Microbial Extracts Research Articles

Pathological Characterization and Management of Lasiodiplodia theobromae, a Hemibiotroph with an Interkingdom Host Range.

Heart rot disease, caused by Lasiodiplodia theobromae, is destructive for date palms and other woody plants. The disease was reported in several oases in Egypt, and the pathogen was found in association with infected trees suffering dieback and rachis blight. Seven phylogenetically distinct fungal isolates were selected, and their pathogenicity was confirmed on date palms. The isolates exhibited variable degrees of virulence on inoculated leaves, which confirms the variation. We examined the antifungal effect of microbial bioagents and plant extracts on heart rot disease. The isolates of Trichoderma spp. gave moderate reduction of the pathogen's linear growth (40 to 60%), whereas their exudates were ultimately ineffective. Bacillus spp. isolates, except for B. megaterium, were more effective against spore germination, giving 80 to 90% reduction on average. Among the examined plant extracts, garlic sap gave 98.67% reduction of linear growth followed by artemisia (15.5%) and camphor (24.8%). The extraction methods greatly influenced the antifungal efficiency of each extract because exposure to organic solvents significantly decreased the efficiency of all extracts, whereas hot water extraction negatively affected garlic sap only. Successful bioagents and plant extracts were further assayed for the suppression of heart rot disease on date palms. Both T. album and T. harzianum gave comparable degrees of suppression as by commercial fungicides. In addition, treatment before or during pathogen inoculation was the most effective because it significantly enhanced the expression of defense-related enzymes. Our findings suggest biopesticides possess a dual role in disease suppression and defense boosters for date palms suffering heart rot disease.

Antibacterial activity and antioxidant capacity of dairy kefir beverages

Kefir traditionally is based on pasteurised milk and the use of other substrates is unusual. Kefir beverages made with raw milk (RMK), pasteurised milk (PMK) or whey (WK), were obtained and fractionated in microbial cells and supernatants. Antioxidant and antibacterial activity of fractions against Cronobacter sakazakii and Escherichia coli were analysed. The highest antioxidant activity was obtained for PMK with values between 8500 and 10 000 μm TE/mg. Microbial extracts from RMK and PMK showed a reduction of C. sakazakii of 7 log units; while microbial extracts and supernatants from RMK decreased E. coli in 3 and 5 log units, respectively. These results indicate that dairy kefir beverages can be an excellent source of defensive agents to fight against bacteria that cause gastrointestinal disorders.

Influence of Heat Treatment and Lactic Acid Fermentation on the Physical and Chemical Properties of Pumpkin Juice.

Pumpkin is a highly nutritious plant, rich in valuable nutrients that benefit human health. Due to the high perishability of this fruit, the production of pumpkin juice is a practical way to use it effectively. Recently, fermented vegetable juices have been used as a dairy alternative due to their nutritional and potential probiotic properties. This study investigated the fermentation of pumpkin juice using different strains of lactic acid bacteria (LAB), with and without heat treatment. The effects of fermentation on microbial growth, pH, acidity, extract, sugars, carotenoids, polyphenols, and antioxidant properties were analyzed. The heat-treatment process did not greatly impact the dry matter content, pH, acidity, extract, or sugar content. However, it led to a reduction in carotenoid and polyphenol levels. During fermentation, there was a consistent decrease in pH and an increase in total acidity, with no noticeable differences between bacterial strains regarding their influence on these parameters. The study revealed that there were no distinctions between LAB strains in their effects on pH, acidity, and carotenoid content in fermented pumpkin juice. Nonetheless, both L. sakei and L. plantarum proved to be effective in the fermentation process, with L. sakei demonstrating greater adaptability. The expected pH, acidity, and sugar content changes were consistently observed throughout the fermentation process. Overall, results confirm the efficacy of the used Lactobacillus strains in fermenting pumpkin juice and highlight the potential impact of heat treatment on the nutritional composition of the juice. The purpose of thermal processing of pumpkin juice, which is conducted with lactic acid fermentation, is crucial for the food industry. It extends the product's shelf life, improves its nutritional and taste profiles, and guarantees its microbiological safety.

Biosynthesis and Characterization of Zinc Nanoparticles Using Strains of Pseudomonas and Actinobacteria

Research was carried out at the Green Nanotechnology Laboratory, University of Agricultural Sciences, Dharwad, Karnataka, with a specific emphasis on the biosynthesis of zinc nanoparticles using Pseudomonas and Actinobacteria. Collection and screening of Pseudomonas and Actinobacterial isolates were done at Microbial Genetics Laboratory, Department of Agricultural Microbiology, UAS, Dharwad. The ZnNPswere biosynthesized and characterized through UV-Visible spectroscopy, Particle Size Analyzer (PSA), Scanning Electron Microscope (SEM), Energy-dispersive X-ray spectroscopy (EDS), X-Ray Diffraction (XRD) and Fourier Transform Infrared Spectroscopy (FTIR). The ZnNPs synthesized through Pseudomonas(AUDP209) and Actinobacteria(AUDT636)isolates exhibited peak values at 365 and 374 nm, with average sizes of 48.7 nm and 54.9 nm, respectively.SEM images revealed the spherical shape of the biosynthesized ZnNPs, while EDX confirmed the zinc ions presence in the biosynthesized samples and the crystalline structure of the ZnNPswas confirmed by XRD.FTIR spectroscopy confirmed the organic compounds present in the microbial extracts responsible for the capping and stabilizing agents for biosynthesis of ZnNPs.

Force-enhanced sensitive and specific detection of DNA-intercalative agents directly from microorganisms at single-molecule level.

Microorganisms can produce a vast array of bioactive secondary metabolites, including DNA-intercalating agents like actinomycin D, doxorubicin, which hold great potential for cancer chemotherapy. However, discovering novel DNA-intercalating compounds remains challenging due to the limited sensitivity and specificity of conventional activity assays, which require large-scale fermentation and purification. Here, we introduced the single-molecule stretching assay (SMSA) directly to microbial cultures or extracts for discovering DNA-intercalating agents, even in trace amounts of microbial cultures (5 μl). We showed that the unique changes of dsDNA incontour length and overstretching transition enable thespecific detection of intercalators from complex samples without the need forextensive purification. Applying force to dsDNA also enhanced the sensitivity by increasing both the binding affinity Ka and the quantity of ligands intercalation, thus allowing thedetection of weak intercalators, which are often overlooked using traditional methods. We demonstrated the effectiveness of SMSA,identified two DNA intercalator-producing strains: Streptomyces tanashiensis and Talaromyces funiculosus, and isolated three DNA intercalators: medermycin, kalafungin and ligustrone B. Interestingly, both medermycin and kalafungin, classified as weak DNA intercalators (Ka ∼103 M-1), exhibited potent anti-cancer activity against HCT-116 cancer cells, with IC50 values of 52±6and 70±7 nM, respectively.

Intracellular pyruvate as one of the major bioactive substances of lactic acid bacteria isolated from kimchi.

The present study aimed to identify the metabolites associated with the physiological activity of kimchi-derived lactic acid bacteria (LAB). A clear difference was observed between the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical scavenging rates when the pyruvate content was high (273.5 ng/µL; radical removal speed 6.50% per min) and the rates when the pyruvate content had decreased (131.9 ng/µL; radical removal speed 3.63% per min). Additionally, the characteristics of LAB antioxidant activity (increase in ABTS radical scavenging activity with reaction time, low level of 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity) were similar to those of pyruvate-derived activity. Hydrogen peroxide content (WiKim0124, 2.08 → 0.26; WiKim0121, 0.99 → 0.47; WiKim39, 1.93 → 0.24) and lactate dehydrogenase activity (WiKim0124, 1.53 → 0.00; WiKim0121, 0.73 → 0.01; WiKim39, 1.72 → 0.02) decreased more in heat-killed LAB than in non-heat-killed LAB. Accordingly, this resulted in increased pyruvate content and the inhibitory activity of lipid peroxide production increased by 2-3 times. Our findings indicate that pyruvate is one of the major metabolites regulating LAB physiological activity. PRACTICAL APPLICATION: The safety of utilizing live probiotics remains a topic of debate. To mitigate associated risks, there is a growing interest in non-viable microorganisms or microbial cell extracts for use as probiotics. Various methods can be employed for probiotic inactivation. Heat treatment typically emerges as the preferred choice for inactivating probiotic strains in many instances. The present study shows the distinctions between inactivating lactic acid bacteria (LAB) through heat treatment and non-heat treatment. It may serve as a valuable reference for selecting an appropriate inactivation method for LAB in industrial processes.

Identification of novel small molecule-based strategies of COL7A1 upregulation and readthrough activity for the treatment of recessive dystrophic epidermolysis bullosa

Recessive dystrophic epidermolysis bullosa (RDEB) is a rare genetic disease caused by loss of function mutations in the gene coding for collagen VII (C7) due to deficient or absent C7 expression. This disrupts structural and functional skin architecture, leading to blistering, chronic wounds, inflammation, important systemic symptoms affecting the mouth, gastrointestinal tract, cornea, and kidney function, and an increased skin cancer risk. RDEB patients have an extremely poor quality of life and often die at an early age. A frequent class of mutations in RDEB is premature termination codons (PTC), which appear in homozygosity or compound heterozygosity with other mutations. RDEB has no cure and current therapies are mostly palliative. Using patient-derived keratinocytes and a library of 8273 small molecules and 20,160 microbial extracts evaluated in a phenotypic screening interrogating C7 levels, we identified three active chemical series. Two of these series had PTC readthrough activity, and one upregulated C7 mRNA, showing synergistic activity when combined with the reference readthrough molecule gentamicin. These compounds represent novel potential small molecule-based systemic strategies that could complement topical-based treatments for RDEB.

Development of innovative dye sensitized solar cells (DSSCs) based on co-sensitization of natural microbial pigments

In the last years, the growing attention towards environmental sustainability and circular economy has led to a renewed interest in the use of eco-friendly and recyclable materials in various sectors.Developing innovative dye sensitized solar cells (DSSCs) based on microbial pigments, is very important to meet the demands of sustainable devices. Microbial extracts obtained from Talaromyces atroroseus GH2, Arthrobacter bussei CP30 and Paracoccus bogoriensis BOG6 cultivations, and characterized by HPLC-DAD-ESI-MS analyses, have been used in this work for this purpose. The extracted pigments were tested to evaluate their suitability as photosensitizers through co-sensitization method. UV–vis measurements were carried out to determine the absorbance intensity, while Photoelectrochemical and Electrochemical Impedance Spectroscopy (EIS) analyses were applied to evaluate the devices' photovoltaic parameters and impedance characteristics. The best device, obtained by the co-sensitization of the dyes produced by Talaromyces atroroseus GH2/Paracoccus bogoriensis BOG6, exhibited a Jsc of 1.59 mA/cm2, Voc of 0.35 V, FF of 0.62, and a PCE of 0.34 %. This study highlights the potential of microbial-derived pigments in the development of DSSCs.

Diverse bioactive secondary metabolites from Aspergillus terreus: antimicrobial, anticancer, and anti-SARS-CoV-2 activity studies.

Owing to its high interest as prolific source of diverse bioactive compounds referred in our previous research work, we have scaled-up the fermentation of themarine Aspergillus terreus LGO13 on a liquid culture medium to isolate and identify the very minor/further promising bioactive secondary metabolites and to study their antibacterial, cytotoxic, and antiviral properties. Twenty-three known bioactive metabolites, including the recently discovered microbial natural product N-benzoyl-tryptophan (1), were obtained herein. Their structures were determined using HR-ESI-MS 1D/2D NMR spectroscopy and data from the literature. The biological properties of the microbial extract and the resulting compounds were examined using a set of microorganisms, cervix carcinoma KB-3-1, nonsmall cell lung cancer (NSCLC) A549, and coronavirus (SARS-CoV-2), respectively. Molecular docking (MD) simulations were used to investigate the potential targets of the separated metabolites as anti-SARS-CoV-2 drugs. According to the current study, a viral protein that may be the target of anticovid drugs is a papain-like protease (PLpro), and chaetominine (2) appears to be a viable choice against this protein. We evaluated the antiviral efficacy of chaetominine (2), fumitremorgin C (6), and azaspirofuran A (9) against SARS-CoV-2 based on MD data. Chaetominine (2) and azaspirofuran A (9) displayed intermediate selectivity indices (SI=6.6 and 3.2, respectively), while fumitremorgin C (6) displayed a high selectivity index (SI=19.77). These findings show that fumitremorgin C has promising antiviral action against SARS-CoV-2.

Onychocolone A produced by the fungus Onychocola sp. targets cancer stem cells and stops pancreatic cancer progression by inhibiting MEK2-dependent cell signaling

Pancreatic cancer (PC) shows a high fatality rate that can only be faced with a combination of surgery and chemotherapy or palliative treatment in the case of advanced patients. Besides, PC tumors are enriched with subpopulations of cancer stem cells (CSCs) that are resistant to the existing chemotherapeutic agents, which raises an important need for the identification of new drugs. To fill this gap, we have tested the anti-tumoral activity of microbial extracts, which chemical diversity offers a broad spectrum of potential new bioactive compounds. Extracts derived from the fungus Onychocola sp. CF-107644 were assayed via high throughput screening followed by bioassay-guided fractionation and resulted in the identification and isolation of six benzophenone derivatives with antitumoral activity: onychocolones A-F (#1-6). The structures of the compounds were established by spectroscopic methods, including ESI-TOF MS, 1D and 2D NMR analyses and X-ray diffraction. Compounds #1-4 significantly inhibited the growth of the pancreas tumoral cell lines, with low-micromolar Median Effective Doses (ED50s). Compound #1 (onychocolone A) was prioritized for further profiling due to its pro-apoptotic effect, which was further validated on 3D spheroids and pancreatic CSCs. Protein expression assays showed that the effect was mechanistically linked to the inhibition of MEK onco-signaling pathway. The efficacy of onychocolone A was also demonstrated in vivo by the reduction of tumor growth in a pancreatic xenograft mouse model generated by CSCs. Altogether, the data support that onychocolone A is a promising new small molecule for hit-to-lead development of a new treatment for PC.

Synergistic Effects of Chemical Fungicides with Crude Extracts from Bacillus amyloliquefaciens to Control Northern Corn Leaf Blight

In this research, our objective was to investigate the combined impact of microbial extracts and chemical fungicides on Northern corn leaf blight (NCLB), which is induced by Exserohilum turcicum, and the growth-promoting effect of the crude extracts was also determined. NCLB poses a serious threat to global maize production, necessitating sustainable and environmentally friendly solutions. Mycelial growth rate assays were used to assess the single or synergistic effects of microbial crude extracts and chemical fungicides, and the seed-soaking and root irrigation method was used to detect the growth-promoting effect of the crude extracts on maize seedlings. The results revealed an 84.60% inhibition rate of B. amyloliquefaciens gfj-4 against E. turcicum, and with an EC50 of 49.01 mg·L−1 for the crude extracts. Chemical fungicides demonstrated varying toxicity levels, with fludioxonil exhibiting the highest potency. The mixture of the crude extracts and pyraclostrobin at an 8:2 volume ratio displayed the highest toxicity ratio of 1.24, indicating a synergistic effect. The selected combinations exhibited strong synergistic effects. Soaking maize seeds with 80 mg·L−1 of the crude extracts followed by root irrigation with 40 mg·L−1 produced the most significant growth-promoting effect on maize seedlings. This study highlights the potential of microbial crude extracts to enhance the control of NCLB when combined with pyraclostrobin, along with its growth-promoting effects on maize seedlings.

Antarctic marine sediment as a source of filamentous fungi-derived antimicrobial and antitumor compounds of pharmaceutical interest.

Antarctica harbors a microbial diversity still poorly explored and of inestimable biotechnological value. Cold-adapted microorganisms can produce a diverse range of metabolites stable at low temperatures, making these compounds industrially interesting for biotechnological use. The present work investigated the biotechnological potential for antimicrobial and antitumor activity of filamentous fungi and bacteria isolated from marine sediment samples collected at Deception Island, Antarctica. A total of 89 microbial isolates were recovered from marine sediments and submitted to an initial screening for L-glutaminase with antitumoral activity and for antimicrobial metabolites. The isolates Pseudogymnoascus sp. FDG01, Pseudogymnoascus sp. FDG02, and Penicillium sp. FAD33 showed potential antiproliferative action against human pancreatic carcinoma cells while showing no toxic effect on non-tumor cells. The microbial extracts from unidentified three bacteria and four filamentous fungi showed antibacterial activity against at least one tested pathogenic bacterial strain. The isolate FDG01 inhibited four bacterial species, while the isolate FDG01 was active against Micrococcus luteus in the minimal inhibitory concentration of 0.015625μgmL-1. The results pave the way for further optimization of enzyme production and characterization of enzymes and metabolites found and reaffirm Antarctic marine environments as a wealthy source of compounds potentially applicable in the healthcare and pharmaceutical industry.

The stress history of soil bacteria under organic farming enhances the growth of wheat seedlings.

The effects of stress factors associated with climate change and agricultural management practices on microorganisms are often studied separately, and it remains to be determined how these factors impact the soil microbiome and, subsequently, plant growth characteristics. The aim of this study was to understand how the historical climate and agriculture to which soil microbes have been exposed can influence the growth characteristics of wheat seedlings and their associated bacterial communities. We collected soil from organic and conventional fields with different histories of climate conditions to extract microbes to inoculate wheat seeds under agar-based cultivation conditions. Within a growth period of 8 days, we monitored germination rates and time as well as seedling above-ground biomass and their associated bacterial communities. The results showed a positive interaction between conventional farming practices and an ambient climate for faster and higher germination rates. We demonstrate that soil microbial extracts from organic farming with experience of the future climate significantly enhanced above-ground biomass along with the diversity of bacterial communities associated with seedlings than other treatments. Such findings support the idea that organic agricultural practices not only mitigate the adverse effects of climate change but also promote the diversity of seedling-associated bacteria.

Discovery of bioactive natural products of microbial origin as inhibitors of the PD-1/PD-L1 protein-protein interaction

The PD-1/PD-L1 protein-protein interaction (PPI) controls an adaptive immune resistance mechanism exerted by tumor cells to evade immune responses. The large-molecule nature of current commercial monoclonal antibodies against this PPI hampers their effectiveness by limiting tumor penetration and inducing severe immune-related side effects. Synthetic small-molecule inhibitors may overcome such limitations and have demonstrated promising clinical translation, but their design is challenging. Microbial natural products (NPs) are a source of small molecules with vast chemical diversity that have proved anti-tumoral activities, but which immunotherapeutic properties as PD-1/PD-L1 inhibitors had remained uncharacterized so far. Here, we have developed the first cell-based PD-1/PD-L1 blockade reporter assay to screen NPs libraries. In this study, 6000 microbial extracts of maximum biosynthetic diversity were screened. A secondary metabolite called alpha-cyclopiazonic acid (α-CPA) of a bioactive fungal extract was confirmed as a new PD-1/PD-L1 inhibitor with low micromolar range in the cellular assay and in an additional cell-free competitive assay. Thermal denaturation experiments with PD-1 confirmed that the mechanism of inhibition is based on its stabilization upon binding to α-CPA. The identification of α-CPA as a novel PD-1 stabilizer proves the unprecedented resolution of this methodology at capturing specific PD-1/PD-L1 PPI inhibitors from chemically diverse NP libraries.

Antimicrobial activity of spiculisporic acid isolated from endophytic fungus Aspergillus cejpii of Hedera helix against MRSA

The surge in multidrug-resistant pathogens worldwide has jeopardized the clinical efficiency of many current antibiotics. This problem steered many researchers in their quest to discover new effective antimicrobial agents from natural origins including plants or their residing endophytes. In this work, we aimed to identify the endophytic fungi derived from Hedera helix L. and investigate their potential antimicrobial activity. Bioguided fractionation approach was conducted to isolate the pure compounds from the most active fungal fraction. Out of a total of six different isolated endophytic fungal strains, only Aspergillus cejpii showed the highest activity against all tested microbial strains. The most active fraction was the dichloromethane/methanol fraction (DCM:MeOH), where it showed significant activity against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Serratia marcescens, Acinetobacter baumannii, Salmonella typhi, and three drug-resistant clinical isolate strains including Methicillin-resistant Staphylococcus aureus (MRSA, H1), Pseudomonas aeruginosa (PS 16), and Acinetobacter baumannii (ACT 322) using tetracyline and kanamycin as the control antibiotics. Bioguided fractionation of the active fraction led to the isolation of the γ-butenolide, spiculisporic acid. Structure elucidation was carried out using 1H and 13C-NMR spectroscopic analysis. The compound showed good antimicrobial activities with minimum inhibitory concentration (MIC) values ranging from 3.9 to 31.25 μg/mL against all tested strains. Gas chromatography coupled to mass spectrometry (GC-MS) profiling was also carried out to identify the metabolites in the microbial crude extract. In conclusion, endophytic fungi, Aspergillus cejpii, isolated from Hedera helix L. roots showed promising antimicrobial activity which merits further in-depth investigations for potential utilization as a source of new antibiotics in the future. It can also be considered as a novel source for spiculisporic acid.

Concentration and pH Influence on the Reducing Property of Silk Fibroin from Silk Fibrous Waste in the Green Synthesis of Silver Nanoparticles

The development of methods of green synthesis of metal nanoparticles from the use of biological agents, and microbial or plant extracts, allows the obtainment of biocompatible nanostructures by reducing or eliminating the use of toxic substances. On that basis, this research focused on evaluating the influence of silk fibroin from silk fibrous waste (SFw) on the synthesis of silver nanoparticles (AgNPs), by varying the concentrations of the precursor and reducing agent, as well as the change in the pH value of the reaction. The nanostructures were characterized by UV‐visible spectrophotometry, scanning transmission electron microscopy (STEM) and transmission electron microscopy (TEM), zeta potential, and Fourier transform infrared spectroscopy (FTIR) with an ATR module. The results showed that SFw behaves as a reducing matrix and micellar structure, which favored the formation and obtainment of silver nanoparticles with colloidal stability, particle diameters, and defined morphologies. These aspects were evidenced through the identification of the characteristic surface plasmon resonance of the silver ion, the determination of the particle diameter, the electrokinetic potential, and the identification of the functional groups because of the oxidation of the reducing agent.

Biosynthesis of selenium nanoparticles using cell-free extract of Xenorhabdus cabanillasii GU480990 and their potential mosquito larvicidal properties against yellow fever mosquito Aedes aegypti

Nanomaterials are successful due to their numerous applications in various domains such as cancer treatment, environmental applications, drug and gene delivery. Selenium is a metalloid element with broad biological activities and low toxicity especially at the nanoscale. Several studies have shown that nanoparticles synthesized from microbial and plant extracts are effective against important pests and pathogens. This study describes the bio fabrication of selenium nanoparticles using cell free extract of Xenorhabdus cabanillasii (XC-SeNPs) and assessed their mosquito larvicidal properties. Crystallographic structure and size of XC-SeNPs were determined with UV-a spectrophotometer, Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction analysis (XRD), Energy-dispersive X-ray spectroscopy (EDAX), Zeta potential and Transmission electron microscopy (TEM). The significant surface plasmon resonance at 275 nm indicated the synthesis of XC-SeNPs from the pure cell-free extract of X. cabanillasii. The XRD result exhibits the crystalline nature of XC-SeNPs. The Zeta potential analysis confirmed that the surface charge of XC-SeNPs was −24.17 mV. TEM analysis revealed that synthesized XC-SeNPs were monodispersed, spherically shaped, and sized about 80–200 nm range. In addition, the larvicidal potentials of the bio-fabricated XC-SeNPs were assessed against the 4th-instar Ae. aegypti. XC-SeNPs displayed a dose-dependent larvicidal effect; the larval mortality was 13.3 % at the minimum evaluated concentration and increased to 72 % at higher dose treatments. The LC50 and LC90 concentration of XC-SeNPs against mosquito larvae were 79.4 and 722.4 ppm, respectively.

Pigmented Microbial Extract (PMB) from Exiguobacterium Species MB2 Strain (PMB1) and Bacillus subtilis Strain MB1 (PMB2) Inhibited Breast Cancer Cells Growth In Vivo and In Vitro.

Breast cancer (BC) continues to be one of the major causes of cancer deaths in women. Progress has been made in targeting hormone and growth factor receptor-positive BCs with clinical efficacy and success. However, little progress has been made to develop a clinically viable treatment for the triple-negative BC cases (TNBCs). The current study aims to identify potent agents that can target TNBCs. Extracts from microbial sources have been reported to contain pharmacological agents that can selectively inhibit cancer cell growth. We have screened and identified pigmented microbial extracts (PMBs) that can inhibit BC cell proliferation by targeting legumain (LGMN). LGMN is an oncogenic protein expressed not only in malignant cells but also in tumor microenvironment cells, including tumor-associated macrophages. An LGMN inhibition assay was performed, and microbial extracts were evaluated for in vitro anticancer activity in BC cell lines, angiogenesis assay with chick chorioallantoic membrane (CAM), and tumor xenograft models in Swiss albino mice. We have identified that PMB from the Exiguobacterium (PMB1), inhibits BC growth more potently than PMB2, from the Bacillus subtilis strain. The analysis of PMB1 by GC-MS showed the presence of a variety of fatty acids and fatty-acid derivatives, small molecule phenolics, and aldehydes. PMB1 inhibited the activity of oncogenic legumain in BC cells and induced cell cycle arrest and apoptosis. PMB1 reduced the angiogenesis and inhibited BC cell migration. In mice, intraperitoneal administration of PMB1 retarded the growth of xenografted Ehrlich ascites mammary tumors and mitigated the proliferation of tumor cells in the peritoneal cavity in vivo. In summary, our findings demonstrate the high antitumor potential of PMB1.

Epiphytic and endophytic microbiome of the seagrass Zostera marina: Do they contribute to pathogen reduction in seawater?

Seagrass meadows provide crucial ecosystem services for coastal environments and were shown to reduce the abundance of waterborne pathogens linked to infections in humans and marine organisms in their vicinity. Among potential drivers, seagrass phenolics released into seawater have been linked to pathogen suppression, but the potential involvement of the seagrass microbiome has not been investigated. We hypothesized that the microbiome of the eelgrass Zostera marina, especially the leaf epiphytes that are at direct interface between the seagrass host and the surrounding seawater, inhibit waterborne pathogens thereby contributing to their removal. Using a culture-dependent approach, we isolated 88 bacteria and fungi associated with the surfaces and inner tissues of the eelgrass leaves (healthy and decaying) and the roots. We assessed the antibiotic activity of microbial extracts against a large panel of common aquatic, human (fecal) and plant pathogens, and mined the metabolome of the most active extracts. The healthy leaf epibiotic bacteria, particularly Streptomyces sp. strain 131, displayed broad-spectrum antibiotic activity superior to some control drugs. Gram-negative bacteria abundant on healthy leaf surfaces, and few endosphere-associated bacteria and fungi also displayed remarkable activities. UPLC-MS/MS-based untargeted metabolomics analyses showed rich specialized metabolite repertoires with low annotation rates, indicating the presence of many undescribed antimicrobials in the extracts. This study contributes to our understanding on microbial and chemical ecology of seagrasses, implying potential involvement of the seagrass microbiome in suppression of pathogens in seawater. Such effect is beneficial for the health of ocean and human, especially in the context of climate change that is expected to exacerbate all infectious diseases. It may also assist future seagrass conservation and management strategies.

Akanthomins A–C, aphidicolin analogs from a fungus Akanthomyces sp., that inhibit cell cycle

Natural products that inhibit cell cycle progression may have potential as anticancer agents. In this study, cell cycle inhibition of microbial culture extracts was screened by fluorescent images using HeLa/Fucci2 cells. The culture extract of a fungus, Akanthomyces sp., inhibited the cell cycle progression at the S/G2/M phases, and bioassay-guided fractionation of the extract afforded three previously undescribed aphidicolin derivatives, namely akanthomins A–C, and an undescribed chromone glycoside, specifically 9-hydroxyeugenetin 9-O-β-d-(4-O-methyl)glucopyranoside, in addition to aphidicolin. The chemical structures of these compounds were elucidated by spectroscopic analysis and chemical derivatization. Using a flow cytometer, akanthomin A and aphidicolin were found to inhibit cell cycle progression at the S phase.