scholarly journals Characterization of Hydroxyproline-Containing Hairpin-Like Antimicrobial Peptide EcAMP1-Hyp from Barnyard Grass (Echinochloa crusgalli L.) Seeds: Structural Identification and Comparative Analysis of Antifungal Activity

2018 ◽  
Vol 19 (11) ◽  
pp. 3449 ◽  
Author(s):  
Eugene Rogozhin ◽  
Artur Zalevsky ◽  
Alexander Mikov ◽  
Alexey Smirnov ◽  
Tsezi Egorov
Keyword(s):  
Antifungal Activity ◽  
Plant Immunity ◽  
Pathogenic Fungus ◽  
Hyphal Growth ◽  
Single Amino Acid ◽  
Hydrophobic Core ◽  
Barnyard Grass ◽  
Vitro Binding ◽  
Dynamics Simulations

Herein, we describe a modified form of the antimicrobial hairpin-like peptide EcAMP1, isolated from barnyard grass (E. crusgalli) seeds, which is structurally characterized by a combination of high-pressure liquid chromatography, mass spectrometry, and automated Edman sequencing. This derivate has a single amino acid substitution (Pro19Hyp) in the second α-helical region of the molecule, which is critical for the formation of the hydrophobic core and the secondary structure elements. Comparing the antifungal activity of these two peptides, we found that the modified EcAMP1-Hyp had a significantly weaker activity towards the most-sensitive plant pathogenic fungus Fusarium solani. Molecular dynamics simulations and in vitro binding to the commercial polysaccharides allowed us to conclude that the Pro-19 residue is important for binding to carbohydrates located in the spore cell wall and it chiefly exhibits a fungistatic action representing the hyphal growth inhibition. These data are novel and significant for understanding a role of α-hairpinins in plant immunity.

scholarly journals Mass spectrometric quantification of the adaptations in the wall proteome of Candida albicans in response to ambient pH

Microbiology ◽  
2011 ◽  
Vol 157 (1) ◽  
pp. 136-146 ◽  
Author(s):  
Grazyna J. Sosinska ◽  
Leo J. de Koning ◽  
Piet W. J. de Groot ◽  
Erik M. M. Manders ◽  
Henk L. Dekker ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Candida Albicans ◽  
Pathogenic Fungus ◽  
Hyphal Growth ◽  
Mass Spectrometric ◽  
Spectrometric Analysis ◽  
Mucosal Surfaces ◽  
Covalently Linked ◽  
Semi Solid

The mucosal layers colonized by the pathogenic fungus Candida albicans differ widely in ambient pH. Because the properties and functions of wall proteins are probably pH dependent, we hypothesized that C. albicans adapts its wall proteome to the external pH. We developed an in vitro system that mimics colonization of mucosal surfaces by growing biomats at pH 7 and 4 on semi-solid agarose containing mucin as the sole nitrogen source. The biomats expanded radially for at least 8 days at a rate of ∼30 μm h−1. At pH 7, hyphal growth predominated and growth was invasive, whereas at pH 4 only yeast and pseudohyphal cells were present and growth was noninvasive. Both qualitative mass spectrometric analysis of the wall proteome by tandem mass spectrometry and relative quantification of individual wall proteins (pH 7/pH 4), using Fourier transform mass spectrometry (FT-MS) and a reference mixture of 15N-labelled yeast and hyphal walls, identified similar sets of >20 covalently linked wall proteins. The adhesion proteins Als1 and Als3, Hyr1, the transglucosidase Phr1, the detoxification enzyme Sod5 and the mammalian transglutaminase substrate Hwp1 (immunological detection) were only present at pH 7, whereas at pH 4 the level of the transglucosidase Phr2 was >35-fold higher than at pH 7. Sixteen out of the 22 proteins identified by FT-MS showed a greater than twofold change. These results demonstrate that ambient pH strongly affects the wall proteome of C. albicans, show that our quantitative approach can give detailed insights into the dynamics of the wall proteome, and point to potential vaccine targets.

scholarly journals Antifungal activity of Bacillussp. Gn-A11-18isolated from decomposing solid green household waste in water and soil against Candida albicans and Aspergillus niger

2020 ◽  
Vol 150 ◽  
pp. 02003 ◽  
Author(s):  
Azeddin El Barnossi ◽  
Fatimazahrae Moussaid ◽  
Abdelilah Iraqi Housseini
Keyword(s):  
Candida Albicans ◽  
Antifungal Activity ◽  
Aspergillus Niger ◽  
Pathogenic Fungus ◽  
Bioactive Compound ◽  
Household Waste ◽  
Diffusion Method ◽  
Agar Plug ◽  
Opportunistic Pathogenic

Candida albicans an opportunistic pathogenic fungus causes many infections in humans. Whereas Aspergillus niger is a fungus that can produce ochratoxins, a group of extremely dangerous secondary metabolites that are classified as potentially carcinogenic to humans and also causing deterioration in grapes, strawberries, etc. The purpose of the current study is to isolate, purify, identify and characterize new microorganisms associated with solid green household waste for the control of C. albicans and A. niger. The antifungal activity of bacterial isolates was carried out in vitro by the agar plug diffusion method, the disk and well diffusion method. The isolate that showed promising activity has been identified by those macroscopic, microscopic and biochemical characteristics. The results obtained in the course of this study showed the isolation of an isolate named Gn-A11-18, which was shown to have significant inhibitory activity with a 42.66% inhibition percentage against A. niger and a 44.66 mm inhibition diameter against C. albicans compared to the controls. The identification of Gn-A11-18 isolate has shown that this isolate belongs to the genus Bacillus with a similarity to Bacillus subtilis and Bacillus tequilensis. In the light of the results of this study, we can suggest that the bioactive compound of Bacillus sp. Gn-A11-18 could become a biological alternate that could have an important role to fight against C. albicans and A. niger.

In vitro and in vivo activity of chelerythrine against Candida albicans and underlying mechanisms

2019 ◽  
Vol 14 (18) ◽  
pp. 1545-1557 ◽  
Author(s):  
Ying Gong ◽  
Siwen Li ◽  
Weixin Wang ◽  
Yiman Li ◽  
Wenli Ma ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Antifungal Activity ◽  
Calcium Concentration ◽  
Galleria Mellonella ◽  
Hyphal Growth ◽  
Drug Transporter ◽  
Antifungal Effect ◽  
Underlying Mechanisms

Aim: To evaluate whether chelerythrine (CHT) exhibited antifungal activity against Candida albicans in vitro and in vivo and to explore the underlying mechanisms. Materials & methods: Broth microdilution assay and Galleria mellonella model were used to evaluate the antifungal effect in vitro and in vivo, respectively. Mechanism studies were investigated by morphogenesis observation, Fluo-3/AM, DCFH-DA and rhodamine6G assay, respectively. Results: CHT exhibited antifungal activity against C. albicans and preformed biofilms with minimum inhibitory concentrations ranged from 2 to 16 μg/ml. Besides, CHT protected G. mellonella larvae infected by C. albicans. Mechanisms studies revealed that CHT inhibited hyphal growth, increased intracellular calcium concentration, induced accumulation of reactive oxygen species and inhibited drug transporter activity. Conclusion: CHT exhibited antifungal activity against C. albicans.

scholarly journals Focal Accumulation of ROS Can Block Pyricularia oryzae Effector BAS4-Expression and Prevent Infection in Rice

2020 ◽  
Vol 21 (17) ◽  
pp. 6196
Author(s):  
Yafei Chen ◽  
Sarmina Dangol ◽  
Juan Wang ◽  
Nam-Soo Jwa
Keyword(s):  
Plant Immunity ◽  
Pathogenic Fungus ◽  
Hyphal Growth ◽  
Infection Process ◽  
Pyricularia Oryzae ◽  
High Concentrations ◽  
Ros Burst ◽  
Immunity Mechanism ◽  
Respiratory Burst Oxidase

The reactive oxygen species (ROS) burst is the most common plant immunity mechanism to prevent pathogen infection, although the exact role of ROS in plant immunity has not been fully elucidated. We investigated the expression and translocation of Oryza sativa respiratory burst oxidase homologue B (OsRBOHB) during compatible and incompatible interactions between rice epidermal cells and the pathogenic fungus Pyricularia oryzae (syn. Magnaporthe oryzae). We characterized the functional role of ROS focal accumulation around invading hyphae during P. oryzae infection process using the OsRBOHB inhibitor diphenyleneiodonium (DPI) and the actin filament polymerization inhibitor cytochalasin (Cyt) A. OsRBOHB was strongly induced during incompatible rice–P. oryzae interactions, and newly synthesized OsRBOHB was focally distributed at infection sites. High concentrations of ROS focally accumulated at the infection sites and suppressed effector biotrophy-associated secreted (BAS) proteins BAS4 expression and invasive hyphal growth. DPI and Cyt A abolished ROS focal accumulation and restored P. oryzae effector BAS4 expression. These results suggest that ROS focal accumulation is able to function as an effective immune mechanism that blocks some effectors including BAS4-expression during P. oryzae infection. Disruption of ROS focal accumulation around invading hyphae enables successful P. oryzae colonization of rice cells and disease development.

scholarly journals Limonin Enhances the Antifungal Activity of Eugenol Nanoemulsion against Penicillium Italicum In Vitro and In Vivo Tests

2021 ◽  
Vol 9 (5) ◽  
pp. 969
Author(s):  
Yi Li ◽  
Runan Zhao ◽  
Yan Li ◽  
Zhiqin Zhou
Keyword(s):  
Antifungal Activity ◽  
Pathogenic Fungus ◽  
Germination Rate ◽  
Challenge Test ◽  
Postharvest Quality ◽  
Citrus Fruits ◽  
Penicillium Italicum ◽  
Blue Mold

Penicillium italicum, the cause of citrus blue mold, is a pathogenic fungus that seriously affects the postharvest quality of citrus fruit and causes serious economic loss. In this study, a eugenol nanoemulsion containing limonin, an antimicrobial component from citrus seeds, was prepared using a high-pressure microfluidizer and the antifungal activity of the nanoemulsions against P. italicum was evaluated based on the conidial germination rate, mycelial growth, and scanning electron microscopy analysis. The results showed that the minimum inhibitory concentration and the inhibition rate of limonin-loaded eugenol nanoemulsion was 160 μg/mL and 59.21%, respectively, which was more potent than that of the limonin-free eugenol emulsion. After treatment with the nanoemulsions, the integrity of the P. italicum cell membrane was disrupted, the cell morphology was abnormal, and the leakage of nucleic acid and protein was observed. In addition, the challenge test on citrus fruits revealed that the limonin-loaded eugenol emulsion inhibited citrus infection for longer periods, with an infection rate of 29.2% after 5 days. The current research shows that nanoemulsions containing limonin and eugenol have effective antifungal activity against P. italicum, and may be used as a substitute for inhibiting blue mold in citrus fruits.

scholarly journals Novel Hit Compounds as Putative Antifungals: The Case of Aspergillus fumigatus

Molecules ◽  
2019 ◽  
Vol 24 (21) ◽  
pp. 3853 ◽  
Author(s):  
Eftichia Kritsi ◽  
Minos-Timotheos Matsoukas ◽  
Constantinos Potamitis ◽  
Anastasia Detsi ◽  
Marija Ivanov ◽  
...  
Keyword(s):  
Antifungal Activity ◽  
Aspergillus Fumigatus ◽  
Fungal Pathogens ◽  
Fungal Infections ◽  
Pharmacophore Model ◽  
Antifungal Drugs ◽  
In Vitro Assays ◽  
Minimum Fungicidal Concentration ◽  
Dynamics Simulations

The prevalence of invasive fungal infections has been dramatically increased as the size of the immunocompromised population worldwide has grown. Aspergillus fumigatus is characterized as one of the most widespread and ubiquitous fungal pathogens. Among antifungal drugs, azoles have been the most widely used category for the treatment of fungal infections. However, increasingly, azole-resistant strains constitute a major problem to be faced. Towards this direction, our study focused on the identification of compounds bearing novel structural motifs which may evolve as a new class of antifungals. To fulfil this scope, a combination of in silico techniques and in vitro assays were implemented. Specifically, a ligand-based pharmacophore model was created and served as a 3D search query to screen the ZINC chemical database. Additionally, molecular docking and molecular dynamics simulations were used to improve the reliability and accuracy of virtual screening results. In total, eight compounds, bearing completely different chemical scaffolds from the commercially available azoles, were proposed and their antifungal activity was evaluated using in vitro assays. Results indicated that all tested compounds exhibit antifungal activity, especially compounds 1, 2, and 4, which presented the most promising minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values and, therefore, could be subjected to further hit to lead optimization.

scholarly journals Single-amino-acid substitutions within the signal sequence of yeast prepro-alpha-factor affect membrane translocation.

1988 ◽  
Vol 8 (5) ◽  
pp. 1915-1922 ◽  
Author(s):  
D S Allison ◽  
E T Young
Keyword(s):  
Amino Acid ◽  
Signal Sequence ◽  
Mitochondrial Protein ◽  
Point Mutations ◽  
Yeast Cells ◽  
Single Amino Acid ◽  
Hydrophobic Core ◽  
Free System ◽  
Alpha Factor

We used a genetic approach to identify point mutations in the signal sequence of a secreted eucaryotic protein, yeast alpha-factor. Signal sequence mutants were obtained by selecting for cells that partially mistargeted into mitochondria a fusion protein consisting of the alpha-factor signal sequence fused to the mature portion of an imported mitochondrial protein (Cox IV). The mutations resulted in replacement of a residue in the hydrophobic core of the signal sequence with either a hydrophilic amino acid or a proline. After reassembly into an intact alpha-factor gene, the substitutions were found to decrease up to 50-fold the rate of translocation of prepro-alpha-factor across microsomal membranes in vitro. Two of three mutants tested produced lower steady-state levels of alpha-factor in intact yeast cells, although the magnitude of the effect was less than that in the cell-free system.

scholarly journals Pseudomonas fluorescens Showing Antifungal Activity against Macrophomina phaseolina, a Severe Pathogenic Fungus of Soybean, Produces Phenazine as the Main Active Metabolite

Biomolecules ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 1728
Author(s):  
Stefany Castaldi ◽  
Marco Masi ◽  
Francisco Sautua ◽  
Alessio Cimmino ◽  
Rachele Isticato ◽  
...  
Keyword(s):  
Antifungal Activity ◽  
Pseudomonas Fluorescens ◽  
Large Scale ◽  
Pathogenic Fungus ◽  
Macrophomina Phaseolina ◽  
Scale Production ◽  
Oh Groups ◽  
Large Scale Production ◽  
Sar Study

Pseudomonas fluorescens 9 and Bacillus subtilis 54, proposed as biofungicides to control Macrophomina phaseolina, a dangerous pathogen of soybean and other crops, were grown in vitro to evaluate their ability to produce metabolites with antifungal activity. The aim of the manuscript was to identify the natural compounds responsible for their antifungal activity. Only the culture filtrates of P. fluorescens 9 showed strong antifungal activity against M. phaseolina. Its organic extract contained phenazine and mesaconic acid (1 and 2), whose antifungal activity was tested against M. phaseolina, as well as Cercospora nicotianae and Colletotrichum truncatum, other pathogens of soybean; however, only compound 1 exhibited activity. The antifungal activity of compound 1 was compared to phenazine-1-carboxylic acid (PCA, 3), 2-hydroxyphenazine (2-OH P, 4), and various semisynthetic phenazine nitro derivatives in order to perform a structure–activity relationship (SAR) study. PCA and phenazine exhibited the same percentage of growth inhibition in M. phaseolina and C. truncatum, whereas PCA (3) showed lower activity against C. nicotianae than phenazine. 2-Hydroxyphenazine (4) showed no antifungal activity against M. phaseolina. The results of the SAR study showed that electron attractor (COOH and NO2) or repulsor (OH) groups significantly affect the antifungal growth, as well as their α- or β-location on the phenazine ring. Both PCA and phenazine could be proposed as biopesticides to control the soybean pathogens M. phaseolina, C. nicotianae, and C. truncatum, and these results should prompt an investigation of their large-scale production and their suitable formulation for greenhouse and field applications.

scholarly journals AKTIVITAS ANTIFUNGI EKSTRAK DAUN PICUNG TERHADAP CENDAWAN Botryodiplodia theobromae PENYEBAB MATI PUCUK BIBIT JABON MERAH Antifungal Activity of Picung Leaf Extracts Against Botryodiplodia theobromae A Dieback Fungus of Jabon Merah Seedling

2017 ◽  
Vol 8 (2) ◽  
pp. 110-115
Author(s):  
L M. Alfin Agushara Bena ◽  
Achmad . ◽  
Syamsul Falah
Keyword(s):  
Antifungal Activity ◽  
Pathogenic Fungus ◽  
Food Poisoning ◽  
Hot Water ◽  
Botryodiplodia Theobromae ◽  
Leaf Extracts ◽  
Vitro Assay ◽  
Methanol Extracts ◽  
Phytochemical Compounds

Invasion of Botryodiplodia theobromae as a dieback fungus on jabon merah (Anthocephalus macrophyllus) seedlings is a crucial issues. This pathogenic fungus needs to be controlled precisely. Phytochemical compounds of plant material is one alternative fungicides. Crude extracts of picung (Pangium edule) leaf in water and methanol solvent known to have antifungal potential. However, its activity against B. theobromae need to be revealed. In vitro assay aimed to measuring antifungal activity of water and methanol extracts of picung leaf on controlling growth of B. theobromae pathogenic fungus. Hot water extraction and maceration in methanol was conducted to obtain crude extract materials. In-vitro antifungal assay of water and methanol extracts of picung leaf was conducted by food poisoning method on Potato Dextrose Agar media. The results showed that water and methanol extracts of pangi leaf be able to suppress the growth of B. theobromae on the first day of observation, with the highest antifungal index reached 100% at concentration of 25mg/mL and 50mg/mL, respectively. This indicates that water and methanol extracts of picung leaf have antifungal activity. Generally, there was a trend of decrease in antifungal index over time.Key words: antifungal activity, Botryodiplodia theobromae, jabon merah, picung

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