Inhibition of Biofilm and Virulence Factors of Candida albicans by Partially Purified Secondary Metabolites of Streptomyces chrestomyceticus Strain ADP4

Background: Despite several advancements in antifungal drug discovery, fungal diseases like Invasive Candidiasis (IC) still remain associated with high rates of morbidity and mortality worldwide. Thus there is an enormous need for anti-Candida drugs. Objective: The main objectives of the work included: 1. To investigate therapeutically significant classes of secondary metabolites produced by S. chrestomyceticus strain ADP4. 2. To investigate and analyze inhibition of significant virulence attributes of C. albicans, such as, biofilm and secretory hydrolytic enzymes by ADP4 secondary metabolites. 3. Mechanistic analysis of probable compounds for their site of action on Secretary Aspartyl Proteinase 3 (Sap3). Methods: Metabolite extract-SDB (MESDB) of S. chrestomyceticus strain ADP4 was fractionated on silica gel column chromatography. Fractions were analyzed for anti-Candida activity by disc diffusion assay. Active fractions were further purified by differential solvent treatment. MIC90 values were determined by broth dilution method. MFC was based on counting viable cells. Inhibition of yeast to hyphae transition and that of production of hydrolytic enzymes were estimated by plate assays. GC-MS of MESDB and Partially Purified Metabolite preparations (PPMs) was done. GRIP docking studies with Sap 3 of C. albicans was done using VLife MDS 4.6 software. Results: Chemical profiling showed that ADP4 secondary metabolites contained alkaloids, flavonoids, polyphenols, terpenoids and triterpenes. The MESDB and the PPMs showed low or no cytotoxicity but were able to effectively contain virulence attributes of Candida pathogen. Docking studies revealed that some of the probable compounds have affinity for aspartic acid residue in Sap3 enzyme of C. albicans. Conclusion: Secondary metabolite of strain ADP4 included important classes of therapeutically important compounds. Their anti-Candida activity was mediated by inhibition of critical virulence factors of the pathogen.

Download Full-text

Hetero-Tricyclic Lead Scaffold as Novel PDE5A Inhibitor for Antihypertensive Activity: In Silico Docking Studies

Current Computer - Aided Drug Design ◽

10.2174/1573409915666190214161221 ◽

2019 ◽

Vol 15 (4) ◽

pp. 318-333

Author(s):

Dipak P. Mali ◽

Neela M. Bhatia

Keyword(s):

In Silico ◽

Docking Study ◽

Docking Studies ◽

Antihypertensive Activity ◽

In Silico Docking ◽

Π Stacking ◽

Π Stacking Interaction ◽

Nitrogen Heteroatom ◽

Inhibitory Potential ◽

Vlife Mds

Objective:To screen the phytochemicals for phosphodiesterase 5A (PDE5A) inhibitory potential and identify lead scaffolds of antihypertensive phytochemicals using in silico docking studies.Methods:In this perspective, reported 269 antihypertensive phytochemicals were selected. Sildenafil, a PDE5A inhibitor was used as the standard. In silico docking study was carried out to screen and identify the inhibiting potential of the selected phytochemicals against PDE5A enzyme using vLife MDS 4.4 software.Results:Based on docking score, π-stacking, H-bond and ionic interactions, 237 out of 269 molecules were selected which have shown one or more interactions. Protein residue Gln817A was involved in H-boding whereas Val782A, Phe820A and Leu804A were involved in π-stacking interaction with ligand. The selected 237 phytochemicals were structurally diverse, therefore 82 out of 237 molecules with one or more tricycles were filtered out for further analysis. Amongst tricyclic molecules, 14 molecules containing nitrogen heteroatom were selected for lead scaffold identification which finally resulted in three different basic chemical backbones like pyridoindole, tetrahydro-pyridonaphthyridine and dihydro-pyridoquinazoline as lead scaffolds.Conclusion:In silico docking studies revealed that nitrogen-containing tetrahydro-pyridonaphthyridine and dihydro-pyridoquinazoline tricyclic lead scaffolds have emerged as novel PDE5A inhibitors for antihypertensive activity. The identified lead scaffolds may provide antihypertensive lead molecules after its optimization.

Download Full-text

Associated microflora of medicinal ferns: biotechnological potentials and possible applications

International Journal of Bioassays ◽

10.21746/ijbio.2016.03.0017 ◽

2016 ◽

Vol 5 (03) ◽

pp. 4927 ◽

Cited By ~ 3

Author(s):

Shubhi Srivastava ◽

Paul A. K.

Keyword(s):

Secondary Metabolites ◽

Growth Promotion ◽

Biological Activities ◽

Hydrolytic Enzymes ◽

In Vitro Production ◽

Wide Range ◽

Negative Effect ◽

Bacteria And Fungi

Plant associated microorganisms that colonize the upper and internal tissues of roots, stems, leaves and flowers of healthy plants without causing any visible harmful or negative effect on their host. Diversity of microbes have been extensively studied in a wide variety of vascular plants and shown to promote plant establishment, growth and development and impart resistance against pathogenic infections. Ferns and their associated microbes have also attracted the attention of the scientific communities as sources of novel bioactive secondary metabolites. The ferns and fern alleles, which are well adapted to diverse environmental conditions, produce various secondary metabolites such as flavonoids, steroids, alkaloids, phenols, triterpenoid compounds, variety of amino acids and fatty acids along with some unique metabolites as adaptive features and are traditionally used for human health and medicine. In this review attention has been focused to prepare a comprehensive account of ethnomedicinal properties of some common ferns and fern alleles. Association of bacteria and fungi in the rhizosphere, phyllosphere and endosphere of these medicinally important ferns and their interaction with the host plant has been emphasized keeping in view their possible biotechnological potentials and applications. The processes of host-microbe interaction leading to establishment and colonization of endophytes are less-well characterized in comparison to rhizospheric and phyllospheric microflora. However, the endophytes are possessing same characteristics as rhizospheric and phyllospheric to stimulate the in vivo synthesis as well as in vitro production of secondary metabolites with a wide range of biological activities such as plant growth promotion by production of phytohormones, siderophores, fixation of nitrogen, and phosphate solubilization. Synthesis of pharmaceutically important products such as anticancer compounds, antioxidants, antimicrobials, antiviral substances and hydrolytic enzymes could be some of the promising areas of research and commercial exploitation.

Download Full-text

Proteomic and Transcriptomic Analyses Provide Novel Insights into the Crucial Roles of Host-Induced Carbohydrate Metabolism Enzymes in Xanthomonas oryzae pv. oryzae Virulence and Rice-Xoo Interaction

Rice ◽

10.1186/s12284-021-00503-x ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

Guichun Wu ◽

Yuqiang Zhang ◽

Bo Wang ◽

Kaihuai Li ◽

Yuanlai Lou ◽

...

Keyword(s):

Gene Expression ◽

Signal Transduction ◽

Amino Acids ◽

Secondary Metabolites ◽

Carbohydrate Metabolism ◽

Virulence Factors ◽

Global Gene Expression ◽

Xanthomonas Oryzae ◽

Oxidation Reduction ◽

Phenylpropanoid Biosynthesis

Abstract Background Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial leaf blight, a devastating rice disease. The Xoo-rice interaction, wherein wide ranging host- and pathogen-derived proteins and genes wage molecular arms race, is a research hotspot. Hence, the identification of novel rice-induced Xoo virulence factors and characterization of their roles affecting rice global gene expression profiles will provide an integrated and better understanding of Xoo-rice interactions from the molecular perspective. Results Using comparative proteomics and an in vitro interaction system, we revealed that 5 protein spots from Xoo exhibited significantly different expression patterns (|fold change| > 1.5) at 3, 6, 12 h after susceptible rice leaf extract (RLX) treatment. MALDI-TOF MS analysis and pathogenicity tests showed that 4 host-induced proteins, including phosphohexose mutase, inositol monophosphatase, arginase and septum site-determining protein, affected Xoo virulence. Among them, mutants of two host-induced carbohydrate metabolism enzyme-encoding genes, ΔxanA and Δimp, elicited enhanced defense responses and nearly abolished Xoo virulence in rice. To decipher rice differentially expressed genes (DEGs) associated with xanA and imp, transcriptomic responses of ΔxanA-treated and Δimp-treated susceptible rice were compared to those in rice treated with PXO99A at 1 and 3 dpi. A total of 1521 and 227 DEGs were identified for PXO99A vs Δimp at 1 and 3 dpi, while for PXO99A vs ΔxanA, there were 131 and 106 DEGs, respectively. GO, KEGG and MapMan analyses revealed that the DEGs for PXO99A vs Δimp were mainly involved in photosynthesis, signal transduction, transcription, oxidation-reduction, hydrogen peroxide catabolism, ion transport, phenylpropanoid biosynthesis and metabolism of carbohydrates, lipids, amino acids, secondary metabolites, hormones, and nucleotides, while the DEGs from PXO99A vs ΔxanA were predominantly associated with photosynthesis, signal transduction, oxidation-reduction, phenylpropanoid biosynthesis, cytochrome P450 and metabolism of carbohydrates, lipids, amino acids, secondary metabolites and hormones. Although most pathways were associated with both the Δimp and ΔxanA treatments, the underlying genes were not the same. Conclusion Our study identified two novel host-induced virulence factors XanA and Imp in Xoo, and revealed their roles in global gene expression in susceptible rice. These results provide valuable insights into the molecular mechanisms of pathogen infection strategies and plant immunity.

Download Full-text

Antibacterial properties in-vitro of Mexican serviceberry extracts against dental biofilm species

Journal of Berry Research ◽

10.3233/jbr-210718 ◽

2021 ◽

pp. 1-16

Author(s):

Erika-Alejandra Salinas-Peña ◽

Martha Mendoza-Rodríguez ◽

Claudia Velázquez-González ◽

Carlo Eduardo Medina-Solis ◽

América Patricia Pontigo-Loyola ◽

...

Keyword(s):

Secondary Metabolites ◽

Antibacterial Properties ◽

Fusobacterium Nucleatum ◽

Positive Control ◽

Dilution Method ◽

Periodontal Pathogens ◽

Leaf Extracts ◽

Dental Biofilm ◽

Biofilm Bacteria

BACKGROUND: The Mexican serviceberry, Malacomeles denticulata, have been used as a successful oral therapy by Mexican communities without enough scientific support. OBJECTIVE: To evaluate the M. denticulata extracts with selective antibacterial properties over dental biofilm bacteria. METHODS: Fruit, Leaf, and Stem of M. denticulata extracts were evaluated with micro-broth dilution method using ATCC bacteria. OD600 values had compared against each positive control (T-student-test). Anaerobically viability had confirmed by Colony-Forming-Units. Thin-Layer-Chromatography was used to identify the number of compounds and phytochemicals to identify secondary metabolites of the selected extracts. RESULTS: Streptococcus mutans showed Minimum-Bactericidal-Concentrations_(MBC) at 30 mg/mL to Fruit, Leaf, and Stem extracts. Periodontal-pathogens Aggregatibacter actinomycetemcomitans serotype b_(MBC = 30 mg/mL_p < 0.01); Fusobacterium nucleatum subsp. nucleatum_(MBC = 30 mg/mL_p<0.001); Parvimonas micra_(MBC = 15 mg/mL_NS); Porphyromonas gingivalis_(MBC = 30 mg/mL_NS); and Prevotella intermedia_(MBC = 3.75 mg/mL_NS) presented higher sensitivity to Leaf-Methanol, than the primary colonizers. Phytochemicals showed positive results to anthraquinones, coumarins, flavonoids, saponins, saponins steroids/triterpenoids, steroids/triterpenes, and tannins/phenols. CONCLUSION: We suggest the natural extracts of fruit and leaf of the Mexican serviceberry for the preventive use over the oral cariogenic or periodontal biofilm species, by their selective antibacterial properties against pathogenic species evaluated in-vitro, and due to the presence of antibacterial secondary metabolites identified as flavonoids and saponins of M. denticulata leaf extracts.

Download Full-text

Antimicrobial Activities in Pistacia atlantica—Aphids Make a Difference!

Proceedings ◽

10.3390/proceedings2020066009 ◽

2020 ◽

Vol 66 (1) ◽

pp. 9

Author(s):

Yoram Gerchman

Keyword(s):

Secondary Metabolites ◽

Essential Oils ◽

Organic Solvents ◽

Antimicrobial Properties ◽

Antimicrobial Activities ◽

Disk Diffusion ◽

Pistacia Atlantica ◽

Disk Diffusion Assay ◽

Bacteria And Fungi ◽

Diffusion Assay

Plants have been explored and used as sources for antimicrobial extract and compounds for many years, but galls—specialized structures forms on such by diversity of organisms—have been explored much less. Aphid galls host many insects in closed, humid and sugar rich environments for long periods. We have tested the antimicrobial properties of Slavum wertheimae aphid galls on Pistacia atlantica. Secondary metabolites were extracted from leaves and galls with organic solvents, and essential oils with Clevenger, and tested by disk diffusion assay and volatile effect on bacteria and fungi, respectively. The results demonstrated that gall extracts/essential oils had much stronger activity against the diversity of bacteria and fungi. The large diversity of galls suggest they could be explored as source for novel compounds.

Download Full-text

Virulence of Clinical Candida Isolates

Pathogens ◽

10.3390/pathogens10040466 ◽

2021 ◽

Vol 10 (4) ◽

pp. 466

Author(s):

Martyna Mroczyńska ◽

Anna Brillowska-Dąbrowska

Keyword(s):

Virulence Factors ◽

Biofilm Formation ◽

Galleria Mellonella ◽

Hydrolytic Enzymes ◽

Morphological Transition ◽

Candida Spp

The factors enabling Candida spp. infections are secretion of hydrolytic enzymes, adherence to surfaces, biofilm formation or morphological transition, and fitness attributes. The aim of this study was to investigate the correlation between known extracellular virulence factors and survival of Galleria mellonella larvae infected with clinical Candida. The 25 isolates were tested and the activity of proteinases among 24/24, phospholipases among 7/22, esterases among 14/23, hemolysins among 18/24, and biofilm formation ability among 18/25 isolates was confirmed. Pathogenicity investigation using G. mellonella larvae as host model demonstrated that C. albicans isolates and C. glabrata isolate were the most virulent and C. krusei isolates were avirulent. C. parapsilosis virulence was identified as varied, C. inconspicua were moderately virulent, and one C. palmioleophila isolate was of low virulence and the remaining isolates of this species were moderately virulent. According to our study, virulence of Candida isolates is related to the expression of proteases, hemolysins, and esterases.

Download Full-text

IN SILICO STUDIES OF THE SECONDARY METABOLITES OF SOLANUM TORVUM SW. FOR THEIR ANTIASTHMATIC ACTIVITY

International Journal of Current Pharmaceutical Research ◽

10.22159/ijcpr.2017v9i4.20759 ◽

2017 ◽

Vol 9 (4) ◽

pp. 38

Author(s):

Soorya R. ◽

Dhamodaran P. ◽

Rajesh Kumar R. ◽

Duraisamy B.

Keyword(s):

Methyl Ester ◽

Secondary Metabolites ◽

Ethyl Ester ◽

In Silico ◽

Methanolic Extract ◽

Acid Ethyl Ester ◽

Docking Studies ◽

Receptor Protein ◽

Solanum Torvum ◽

In Silico Studies

Objective: Solanum torvum Sw., Family: Solanaceae, commonly known as Turkey Berry is used by the traditional tribes for the treatment of cold, cough, tuberculosis, hepatotoxicity, cancer, etc. The action of the plant towards the treatment of these diseases has been proven except for asthma. The present study is to prove the antiasthmatic activity of methanolic extract and the secondary metabolites of Solanum torvum Sw using in silico docking studies in compare to reference standard Dexamethasone, a synthetic cortisone derivative.Methods: The GC-MS analysis of the dried methanolic extract of the dried fruits of Solanum torvum Sw. and the total saponin fraction has been carried out to know the important moieties that are responsible for the antiasthmatic activity.Results: The results from the docking studies showed that the compounds Cholesta-5,7,9-(11)-trien-3-ol,4,4-dimethyl, (3á); Lanosta-7,9-(11),20-triene-3α, 18-diol, diacetate and Cholestan-26-oic acid,3,7,12,24-tetrakis (acetyloxy), methyl ester, (3à,5á,7à,12à) were found to have significant scores of-6.8,-6.9 and-6.9 respectively towards Glucocorticoid receptor protein (Gr), (PDB id: 4UDC) which is very similar to the affinity of the standard (-7.1). These compounds passed the drug-likeness test. A modification in the structure can be brought, which makes the compounds more potent. The compounds 9, 12-Octadecadienoic acid, ethyl ester; Hexadecanoic acid, ethyl ester; 9-Octadecenoic acid (Z), methyl ester; Oxacycloheptadec-8-en-2-one, (8Z) have passed the Blood Brain Barrier (BBB) filter of the drug-likeness test.Conclusion: The antiasthmatic activity of the drug may be due to the similarity with the structure of Dexamethasone. Further research can be carried out in order to improve the clinical significance of these extracts and its metabolites.

Download Full-text

Qualitatively and quantitatively comparing secondary metabolites in three medicinal parts derived from Poria cocos (Schw.) Wolf using UHPLC-QTOF-MS/MS-based chemical profiling

Journal of Pharmaceutical and Biomedical Analysis ◽

10.1016/j.jpba.2017.11.066 ◽

2018 ◽

Vol 150 ◽

pp. 278-286 ◽

Cited By ~ 18

Author(s):

Lixia Zhu ◽

Jun Xu ◽

Shijin Zhang ◽

Rujing Wang ◽

Qing Huang ◽

...

Keyword(s):

Secondary Metabolites ◽

Poria Cocos ◽

Chemical Profiling

Download Full-text

Ginseng aqueous extract attenuates the production of virulence factors, stimulates twitching and adhesion, and eradicates biofilms of Pseudomonas aeruginosa

Canadian Journal of Physiology and Pharmacology ◽

10.1139/y11-057 ◽

2011 ◽

Vol 89 (6) ◽

pp. 419-427 ◽

Cited By ~ 12

Author(s):

Misagh Alipour ◽

Abdelwahab Omri ◽

Zacharias E. Suntres

Keyword(s):

Pseudomonas Aeruginosa ◽

Virulence Factors ◽

Aqueous Extract ◽

North American ◽

American Ginseng ◽

Antimicrobial Activities ◽

Live Cells ◽

Dilution Method ◽

Agar Dilution Method

This study was carried out to examine the antimicrobial activity of the aqueous extract of Panax quinquefolius from North American ginseng (NAGE) root against Pseudomonas aeruginosa . The minimum inhibitory concentrations of reference and clinical isolates of Pseudomonas aeruginosa were measured by a standard agar-dilution method. At subinhibitory NAGE concentrations, the secretion of virulence factors, motility on agar, and adhesion to 96-well microplates were studied on the nonmucoid Pseudomonas aeruginosa O1 strain. At suprainhibitory concentrations, the activity of NAGE against mature biofilm complexes formed in the Calgary Biofilm Device and the Stovall flow cell were assessed. NAGE possessed an antibacterial activity against all the Pseudomonas aeruginosa strains at 1.25%–2.5% w/v. NAGE also significantly attenuated pyocyanin, pyoverdine, and lipase concentrations, stimulated twitching, and attenuated swarming and swimming motility. At 1.25% w/v, NAGE augmented adhesion, and at 5% w/v detached 1-day-old biofilms in microplates. The extract also eradicated 6-day-old mature biofilms (5% w/v), and fluorescence microscopy displayed a reduction of live cells and biofilm complexes compared with nontreated biofilms. These data suggest that the aqueous extract from North American ginseng possesses antimicrobial activities in vitro.

Download Full-text