García Pérez, Pascual

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García Pérez, Pascual

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García Pérez, Pascual

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Tecnología de Los Alimentos, Nutrición y Bromatología

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Phenolic profiling and in vitro bioactivities of three medicinal Bryophyllum plants
(Elsevier, 2021-01-14) Ayuso, Manuel; Lozano-Milo, Eva; Pereira, Carla; Dias, María Inés; Calhelha, Ricardo C.; Sokovic, Marina; Ferreira, Isabel C.F.R.; Barros, Lillian; Gallego, Pedro P.; Ivanov, Marija; García Pérez, Pascual; Tecnología de Alimentos, Nutrición y Bromatología
Open Access
Neurofuzzy logic predicts a fine-tuning metabolic reprogramming on elicited Bryophyllum PCSCs guided by salicylic acid
(Frontiers Media, 2022-09-23) Lozano-Milo, Eva; Zhang, Leilei; Landin, Mariana; Lucini, Luigi; Gallego, Pedro P.; Miras Moreno, Begoña; García Pérez, Pascual
Novel approaches to the characterization of medicinal plants as biofactories have lately increased in the field of biotechnology. In this work, a multifaceted approach based on plant tissue culture, metabolomics, and machine learning was applied to decipher and further characterize the biosynthesis of phenolic compounds by eliciting cell suspension cultures from medicinal plants belonging to the Bryophyllum subgenus. The application of untargeted metabolomics provided a total of 460 phenolic compounds. The biosynthesis of 164 of them was significantly modulated by elicitation. The application of neurofuzzy logic as a machine learning tool allowed for deciphering the critical factors involved in the response to elicitation, predicting their influence and interactions on plant cell growth and the biosynthesis of several polyphenols subfamilies. The results indicate that salicylic acid plays a definitive genotype-dependent role in the elicitation of Bryophyllum cell cultures, while methyl jasmonate was revealed as a secondary factor. The knowledge provided by this approach opens a wide perspective on the research of medicinal plants and facilitates their biotechnological exploitation as biofactories in the food, cosmetic and pharmaceutical fields.
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In vitro culture of the endangered plant Eryngium viviparum as dual strategy for its ex situ conservation and source of bioactive compounds
(Springer, 2019-06-15) Moreno Ayuso, Juan Manuel; Ramil‑Rego, Pablo; Gallego, Pedro Pablo; Barreal, M. Esther; García Pérez, Pascual; Tecnología de Alimentos, Nutrición y Bromatología; Facultad de Veterinaria
Different Eryngium species have been used with ornamental, agricultural and medicinal purposes, as a consequence of their chemical constituents. In the southwest Europe the endemic Eryngium viviparum, presents a high risk of extinction and ex situ strategies are high recommended for efficient conservation and re-introduction program. The objective of this study was to satisfy a dual objective: (i) to develop an ex situ conservation strategy through micropropagation and (ii) taking advantage of the extraordinary potential of plant tissue culture, produce a considerable amount of plant material to carry out a preliminary phytochemical study, based on the accumulation of phenolic compounds and their associated antioxidant activity. First a factorial design was conducted in order to study the effect of two cytokinins (6- benzylaminopurine, BAP, and kinetin, KIN), at three levels (0, 1 and 2 mg L−1), on shoot multiplication. Later another factorial design was applied, by using three levels of MS medium salt strength (full, half and quarter- strength) and four sucrose levels (0, 1, 2, and 3%) for improving shoot elongation and rooting. In parallel, a preliminary quantification of total phenolic and flavonoid contents from E. viviparum aerial parts was determined. The simple micropropagation protocol designed allowed obtaining a high rates of shoot multiplication (5.1–5.8 new shoots), rooting (100%) with healthy long roots (3.1–3.5 cm) and plantlet acclimatization (96%). Moderate antioxidant activity was recorded in hydromethanolic extracts from E. viviparum aerial parts. High correlation between total phenolic content and BAP levels in the culture media was found. In conclusion, the micropropagation procedure described here for the endangered E. viviparum can be used as new and very efficient ex situ conservation strategy, and as potential source of antioxidants, conferring an added-value to this plant.
Open Access
Chemometric interpretation of metabolomics signatures: from unsupervised to supervised statistics
(2026-05-21) García Pérez, Pascual; Tecnología de Alimentos, Nutrición y Bromatología
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The metabolomics reveals intraspecies variability of bioactive compounds in elicited suspension cell cultures of three Bryophyllum species
(Elsevier, 2021-02-15) García-Pérez, Pascual; Lucini, Luigi; Gallego, Pedro P.; Miras Moreno, Begoña; García Pérez, Pascual
In this work, the combination of elicited plant suspension cultured cells (PSCCs) with untargeted metabolomics establishes a powerful, cutting-edge strategy to unravel the effects of elicitors on the biosynthetic potential of medicinal plants, as Bryophyllum sp. The PSCC technology constitutes a successful biotechnological system for the study and production of bioactive compounds throughout the inclusion of elicitors with the ability to modulate secondary metabolism. The use of methyl jasmonate (MJ) and salicylic acid (SA) as abiotic elicitors on bryophyllum PSCCs, resulted in differential effects on cell growth and secondary metabolism, depending on the species, including synergistic and antagonistic effects. This fact suggests that both elicitors play a pleiotropic effect on plant secondary metabolism, showing complex interactions, according to the UHPLC-QTOF mass spectrometry profiling. Thus, the combination of both elicitors induced a strong synergistic on B. daigremontianum PSCCs, with 2272 putatively annotated compounds, whereas it caused a negative effect on the secondary metabolism of B. × houghtonii PSCCs, being MJ the only elicitor driving a positive effect, presenting 2972 annotated compounds. Meanwhile, B. tubiflorum PSCCs did not show a significant modulation of secondary metabolism, with 1521 annotated compounds. The metabolite annotation indicated that three families of secondary metabolites were mainly affected by elicitation: phenolic compounds constituted the most affected family by elicitation, mainly represented by flavonoids and lignans; N-containing compounds included glucosinolates, amines, and alkaloids, reported to Bryophyllum sp. for the first time; and terpenoids included mainly phytoalexins and saponins. The results depict a deep genotype-dependent metabolomic reprogramming of secondary metabolism in response to elicitors, thanks to the application of untargeted metabolomics. This knowledge will allow the consideration of Bryophyllum sp. as a valuable source of bioactive compounds, with the potential associated to PSCCs, for being included in food, cosmetic, and pharmaceutical applications.
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Bioactive natural products From the genus Kalanchoe as cancer chemopreventive agents: a review
(2018-11-29) García Pérez, Pascual; M.E. Barreal; Rojo-De Dios, L.; Cameselle-Teijeiro, J.F.; Gallego, P.P.; García Pérez, Pascual; Tecnología de Alimentos, Nutrición y Bromatología; Facultad de Veterinaria
Cancer incidence has increased exponentially worldwide in the last decades despite greater efforts being made in the field of cancer research alongside advances in early-diagnosis techniques. As a multifactorial disease, carcinogenesis is caused by both genetic and epigenetic phenomena, the latter being reversible by living a healthy lifestyle and having a balanced diet. In this sense, the adoption of cancer preventive strategies, known as chemoprevention, emerged as an efficient mechanism to block or suppress cancer initiation and progression. The acceptance of new chemopreventive agents involves long-duration, large preclinical and clinical trials in order to assess a reduction in cancer incidence and avoid eventual toxic side effects. Different phytochemical compounds synthesized by plants have the ability of reverting the deleterious processes associated to carcinogenesis at a cellular level. In fact, common dietary phytochemicals have been widely studied as potential chemopreventive agents due to their beneficial implications on the modulation of inflammation, cell cycle, oxidative stress, cancer cell metabolism, and tumor microenvironments. The use of some species belonging to genus Kalanchoe (subgenus Bryophyllum, Crassulaceae) could be of interest for the development of chemopreventive studies, thanks to their capacity to biosynthesize several bioactive compounds with cytotoxic and other anticancer-associated properties.
Open Access
Adsorption of gallic acid, propyl gallate and polyphenols from Bryophyllum extracts on activated carbon
(Nature Research, 2019-10-15) Losada-Barreiro, Sonia; Gallego, Pedro P.; Bravo-Díaz, Carlos; García Pérez, Pascual; Tecnología de Alimentos, Nutrición y Bromatología
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New strategies for the use of Linum usitatissimum cell factories for the production of bioactive compounds
(Elsevier, 2015-12-18) Almagro Romero, Lorena; Belchí Navarro, Sarai; Sánchez-Pujante, Pedro Joaquín; Pedreño García, María Ángeles; García Pérez, Pascual; Tecnología de Alimentos, Nutrición y Bromatología; Facultad de Veterinaria
In this work, suspension-cultured cells of Linum usitatissimum L. were used to evaluate the effect of two types of cyclodextrins, β-glucan and (Z)-3-hexenol separately or in combination on phytosterol and tocopherol production. Suspension-cultured cells of L. usitatissimum were able to produce high levels of phytosterols in the presence of 50 mM methylated-β-cyclodextrins (1325.96 ± 107.06 μg g dry weight−1) separately or in combination with β-glucan (1278.57 ± 190.10 μg g dry weight−1) or (Z)-3-hexenol (1507.88 ± 173.02 μg g dry weight−1), being cyclodextrins able to increase both the secretion and accumulation of phytosterols in the spent medium, whereas β-glucan and (Z)-3-hexenol themselves only increased its intracellular accumulation. Moreover, the phytosterol values found in the presence of hydroxypropylated-β-cyclodextrins were lower than those found in the presence of methylated-β-cyclodextrins in all cases studied. However, the results showed that the presence of methylated-β-cyclodextrins did not increase the tocopherols production and only an increase in tocopherol levels was observed when cells were elicited with 50 mM hydroxypropylated-β-cyclodextrins in combination with β-glucan (174 μg g dry weight−1) or (Z)-3-hexenol (257 μg g dry weight−1). Since the levels of tocopherol produced in the combined treatment were higher than the sum of the individual treatments, a synergistic effect between both elicitors was assumed. To sum up, flax cell cultures elicited with cyclodextrins alone or in combination with β-glucan or (Z)-3-hexenol were able produce phytosterols and tocopherols, and therefore, these elicited suspension-cultured cells of L. usitatissimum can provide an alternative system, which is at the same time more sustainable, economical and ecological for their production.
Open Access
Machine learning unmasked nutritional imbalances on the medicinal plant Bryophyllum sp. cultured in vitro
(Frontiers Media, 2020-12-01) Lozano-Milo, Eva; Landin, Mariana; Gallego, Pedro Pablo; García Pérez, Pascual; Tecnología de Alimentos, Nutrición y Bromatología
Plant nutrition is a crucial factor that is usually underestimated when designing plant in vitro culture protocols of unexploited plants. As a complex multifactorial process, the study of nutritional imbalances requires the use of time-consuming experimental designs and appropriate statistical and multiple regression analysis for the determination of critical parameters, whose results may be difficult to interpret when the number of variables is large. The use of machine learning (ML) supposes a cutting-edge approach to investigate multifactorial processes, with the aim of detecting non-linear relationships and critical factors affecting a determined response and their concealed interactions. Thus, in this work we applied artificial neural networks coupled to fuzzy logic, known as neurofuzzy logic, to determine the critical factors affecting the mineral nutrition of medicinal plants belonging to Bryophyllum subgenus cultured in vitro. The application of neurofuzzy logic algorithms facilitate the interpretation of the results, as the technology is able to generate useful and understandable “IF-THEN” rules, that provide information about the factor(s) involved in a certain response. In this sense, ammonium, sulfate, molybdenum, copper and sodium were the most important nutrients that explain the variation in the in vitro culture establishment of the medicinal plants in a species-dependent manner. Thus, our results indicate that Bryophyllum spp. display a fine-tuning regulation of mineral nutrition, that was reported for the first time under in vitro conditions. Overall, neurofuzzy model was able to predict and identify masked interactions among such factors, providing a source of knowledge (helpful information) from the experimental data (non-informative per se), in order to make the exploitation and valorization of medicinal plants with high phytochemical potential easier.