristatus anti-Candida activity and the production of echinocandin B, demonstrating that HtyE is the enzyme responsible for the hydroxylation of L-proline and 4R-methyl-proline in vivo and is essential for the anti-Candida activity of echinocandin B. Our present study thus sheds light on the biochemical basis for the selective hydroxylation of L-proline and 4R-methyl-proline and reveals a new type of biocatalyst with potential for the custom production of hydroxylated proline and pipecolic acid derivatives.
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Aspergillus/enzimologia , Equinocandinas/metabolismo , Prolil Hidroxilases/genética , Prolil Hidroxilases/metabolismo , Aspergillus/classificação , Aspergillus/genética , Hidroxilação , Família Multigênica , Especificidade da EspécieRESUMO
Mining waste such as iron ore tailing is environmentally hazardous, encouraging researchers to develop effective bioremediation technologies. Among the microbial isolates collected from iron ore tailings, Aspergillus aculeatus (strain T6) showed good leaching efficiency and produced iron-containing nanoparticles under ambient conditions. This strain can convert iron ore tailing waste into agriculturally useful nanoparticles. Fourier-transform Infrared Spectroscopy (FT-IR analysis) established the at the particles are protein coated, with energy dispersive X-ray Spectroscopy (EDX analysis) showing strong signals for iron. Transmission Electron Microscopy (TEM analysis) showed semi-quasi spherical particles having average size of 15⯱â¯5â¯nm. These biosynthesized nanoparticles when tested for their efficacy on seed emergence activity of mungbean (Vigna radiata) seeds, and enhanced plant growth at 10 and 20â¯ppm.
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Aspergillus , Ferro , Nanopartículas , Biodegradação Ambiental , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
This work aimed to evaluate the effect of enzymatic pretreatment on the color and texture of plantain (Musa ssp., group AAB) dried by airflow reversal drying. Plantain slices 1.0 cm thick were used. Pretreatment with two commercial enzymes, Pectinex Ultra SPL (Aspergillus aculeatus) and Pectinex 3XL (Aspergillus niger), was performed. Drying kinetics were determined with and without pretreatment at temperatures of 50, 65 and 80 °C using a fixed bed convective dryer. An air speed of 6 m/s, a bed height of 5 cm and either unidirectional flow or airflow reversal (every 15 min) were used for drying. Color and texture were analyzed, and consumer acceptance of the results of the best treatments was determined. Pretreatment with the enzyme A. niger and airflow reversal gave the best drying kinetics and showed the greatest reduction in drying time (59.0%) at 80 °C. The best hardness results were found at 80 °C with A. niger enzymatic pretreatment with both types of air flow. Brightness and hue angle showed that samples pretreated with enzymes and dried at 65 °C had a lighter yellow color compared to non-pretreated samples. Plantain samples enzymatically pretreated and dried at 65 and 80 °C were the most accepted by consumers. This kind of enzymatic pretreatment on plantain could allow the conservation of some physical properties and reduction of drying times relative to the current methodology.
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Cellulose is the major component of the plant cell wall and the most abundant renewable biomass on earth, and its decomposition has proven to be very useful in many commercial applications. Endo-1,4-ß-d-glucanase (EC 3.2.1.4; endoglucanase), which catalyzes the random hydrolysis of 1,4-ß-glycosidic bonds of the cellulose main chain to cleave cellulose into smaller fragments, is the key cellulolytic enzyme. An endoglucanase isolated from Aspergillus aculeatus F-50 (FI-CMCase), which is classified into the glycoside hydrolase (GH) family 12, was demonstrated to be effectively expressed in the industrial strain Pichia pastoris. Here, the crystal structure and complex structures of P. pastoris-expressed FI-CMCase were solved to high resolution. The overall structure is analyzed and compared to other GH12 members. In addition, the substrate-surrounding residues were engineered to search for variants with improved enzymatic activity. Among 14 mutants constructed, one with two-fold increase in protein expression was identified, which possesses a potential to be further developed as a commercial enzyme product.
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Aspergillus/enzimologia , Celulase/química , Glicosídeo Hidrolases/química , Sequência de Aminoácidos , Aspergilose/microbiologia , Aspergillus/química , Aspergillus/genética , Aspergillus/metabolismo , Celulase/genética , Celulase/metabolismo , Cristalografia por Raios X , Expressão Gênica , Glicosídeo Hidrolases/genética , Glicosídeo Hidrolases/metabolismo , Humanos , Modelos Moleculares , Pichia/genética , Conformação Proteica , Alinhamento de Sequência , Especificidade por SubstratoRESUMO
Two extracellular polysaccharides, designated as WPA and WPB, were isolated from the fungus Aspergillus aculeatus using Q-Sepharose fast flow and Sephacryl S-300 column chromatography. WPA composed of mannose and galactose in a molar ratio of 3.9:1.0, and WPB mainly contained mannose. The molecular weight of WPA and WPB was about 28.1 kDa and 21.0 kDa, respectively. On the basis of methylation and NMR analysis, the possible main chain of WPA was [â5)-ß-D-Galf-(1 â 2,6)-α-D-Manp(1â], and WPB was mainly [â2,6)-α-D-Manp(1â], both with [α-D-Manp(1 â 2)-α-D-Manp(1 â 2)-α-D-Manp(1â] substituted at C-2 of [â2,6)-α-D-Manp(1â]. Meanwhile, WPA displayed a stronger anti-proliferative effect than WPB on HeLa, MCF-7 and MGC-803 cells in vitro. WPA and WPB could arrest HeLa cells in G2/M phase and induce HeLa cells apoptosis. Thus, our study provides evidence that WPA and WPB may be taken as potential candidates for treating cervical carcinoma.
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Apoptose/efeitos dos fármacos , Aspergillus/química , Polissacarídeos Fúngicos , Pontos de Checagem da Fase G2 do Ciclo Celular/efeitos dos fármacos , Pontos de Checagem da Fase M do Ciclo Celular/efeitos dos fármacos , Configuração de Carboidratos , Polissacarídeos Fúngicos/química , Polissacarídeos Fúngicos/isolamento & purificação , Polissacarídeos Fúngicos/farmacologia , Células HeLa , Humanos , Células MCF-7RESUMO
Aspergillus aculeatus ß-glucosidase 1 (AaBGL1) is one of the best cellobiose hydrolytic enzymes without transglycosylation products, among ß-glucosidase from various origins, for use in cellulosic biomass conversion with Trichoderma cellulases. However, in our previous report, it was demonstrated that AaBGL1 has lower catalytic efficiency toward cellobiose, which is a major end product from cellulosic biomasses by Trichoderma reesei cellulases, than do gentiobiose and laminaribiose. Thus, we expected that there is room to enhance cellobiose hydrolytic activity of AaBGL1 by increasing catalytic efficiency (k cat/K m) up to that of gentiobiose or laminaribiose for accelerating the saccharification of cellulosic biomasses, and we performed site-saturation mutagenesis targeting nine amino acids supposed to constitute subsite +1 of AaBGL1. We successfully isolated a mutant AaBGL1 (Q201E) having 2.7 times higher k cat/K m toward cellobiose than the WT enzyme. Q201E showed higher activity toward cellotriose and cellotetraose but lower activity toward gentiobiose and laminaribiose than WT. Kinetic analysis of various Q201 mutants toward cellobiose, gentiobiose, and laminaribiose revealed that only the Q201E mutation resulted in improved k cat/K m toward cellobiose. We demonstrated that side chain length and the nondissociated form of the carboxyl group at E201 in Q201E were required for enhancing the activity toward cellooligosaccharides through supporting nucleophile attack by D280 via changing catalytic environment by pH profile of kinetic parameters and mutation analyses. Moreover, we also demonstrated that Q201E produced more effective synergy with cellulases and xylanases than WT in the saccharification of alkaline-pretreated bagasse.
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Aspergillus/enzimologia , Celulose/metabolismo , Mutagênese Sítio-Dirigida , Proteínas Mutantes/metabolismo , beta-Glucosidase/metabolismo , Aspergillus/genética , Biotransformação , Celobiose/metabolismo , Concentração de Íons de Hidrogênio , Cinética , Proteínas Mutantes/genética , Especificidade por Substrato , beta-Glucosidase/genéticaRESUMO
ClbR is a Zn(II)2Cys6 transcriptional activator that controls the expression of cellulase-related genes in response to Avicel and cellobiose in Aspergillus aculeatus. A clbR-overexpressing strain (clbR-OE) that expresses the clbR gene at levels sevenfold higher than the control strain sustainably produced xylanolytic and cellulolytic activities during 10-day cultivation of A. aculeatus, enabling synchronization of xylanolytic and cellulolytic activities at a maximum level. However, clbR overexpression did not simultaneously increase levels of all xylanolytic and cellulolytic enzymes. Peptide mass fingerprint analysis revealed markedly increased production of FIa-xylanase in clbR-OE, whereas expression of FIII-avicelase and FII-carboxymethyl cellulase was unaffected and expression of hydrocellulase was lower in clbR-OE than in the control. Northern blot analysis confirmed that these effects of clbR overexpression on enzyme production were mediated at the transcriptional level. These data suggest that ClbR participates in diverse signaling pathways to control the expression of cellulosic biomass-degrading enzymes in A. aculeatus.
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Aspergillus/enzimologia , Aspergillus/genética , Biomassa , Celulase/biossíntese , Celulose/metabolismo , Endo-1,4-beta-Xilanases/biossíntese , Fatores de Transcrição/genética , Aspergillus/citologia , Aspergillus/metabolismo , Celulase/metabolismo , Endo-1,4-beta-Xilanases/metabolismo , Expressão Gênica , Transdução de Sinais/genéticaRESUMO
Aspergillus aculeatus is a common saprophyte and ubiquitous fungus belonging to section Nigri. They produce diverse secondary metabolites which are important in biological processes and industrial applications. We present the draft genome sequences of two A. aculeatus isolated from cashew nuts from coastal Kenya.
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Mucilage is a gelatinous and sticky hydrophilic polysaccharide released from epidermal cells of seed coat after the hydration of mature seeds and is composed primarily of unbranched rhamnogalacturonan I (RG-I). In this study, we produced a recombinant endo-RG-I hydrolase from Aspergillus aculeatus (AaRhgA) in the fission yeast Schizosaccharomyces pombe and examined its substrate preference for pyridylaminated (PA) RG-I with the various degrees of polymerization (DP). Recombinant AaRhgA requires PA-RG-I with a DP of 10 or higher for its hydrolase activity. We heterologously expressed the AarhgA gene under the strong constitutive promoter, cauliflower mosaic virus 35S promoter, in Arabidopsis thaliana. In a series of biochemical analyses of each mucilage fraction released from the water-imbibed seeds of the transgenic plants, we found the enhanced deposition of the transparent mucilage layer that existed in the peripheral regions of the adherent mucilage and was not stained with ruthenium red. This study demonstrated the feasibility of manipulating the mucilage organization by heterologous expression of the endo-RG-I hydrolase.
Assuntos
Arabidopsis , Aspergillus , Pectinas , Plantas Geneticamente Modificadas , Sementes , Arabidopsis/genética , Arabidopsis/metabolismo , Aspergillus/enzimologia , Aspergillus/genética , Aspergillus/metabolismo , Pectinas/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Plantas Geneticamente Modificadas/genética , Sementes/metabolismo , Mucilagem Vegetal/metabolismo , Mucilagem Vegetal/química , Schizosaccharomyces/genética , Schizosaccharomyces/metabolismo , Schizosaccharomyces/enzimologia , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/química , Regiões Promotoras Genéticas , Caulimovirus/genética , Caulimovirus/metabolismo , Glicosídeo Hidrolases/metabolismo , Glicosídeo Hidrolases/genética , Glicosídeo Hidrolases/química , Especificidade por SubstratoRESUMO
Bioremediation of extremely high-chloride wastewater poses significant challenges due to the adverse effects of elevated salt concentrations on most microorganisms, where chloride levels can be as high as 7% (w/v). Mangrove wetlands derived fungus, Aspergillus aculeatus, emerged as a promising candidate, capable of removing approximately 40% of chloride ions in environments with concentration of 15% (w/v), representative of industrial wastewater conditions. Transcriptomics and biochemical assays conducted under increasing salt conditions revealed that elevated chloride concentrations induce the expression and activity of S-adenosyl methionine-dependent methyltransferase, which facilitates the conversion of chloride into chloromethane. This is the first report characterizing the biological mechanism behind high salt tolerance and chloride removal capacity of Aspergillus aculeatus. This salt remediation mechanism may work as a starter for developing future bioremediation strategies to treat high-chloride wastewater using fungi, offering an eco-friendly alternative to traditional physical or chemical methods.
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Introduction: Potassium and phosphorus are essential macronutrients for plant growth and development. However, most P and K exist in insoluble forms, which are difficult for plants to directly absorb and utilize, thereby resulting in growth retardation of plants under P or K deficiency stress. The Aspergillus aculeatus fungus has growth-promoting characteristics and the ability to dissolve P and K. Methods: Here, to investigate the physiological effects of A. aculeatus on bermudagrass under P or K deficiency, A. aculeatus and bermudagrass were used as experimental materials. Results and discussion: The results showed that A. aculeatus could promote tolerance to P or K deficiency stress in bermudagrass, decrease the rate of leaf death, and increase the contents of crude fat as well as crude protein. In addition, A. aculeatus significantly enhanced the chlorophyll a+b and carotenoid contents. Moreover, under P or K deficiency stress, bermudagrass inoculated with A. aculeatus showed higher N, P, and K contents than non-inoculated plants. Furthermore, exogenous A. aculeatus markedly decreased the H2O2 level and CAT and POD activities. Based on our results, A. aculeatus could effectively improve the forage quality of bermudagrass and alleviate the negative effects of P or K deficiency stress, thereby playing a positive economic role in the forage industry.
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In this study, forty-two fungi from soil were isolated and tested for their carboxymethyl cellulase (CMCase) and xylanase activities. From all isolates, the fungal isolate FR60, which was identified as Aspergillus aculeatus Iizuka, showed high activities in both CMCase and xylanase with 517 mU/mg protein and 550 mU/mg protein, respectively. The crude enzyme from A. aculeatus Iizuka FR60 could hydrolyze several agricultural residues such as corncob, and sweet sorghum leaf and stalk at comparable rates with respect to the tested commercial enzymes and with a maximum rate in rice hull hydrolysis (29 µg sugar g(-1) dry weight substrate mg(-1) enzyme hr(-1)). The highest amount of glucose was obtained from corncob by using the crude enzyme from A. aculeatus Iizuka FR60 (10.1 g/100 g dry substrate). From overall enzymatic treatment results, the lowest sugar yield was from rice hulls treatment (1.6 g/100 g dry weight) and the highest amount of reducing sugar was obtained from rice straw treatment (15.3 g/100 g dry weight). Among tested agricultural wastes, rice hull could not be effectively hydrolyzed by enzymes, whereas sugarcane leaf and stalk, and peanut shell could be effectively hydrolyzed (30-31% total sugar comparing with total sugar yield from acid treatment).