Unravelling the biotechnological potential of halophyte species from the Algarve coast - HaloBiotech

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If you cannot beat them, join them: exploring the fruits of the invasive species Carpobrotus edulis (L.) NE Br as a source of bioactive products

Publication . Castañeda-Loaiza, Viana; Placines, Chloé; Rodrigues, Maria João; Pereira, Catarina; Zengin, Gokhan; Uysal, Ahmet; Jeko, Jozsef; Cziaky, Zoltan; Reis, Catarina Pinto; Gaspar, Maria Manuela; Custódio, Luísa

The halophyte species Carpobrotus edulis (L.) N.E. Br, also known as Hottentot-fig, is one of the 20 most aggressive invasive species of coastal areas worldwide. It is native to South Africa, where it is used in traditional medicine for the treatment of several diseases, including tuberculosis and acquired immunodeficiency syndrome (AIDS) caused by the human immunodeficiency virus (HIV). Aiming at a sustainable use of its biomass as a value-added product, this work reports for the first time the in vitro antioxidant, anti-microbial, enzymatic inhibitory properties and toxicity of peel and flesh extracts of Hottentot-fig mature fruits. The extracts' chemical composition was also determined by spectrophotometric methods (total contents of phenolics: TPC; flavonoids: TFC and tannins: TTC), and by high-performance liquid chromatography coupled with electrospray ionization mass spectrometry (HPLC-ESI-MS/MS). The peels' extracts had generally the highest TPC, TFC and TTC, especially the ethanol ones (TPC: 272.82 mg gallic acid equivalents (GAE)/g dry weight (DW), TFC: 1.58 mg quercetin equivalents (QE)/g DW and TTC: 20.3 mg catechin equivalents (CE)/g DW). The peels' extracts also had the highest diversity of compounds, mostly phenolic acids, flavonoids, and coumarins, as identified by HPLC-ESI-MS/MS. Some molecules were specific to a particular fruit part, for example, coumaric acid and uvaol in the peel, and vanillin and kaempferol-O-(rhamnosyl)hexosylhexoside in the flesh. Some compounds are here described for the first time in Hottentot-fig, such as azelaic acid and emodin. The peel's extracts had the highest anti radical activity, especially the ethanol and acetone towards 2,2-diphenyl-1-picrylhydrazyl (DPPH) (half maximal inhibitory concentration (IC50) values of 0.59 and 0.88 mg/mL, respectively), and the acetone extract against 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) (IC50 = 0.56 mg/mL). Samples had nil capacity to chelate iron, a low copper chelation potential, but a significant capacity to reduce iron, especially the ethanol (IC50 = 0.09 mg/mL) and the acetone extracts of peels (IC50 = 0.10 mg/mL) and flesh (IC50 = 0.11 mg/mL) and also the water peel's extracts (IC50 = 0.18 mg/mL). Samples had nil to low activity towards the enzymes acetylcholinesterase (AChE), butyrylcholinesterase (BuChE), alpha-amylase and alpha-glucosidase, but displayed a strong inhibition of tyrosinase, especially the ethanol peel's extracts (29.55 mg kojic acid equivalents (KAE)/g). Samples had nil to low in vitro toxicity towards human keratinocytes. All together our results suggests possible novel biotechnological applications of Hottentot-fig fruits as sources of innovative bioactive ingredients for the food, cosmetic, agriculture and/or pharmaceutical industries.

2020-02Journal article

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In vitro and in silico approaches to appraise Polygonum maritimum L. as a source of innovative products with anti-ageing potential

Publication . Rodrigues, Maria Joao; Slusarczyk, Sylwester; Pecio, Łukasz; Matkowski, Adam; Salmas, Ramin Ekhteiari; Durdagi, Serdar; Pereira, Catarina; Varela, J.; Barreira, Luísa; Custódio, Luísa

Different Polygonum species have in vitro neuroprotective properties and are traditionally used for their anti-ageing benefits. In this context this work explored for the first time P. maritimum (sea knotgrass) as a potential source of natural products with industrial application as cognitive enhancers with anti-ageing potential. For that purpose methanol and dichloromethane extracts were prepared from leaves and roots of that species and evaluated for in vitro neuroprotective effects through the inhibition on acetyl- (AChE) and butyrylcholinesterase (BuChE), protection against hydrogen peroxide (H2O2)-induced cytoxicity on neuroblastoma cells and lipopolysaccharide (LPS)-induced neuroinflammation on microglia cells. The extracts were chemically characterized by ultra-high-pressure liquid chromatography (UHPLC) coupled to tandem mass spectrometry (MS/MS) and docking studies were performed on the identified compounds. Methanol extracts had the highest activity in AChE (leaves: IC50 = 0.27 mg/mL; roots: IC50 = 0.17 mg/mL) and BuChE (leaves: IC50 = 0.62 mg/mL; roots: IC50 = 0.61 mg/mL) inhibition, as well as reduction of nitric oxide (NO) production in LPS-treated microglia (leaves: IC50 = 4.17 μg/mL; roots: IC50 = 9.95 μg/mL). Methanol extracts prevented oxidative stress-induced cytotoxicity in SH-SY5Y cells when applied simultaneously with H2O2, whereas cells pre-treated with the dichloromethane extracts had increased viability. Fifteen flavonoids were identified and showed favorable binding energies to AChE and BuChE binding pockets. These data suggests P. maritimum as a promising source of natural products and/or molecules with cognitive enhancement and anti-ageing properties.

2018-01Journal article

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Synchronous insight of in vitro and in vivo biological activities of Sambucus nigra L. extracts for industrial uses

Publication . Mota, Ana Henriques; Andrade, Joana M.; Rodrigues, Maria Joao; Custódio, Luísa; Bronze, Maria Rosario; Duarte, Noelia; Baby, Andre; Rocha, Joao; Gaspar, Maria Manuela; Simoes, Sandra; Carvalheiro, Manuela; Fattal, Elias; Almeida, Antonio Jose; Reis, Catarina Pinto

There is a re-emerging interest in natural products as reputable sources of new active pharmaceutical ingredients. This study synchronously reports in vitro, with more than one cell line, and in vivo biological activities of extracts obtained from Sambucus nigra. Using several solvents and techniques, eighteen extracts were obtained from fresh and dried berries, and fresh flowers. The flavonoid content and identification were determined using HPLC-MS/MS. The extracts were then screened for antioxidant activity, total polyphenol content, collagenase, elastase, tyrosinase and acetylcholinesterase inhibition as well as photoprotection. In vitro and in vivo (murine model) anti-inflammatory activity and cytotoxicity (skin and monocytic cells) were also studied. The most promising extracts were those obtained from fresh flowers using either ultrasounds or methanol. These extracts showed similar results to positive controls, particularly the antioxidant activity (74.5 +/- 1.6 %), collagenase inhibition (93.6 +/- 0.6 %), photoprotection (Sun Protection Factor > 50), in vitro anti-inflammatory activity (96.9 +/- 2.9 %), as well as oral/topical anti-inflammatory activity. The ultrasounds/ethanol extract of fresh flowers presented higher collagenase inhibition (88.3 +/- 2.8 %) and in vitro anti-inflammatory activity (101.8 +/- 1.5 %). Cytotoxicity testing confirmed the safety. Chemical characterization allowed the deduction of a correlation between extract composition and biological activities, suggesting a straightforward application in the development of novel products subject to further investigation.

2020-10Journal article

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Exploring Ulva australis Areschoug for possible biotechnological applications: In vitro antioxidant and enzymatic inhibitory properties, and fatty acids contents

Publication . Trentin, Riccardo; Custódio, Luísa; Rodrigues, Maria Joao; Moschin, Emanuela; Sciuto, Katia; Da Silva, José Paulo; Moro, Isabella

Ulva australis Areschoug (Ulvales, Chlorophyta) is traditionally used as a foodstuff in Korea and to treat several diseases, such as hyperlipidemia and urinary ailments. It is now considered an invasive species of the Venice Lagoon (Italy). This work explored U. australis as a potential source of bioactive molecules with biotechnological applications. For that purpose, organic and water extracts were prepared from dried biomass of U. australis and evaluated for radical scavenging activity (RSA), metal chelating activities and for in vitro inhibition of enzymes related with several disorders, namely Alzheimer's disease (acetylcholinesterase, butyrylcholinesterase), hyperpigmentation (tyrosinase), obesity (lipase), Type-2 diabetes mellitus (alpha-glucosidase, alpha-amylase) and skin sagging (elastase). Extracts were also evaluated for total contents of phenolics, flavonoids condensed tannins and carbohydrates. Finally, the fatty acids methyl esters profile of the biomass was established by gas chromatography-mass spectrometry. Extracts had low to moderate radical scavenging activity on DPPH, and a higher ability to chelate Fe2+ than Cu2+, and the maximum Fe2+ chelating capacity was observed in the tincture (IC50 = 4.82 mg/mL). Samples had a higher ability to inhibit alpha-amylase and microbial alpha-glucosidase than the other tested enzymes. The biomass of U. australis presented a highly saturated fatty acids profile, enriched in palmitic acid. These data suggest U. australis as a promising source of natural products with anti- diabetic properties and as a desirable species for biodiesel production.

2020-09Journal article

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