Extraction and analysis of plant volatiles

Mestrado de dupla diplomação com a Université Libre de TunisAs worldwide population is growing, the demand on food increases exponentially, which results in food overproduction, food spoilage, and in foodborne diseases. This issue has arisen many concerns related to public health, economy, and the environment, which led to exhaustive research on possible solutions to minimize costs and ensure safer food products that respond to consumers’ demand for clean-label foods with less collateral damages on the environment. Essential oils (E.Os) and volatiles are plant secondary metabolites and are considered as promising food preservatives seen their remarkable biological activities including antioxidant, antimicrobial, antiproliferative, and anti-inflammatory activities. E.Os from three aromatic plants: eucalyptus (Eucalyptus globulus Labill.), peppermint (Mentha x piperita L.), and pine (Pinus pinaster) were extracted by hydro distillation (HD) and microwave-assisted hydro distillation (MAHD), both optimized with response surface methodology (RSM), which generally showed higher yields in E.Os obtained with MAHD influenced primarily by solid/liquid ratio, with eucalyptus giving the highest yield from the three studied plants. Chromatographic identification resulted in high amounts of monoterpenes, followed by sesquiterpenes in both eucalyptus and peppermint E.Os, while pine E.O was proven to be richer in diterpenes. These volatile compounds led to interesting bioactive properties in all of the studied E.Os, especially the ones extracted using hydro distillation. The extracts showed an inhibitory capacity against the majority of the tested bacterial strains, specifically against Methicillin-resistant Staphylococcus aureus (MRSA), which is a multi- resistant bacteria. Also, a remarkable antifungal activity against Aspergillus brasiliensis and Aspergillus fumigatus. In terms of antiproliferative activity, the E.Os extracted through HD demonstrated higher tumoral cell growth inhibition against the majority of the studied cancer cell lines, with pine E.O standing out with the lowest GI50 value, and none of the tested E.O showed cytotoxicity against normal cell line studied in this case (PLP2). The E.Os also revealed an anti-inflammatory capacity that stood out mainly with those extracted using HD. This work highlights the promising use of essential oils as headspace preservatives for food packaging.Com o crescimento da população mundial, a necessidade de alimentos aumenta exponencialmente, o que resulta em sobreprodução de alimentos, desperdícios e doenças transmitidas por alimentos. Esta questão tem despertado muitas preocupações relacionadas com a saúde pública, economia e meio ambiente, o que levou a investigação sobre possíveis soluções para minimizar custos e garantir produtos alimentares mais seguros e que atendam à procura dos consumidores por alimentos sustentáveis e com menos danos ao ambiente. Os óleos essenciais (O.Es) e moléculas voláteis são metabolitos secundários de plantas e são considerados conservantes de alimentos promissores devido às suas notáveis atividades biológicas, incluindo atividades antioxidantes, antimicrobianas, antiproliverativas, e anti- inflamatórias O.Es de três plantas aromáticas: eucalipto (Eucalyptus globulus Labill.), hortelã-pimenta (Mentha x piperita L.) e pinheiro (Pinus pinaster L. ) foram extraídos por hidrodestilação (HD) e hidrodestilação assistida por microondas (MAHD), ambas otimizada com a metodologia de superfície de resposta (RSM), que apresentou geralmente maiores rendimentos em O.Es. obtidos com MAHD influenciados principalmente pela relação sólido/líquido, onde o eucalipto apresentou o maior rendimento das três plantas estudadas. A identificação cromatográfica resultou em altas quantidades de monoterpenos, seguidos de sesquiterpenos em O.Es de eucalipto e hortelã-pimenta, enquanto o O.E. de pinheiro provou ser mais rico em diterpenos. Esses compostos voláteis levaram a interessantes propriedades bioativas em todos os E.Os estudados, especialmente os extraídos por hidrodestilação. Os extratos mostraram capacidade inibitória contra a maioria das bacterias testadas, especificamente contra Staphylococcus aureus resistente à meticilina (MRSA), que é uma bactéria multirresistente. Além disso, uma notável atividade antifúngica foi detetada contra Aspergillus brasiliensis e Aspergillus fumigatus. Em termos de atividade antiproliferativa, os O.Es extraídos por HD demonstraram maior inibição do crescimento de células tumorais contra a maioria das linhagens de células cancerígenas estudadas, destacando-se o O.E. de pinheiro com o menor valor de GI50, e nenhum dos O.E. testados apresentou citotoxicidade contra a linha não tumoral estudada (PLP2). Os O.E. também revelaram uma capacidade anti-inflamatória que se destacou principalmente com os extraídos com HD. Este estudo revela uma boa possibilidade dos óleos essenciais poderem vir a ser estudados como conservantes alimentares em embalagens alimentares

Non-alkaloid nitrogen-containing compounds from fungi

Fungi represent an ancient ubiquitous and interesting kingdom that can be classified into Phycomycota, Oomycota, Microsporidiomycota, Zygomycota, Ascomycota, Basidiomycota, and Deuteromycota divisions. These microorganisms are considered as a reservoir of bioactive compounds that can be exploited in crucial fields: food, pharmaceutical, and agricultural industries. Non-alkaloid nitrogenated compounds are considered a prolific bouquet composed of proteins such as enzymes playing an essential role in the synthesis of compounds that are involved in vital life processes, lytic enzymes possessing depleting effects, antibiotics, toxins applied as antimicrobials and antifungals, immunosuppressive molecules, etc. Moreover, a number of species found in phyla like Oomycota and Microsporidiomycota have been and are still used as biological control agents for crops as a counterpart to chemical products.The research leading to these results was supported by MICINN supporting the Ramón y Cajal grant for M.A. Prieto (RYC-2017-22891), the post-doctoral grant of M. Fraga- Corral (ED481B-2019/096), and L. Cassani (ED481B-2021/152), and the pre-doctoral grant of A.G. Pereira (ED481A-2019/0228). Authors are grateful to Ibero-American Program on Science and Technology (CYTED-AQUA-CIBUS, P317RT0003), to the Bio Based Industries Joint Undertaking (JU) under grant agreement No 888003 UP4HEALTH Project (H2020-BBI-JTI-2019). The JU receives support from the European Union’s Horizon 2020 research and innovation program and the Bio Based Industries Consortium. The project SYSTEMIC Knowledge hub on Nutrition and Food Security, has received funding from national research funding parties in Belgium (FWO), France (INRA), Germany (BLE), Italy (MIPAAF), Latvia (IZM), Norway (RCN), Portugal (FCT), and Spain (AEI) in a joint action of JPI HDHL, JPI-OCEANS and FACCE-JPI launched in 2019 under the ERA-NET ERA-HDHL (n° 696295). The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to the CIMO (UIDB/00690/2020). I. Oliveira thanks FCT for her PhD grant (BD/06017/2020). M. Carocho thanks FCT for his individual scientific contract (CEEC-IND/00831/2018).info:eu-repo/semantics/publishedVersio