In vitro Approaches for Investigating the Bioaccessibility and Bioavailability of Dietary Nutrients and Bioactive Metabolites

The consumption of berry-type fruits has become very popular in recent years because of their positive effects on human health. Berries are in fact widely known for their health-promoting benefits, including prevention of chronic disease, cardiovascular disease and cancer. Berries are a rich source of bioactive metabolites, such as vitamins, minerals, and phenolic compounds, mainly anthocyanins. Numerous in vitro and in vivo studies recognized the health effects of berries and their function as bioactive modulators of various cell functions associated with oxidative stress. Plants have one of the largest metabolome databases, with over 1200 papers on plant metabolomics published only in the last decade. Mass spectrometry (MS) and NMR (Nuclear Magnetic Resonance) are the most important analytical technologies on which the emerging ''omics'' approaches are based. They may provide detection and quantization of thousands of biologically active metabolites from a tissue, working in a ''global'' or ''targeted'' manner, down to ultra-trace levels. In the present review, we highlighted the use of MS and NMR-based strategies and Multivariate Data Analysis for the valorization of berries known for their biological activities, important as food and often used in the preparation of nutraceutical formulations.

Download Full-text

Improved Production of Two Anti-Candida Lipopeptide Homologues Co- Produced by the Wild-Type Bacillus subtilis RLID 12.1 under Optimized Conditions

Current Pharmaceutical Biotechnology ◽

10.2174/1389201020666191205115008 ◽

2020 ◽

Vol 21 (5) ◽

pp. 438-450

Author(s):

Ramya Ramchandran ◽

Swetha Ramesh ◽

Anviksha A ◽

RamLal Thakur ◽

Arunaloke Chakrabarti ◽

...

Keyword(s):

Bacillus Subtilis ◽

Pathogenic Fungi ◽

Fold Increase ◽

Production Yield ◽

Yield Enhancement ◽

Bioactive Metabolites ◽

Candida Auris ◽

Media Formulation ◽

Static Conditions

Background:: Antifungal cyclic lipopeptides, bioactive metabolites produced by many species of the genus Bacillus, are promising alternatives to synthetic fungicides and antibiotics for the biocontrol of human pathogenic fungi. In a previous study, the co- production of five antifungal lipopeptides homologues (designated as AF1, AF2, AF3, AF4 and AF5) by the producer strain Bacillus subtilis RLID 12.1 using unoptimized medium was reported; though the two homologues AF3 and AF5 differed by 14 Da and in fatty acid chain length were found effective in antifungal action, the production/ yield rate of these two lipopeptides determined by High-Performance Liquid Chromatography was less in the unoptimized media. Methods:: In this study, the production/yield enhancement of the two compounds AF3 and AF5 was specifically targeted. Following the statistical optimization (Plackett-Burman and Box-Behnken designs) of media formulation, temperature and growth conditions, the production of AF3 and AF5 was improved by about 25.8- and 7.4-folds, respectively under static conditions. Results:: To boost the production of these two homologous lipopeptides in the optimized media, heat-inactivated Candida albicans cells were used as a supplement resulting in 34- and 14-fold increase of AF3 and AF5, respectively. Four clinical Candida auris isolates had AF3 and AF5 MICs (100 % inhibition) ranging between 4 and 16 μg/ml indicating the lipopeptide’s clinical potential. To determine the in vitro pharmacodynamic potential of AF3 and AF5, time-kill assays were conducted which showed that AF3 (at 4X and 8X concentrations) at 48h exhibited mean log reductions of 2.31 and 3.14 CFU/ml of C. albicans SC 5314, respectively whereas AF5 at 8X concentration showed a mean log reduction of 2.14 CFU/ml. Conclusion:: With the increasing threat of multidrug-resistant yeasts and fungi, these antifungal lipopeptides produced by optimized method promise to aid in the development of novel antifungal that targets disease-causing fungi with improved efficacy.

Download Full-text

Plant-Derived Nano and Microvesicles for Human Health and Therapeutic Potential in Nanomedicine

Pharmaceutics ◽

10.3390/pharmaceutics13040498 ◽

2021 ◽

Vol 13 (4) ◽

pp. 498

Author(s):

Mariaevelina Alfieri ◽

Antonietta Leone ◽

Alfredo Ambrosone

Keyword(s):

Therapeutic Potential ◽

Biological Activities ◽

Plant Biotechnology ◽

Bioactive Metabolites ◽

Anticancer Activities ◽

Long Distance ◽

In Vivo Models ◽

Therapeutic Tools

Plants produce different types of nano and micro-sized vesicles. Observed for the first time in the 60s, plant nano and microvesicles (PDVs) and their biological role have been inexplicably under investigated for a long time. Proteomic and metabolomic approaches revealed that PDVs carry numerous proteins with antifungal and antimicrobial activity, as well as bioactive metabolites with high pharmaceutical interest. PDVs have also been shown to be also involved in the intercellular transfer of small non-coding RNAs such as microRNAs, suggesting fascinating mechanisms of long-distance gene regulation and horizontal transfer of regulatory RNAs and inter-kingdom communications. High loading capacity, intrinsic biological activities, biocompatibility, and easy permeabilization in cell compartments make plant-derived vesicles excellent natural or bioengineered nanotools for biomedical applications. Growing evidence indicates that PDVs may exert anti-inflammatory, anti-oxidant, and anticancer activities in different in vitro and in vivo models. In addition, clinical trials are currently in progress to test the effectiveness of plant EVs in reducing insulin resistance and in preventing side effects of chemotherapy treatments. In this review, we concisely introduce PDVs, discuss shortly their most important biological and physiological roles in plants and provide clues on the use and the bioengineering of plant nano and microvesicles to develop innovative therapeutic tools in nanomedicine, able to encompass the current drawbacks in the delivery systems in nutraceutical and pharmaceutical technology. Finally, we predict that the advent of intense research efforts on PDVs may disclose new frontiers in plant biotechnology applied to nanomedicine.

Download Full-text

Optimization of cultural conditions affecting improved bioactive metabolite production by endophytic fungus Trichoderma harzianum

Bionatura ◽

10.21931/rb/2021.06.04.8 ◽

2021 ◽

Vol 6 (4) ◽

pp. 2187-2192

Author(s):

Rashid Rahim Hateet ◽

Zainab Alag Hassan ◽

Abdulameer Abdullah Al-Mussawi ◽

Shaima Rabeea Banoon

Keyword(s):

Antibacterial Activity ◽

Trichoderma Harzianum ◽

Endophytic Fungus ◽

Nitrogen Sources ◽

Bioactive Metabolites ◽

Bioactive Metabolite ◽

Metabolite Production ◽

Surface Sterilization ◽

Cultural Conditions

The present study aimed to optimize cultural conditions for optimum bioactive metabolite production by endophytic fungus Trichoderma harzianum, isolated by surface sterilization method from the leaf of the eucalyptus plant. The fungus was identified based on morphological characterization. Fungal metabolites were carried out by ethyl acetate solvent. The antibacterial activity was tested against Escherichia coli (ATCC 25922) and Staphylococcus aureus (NCTC 6571). Various carbon, nitrogen sources, pH, temperature, incubation period, and NaCl on the antibacterial metabolite production were studied. Bioactive metabolite production of T. harzianum exhibits a broad spectrum of in vitro antibacterial activity against two strains of bacteria. For the optimum production of bioactive metabolites, Dextrose and Glucose were found to be the best sources of carbon and the best sources of Nitrogen Yeast extract (YE) and (NH4)2SO. The maximum production of bioactive metabolites occurs at pH 7 and 25°C.; the NaCl showed a positive influence on bioactive metabolites.

Download Full-text

Fishing for Targets of Alien Metabolites: A Novel Peroxisome Proliferator-Activated Receptor (PPAR) Agonist from a Marine Pest

Marine Drugs ◽

10.3390/md16110431 ◽

2018 ◽

Vol 16 (11) ◽

pp. 431 ◽

Cited By ~ 12

Author(s):

Rosa Vitale ◽

Enrico D'Aniello ◽

Stefania Gorbi ◽

Andrea Martella ◽

Cristoforo Silvestri ◽

...

Keyword(s):

Native Species ◽

Molecular Mechanisms ◽

Ex Vivo ◽

Invasion Biology ◽

In Vitro Assays ◽

Bioactive Metabolites ◽

Peroxisome Proliferator ◽

Invasive Pests

Although the chemical warfare between invasive and native species has become a central problem in invasion biology, the molecular mechanisms by which bioactive metabolites from invasive pests influence local communities remain poorly characterized. This study demonstrates that the alkaloid caulerpin (CAU)—a bioactive component of the green alga Caulerpa cylindracea that has invaded the entire Mediterranean basin—is an agonist of peroxisome proliferator-activated receptors (PPARs). Our interdisciplinary study started with the in silico prediction of the ligand-protein interaction, which was then validated by in vivo, ex vivo and in vitro assays. On the basis of these results, we candidate CAU as a causal factor of the metabolic and behavioural disorders observed in Diplodus sargus, a native edible fish of high ecological and commercial relevance, feeding on C. cylindracea. Moreover, given the considerable interest in PPAR activators for the treatment of relevant human diseases, our findings are also discussed in terms of a possible nutraceutical/pharmacological valorisation of the invasive algal biomasses, supporting an innovative strategy for conserving biodiversity as an alternative to unrealistic campaigns for the eradication of invasive pests.

Download Full-text

In Vitro Antimicrobial Activity of Piper retrofractum Fruit Extracts against Microbial Pathogens Causing Infections in Human and Animals

International Journal of Microbiology ◽

10.1155/2020/5638961 ◽

2020 ◽

Vol 2020 ◽

pp. 1-6

Author(s):

Wattana Panphut ◽

Tanakwan Budsabun ◽

Pakkakul Sangsuriya

Keyword(s):

Opportunistic Infections ◽

Colorimetric Assay ◽

Disk Diffusion ◽

Bioactive Metabolites ◽

Diffusion Method ◽

Klebsiella Pneumonia ◽

Disk Diffusion Test ◽

Animal Pathogens ◽

Piper Retrofractum

Long pepper (Piper retrofractum Vahl) is a Thai medicinal herb which has been used as one of the common ingredients in variety of Thai foods. Here, we investigated antimicrobial activities of crude bioactive metabolites extracted from fruits of P. retrofractum against 10 pathogenic organisms (bacteria and yeast) causing opportunistic infections in human or animals including Bacillus subtilis ATCC6633, Staphylococcus aureus ATCC25923, Enterococcus faecalis ATCC2921, Escherichia coli ATCC25922, Klebsiella pneumonia TISTR1843, Pseudomonas aeruginosa ATCC741, Salmonella typhi (clinical isolate), Vibrio parahaemolyticus (XN98 and 5HP), and Candida albicans ATCC90020. The results of disk diffusion test showed that the extract from methanol solvent exhibited greater antibacterial activity than other solvents with inhibition zones ranging from 0.5 to 8.0 mm, respectively. Subsequently, minimal inhibition concentration (MIC) determined by the colorimetric assay confirmed that methanol extracts showed consistent results with disk diffusion method. In summary, in vitro assays suggest that methanol is the best solvent for extraction of bioactive metabolites from P. retrofractum fruits. This crude extract can inhibit the majority of human and animal pathogens. This opens up a potential use of pepper fruits in prevention of food-contaminating microorganisms.

Download Full-text

A Metabolomic Study of Epichloë Endophytes for Screening Antifungal Metabolites

Metabolites ◽

10.3390/metabo12010037 ◽

2022 ◽

Vol 12 (1) ◽

pp. 37

Author(s):

Krishni Fernando ◽

Priyanka Reddy ◽

Kathryn M. Guthridge ◽

German C. Spangenberg ◽

Simone J. Rochfort

Keyword(s):

Phytopathogenic Fungi ◽

Bioactive Metabolites ◽

Symbiotic Association ◽

In Planta ◽

Vitro Assay ◽

Antifungal Metabolites ◽

Bioassay Guided Fractionation ◽

Toxic Alkaloids ◽

Epichloë Endophytes

Epichloë endophytes, fungal endosymbionts of Pooidae grasses, are commonly utilized in forage and turf industries because they produce beneficial metabolites that enhance resistance against environmental stressors such as insect feeding and disease caused by phytopathogen infection. In pastoral agriculture, phytopathogenic diseases impact both pasture quality and animal production. Recently, bioactive endophyte strains have been reported to secrete compounds that significantly inhibit the growth of phytopathogenic fungi in vitro. A screen of previously described Epichloë-produced antifeedant and toxic alkaloids determined that the antifungal bioactivity observed is not due to the production of these known metabolites, and so there is a need for methods to identify new bioactive metabolites. The process described here is applicable more generally for the identification of antifungals in new endophytes. This study aims to characterize the fungicidal potential of novel, ‘animal friendly’ Epichloë endophyte strains NEA12 and NEA23 that exhibit strong antifungal activity using an in vitro assay. Bioassay-guided fractionation, followed by metabolite analysis, identified 61 metabolites that, either singly or in combination, are responsible for the observed bioactivity. Analysis of the perennial ryegrass-endophyte symbiota confirmed that NEA12 and NEA23 produce the prospective antifungal metabolites in symbiotic association and thus are candidates for compounds that promote disease resistance in planta. The “known unknown” suite of antifungal metabolites identified in this study are potential biomarkers for the selection of strains that enhance pasture and turf production through better disease control.

Download Full-text

Resolvins as Regulators of the Immune System

The Scientific World JOURNAL ◽

10.1100/tsw.2010.72 ◽

2010 ◽

Vol 10 ◽

pp. 818-831 ◽

Cited By ~ 20

Author(s):

Hiroyuki Seki ◽

Takaharu Sasaki ◽

Tomomi Ueda ◽

Makoto Arita

Keyword(s):

Immune System ◽

Acute Inflammation ◽

Immune Cell ◽

Inflammatory Responses ◽

Bioactive Metabolites ◽

Cell Functions ◽

First Response ◽

Beneficial Impact

Inflammation is the first response of the immune system to infection or injury, but excessive or inappropriate inflammatory responses contribute to a range of acute and chronic human diseases. Clinical assessment of dietary supplementation of ω-3 polyunsaturated fatty acids (i.e., eicosapentaenoic acid [EPA] and docosahexaenoic acid [DHA]) indicate that they have beneficial impact on these diseases, although the mechanisms are poorly understood at the molecular level. In this decade, it has been revealed that EPA and DHA are enzymatically converted to bioactive metabolites in the course of acute inflammation and resolution. These metabolites were shown to regulate immune cell functions and to display potent anti-inflammatory actions bothin vitroandin vivo. Because of their ability to resolve an acute inflammatory response, they are referred to as proresolving mediators, or resolvins. In this review, we provide an overview of the formation and actions of these lipid mediators.

Download Full-text

Biomedical applications of fish gut associated Streptomyces maritimus IM20 isolated from Rastrelliger kanagurta (Indian mackerel)

Research Journal of Biotechnology ◽

10.25303/1610rjbt120130 ◽

2021 ◽

Vol 16 (10) ◽

pp. 120-130

Author(s):

Angamuthu Vignesh ◽

Venugopal Gopikrishnan ◽

Sivaraj Anbarasu ◽

Manikkam Radhakrishnan ◽

Joseph Jerrine

Keyword(s):

Ethyl Acetate ◽

Ethyl Acetate Extract ◽

Radical Scavenging ◽

Bioactive Metabolites ◽

Molecular Characteristics ◽

Inhibition Assay ◽

Indian Mackerel ◽

Fish Gut ◽

Rastrelliger Kanagurta

The biomedical potential of fish gut-associated actinobacteria isolated from the marine fish Rastrelliger kanagurta (Indian mackerel) was investigated. The actinobacterial strain IM20 was isolated from the fish gut by using Kusters agar medium prepared with 50% sea water. Based on their phenotypic and molecular characteristics, strain IM20 was identified as Streptomyces maritimus. The bioactive metabolites produced from the strain IM20 by agar surface fermentation and ethyl acetate extraction were tested for in vitro antimicrobial, antiquorum sensing , anti-biofilm, anti TB, anti-oxidant and anti-cancer activity. The MIC value of ethyl acetate extract (EAE) of IM20 was found to be 16 μg ml−1 against S. aureus and E. coli. In quorum sensing inhibition assay, the extract showed violacein inhibition upto 87% at 512 μg ml−1 concentration when tested by pigment inhibition assay using C. violaceum MTCC 2656. The results of in vitro assays revealed that the ethyl acetate extract of IM-20 (EAE-IM20) showed 75%, 83% and 72% inhibition against Mycobacterium tuberculosis H37Rv, M. tuberculosis (SHRE sensitive) and multi drug resistant (MDR) M tuberculosis respectively at 500 μg ml−1 concentration. In DPPH assay, 71% radical scavenging activity was exhibited by the EAE-IM20 at 250 μg ml−1. In MTT assay, EAEIM20 exhibited 74.29±2.01% inhibition on breast cancer cell line MCF7 (250 μg ml−1). This study broadly determines that the fish associated actinobacteria is a prolific place for diverse multifunctional bioactive compounds for the development of medically important unique drugs.

Download Full-text

Monascin and monascinol, azaphilonoid pigments from Mortierella polycephala AM1: in silico and in vitro targeting of the angiogenic VEGFR2 kinase

Zeitschrift für Naturforschung C ◽

10.1515/znc-2021-0095 ◽

2021 ◽

Vol 0 (0) ◽

Author(s):

Mohamed Shaaban ◽

Mohammad Magdy El-Metwally ◽

Amal A. I. Mekawey ◽

Ahmed B. Abdelwahab ◽

Maha M. Soltan

Keyword(s):

In Silico ◽

Topoisomerase Ii ◽

Potent Inhibitor ◽

Bioactive Metabolites ◽

Cytotoxic Activities ◽

Feather Waste ◽

Biological Target ◽

Topo Ii ◽

Poultry Feather

Abstract The fungus, Mortierella polycephala is one of the most productive sources of anticancer bioactive compounds namely those of pigment nature. During our investigation of the produced bioactive metabolites by the terrestrial M. polycephala AM1 isolated from Egyptian poultry feather waste, two main azaphilonoid pigments, monascin (1) and monascinol (2) were obtained as major products; their structures were identified by 1D (1H&13C) and 2D (1H–1H COSY, HMBC) NMR and HRESI-MS spectroscopic data. Biologically, cytotoxic activities of these compounds were broadly studied compared with the fungal extract. To predict the biological target for the presumed antitumor activity, an in silico study was run toward three proteins, topoisomerase IIα, topoisomerase IIβ, and VEGFR2 kinase. Monascinol (2) was expected to be moderately active against VEGFR2 kinase without any anticipated inhibition toward topo II isoforms. The in vitro study confirmed the docked investigation consistently and introduced monascinol (2) rather than its counterpart (1) as a potent inhibitor to the tested VEGFR2 kinase. Taxonomically, the fungus was identified using morphological and genetic assessments.

Download Full-text