Browsing by Subject "Cyclodextrins"
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- PublicationEmbargoA rational approach to improving the biotechnological production of taxanes in plant cell cultures of Taxus spp.(Elsevier, 2014-03-27) Cusido, Rosa M.; Onrubia, Miriam; Sabater Jara, Ana Belén; Moyano, Elisabeth; Bonfill, Mercedes; Goossens, Alain; Pedreño, María Angeles; Palazón, Javier; Biología VegetalTaxol is a complex diterpene alkaloid scarcely produced in nature and with a high anticancer activity. Biotechnological systems for taxol production based on cell cultures of Taxus spp. have been developed, but the growing commercial demand for taxol and its precursors requires the optimization of these procedures. In order to increase the biotechnological production of taxol and related taxanes in Taxus spp. cell cultures, it is necessary not only to take an empirical approach that strives to optimize in-put factors (cell line selection, culture conditions, elicitation, up-scaling, etc.) and out-put factors (growth, production, yields, etc.), but also to carry out molecular biological studies. The latter can provide valuable insight into how the enhancement of taxane biosynthesis and accumulation affects metabolic profiles and gene expression in Taxus spp. cell cultures. Several rational approaches have focused on studying the transcriptomic profiles of key genes in the taxol biosynthetic pathway in Taxus spp. cell cultures treated with elicitors such as methyl jasmonate, coronatine and cyclodextrins in relation with the taxane pattern, production and excretion to the culture medium. These studies have provided new insights into the taxol biosynthetic pathway and its regulation. Additionally, identifying genes with low levels of expression even in the presence of elicitors, together with metabolomics studies, has shed light on the limiting steps in taxol biosynthesis and could help define suitable metabolic targets for engineering with the main aim of obtaining highly productive Taxus cultured cells. In this review, we have summarized the latest endeavors to enhance the molecular understanding of the action mechanism of elicitors in Taxus spp. cell cultures. Developments in the ongoing search for new and more effective elicitation treatments and the application of metabolic engineering to design new transgenic cell lines of Taxus with an improved capacity for taxane production are described.
- PublicationOpen AccessA way to increase the bioaccesibility and photostability of roflumilast, a COPD treatment, by cyclodextrin monomers(MDPI, 2019-04-30) Matencio, Adrián; Hernández García, Samanta; García Carmona, Francisco; López Nicolás, José Manuel; Bioquímica y Biología Molecular ARoflumilast is an orally available inhibitor of phosphodiesterase (PDE) type 4, which is widely used in chronic obstructive pulmonary diseases. However, it has low solubility and adverse effects include diarrhea and nausea. Since its solubilization may improve treatment and, dismissing any adverse effects, its interaction with cyclodextrins (CDs) was studied. The Higuchi-Connors method was used to determine the complexation constant with different CDs, pH values and temperatures. Molecular docking was used to predict interaction between the complexes. An in vitro digestion experiment was carried out to test roflumilast protection. Finally, the photostability of the complex was evaluated. The complex formed with β-CD had the highest K11 value (646 ± 34 M−1), although this value decreased with increasing temperature. Similarly, K11 decreased as the pH increased. In vitro digestion showed that CDs protect the drug during digestion and even improve its bioaccessibility. Finally, CDs reduced the drug’s extreme photosensitivity, originating a fluorescence signal, which is described for first time. The kinetic parameters of the reaction were obtained. This study not only completes the complexation study of roflumilast-CD, but also points to the need to protect roflumilast from light, suggesting that tablets containing the drug might be reformulated.
- PublicationRestrictedApplications of cyclodextrins in food science. A Review(Elsevier, 2020-08-25) Matencio, Adrián; Navarro Orcajada, Silvia; García Carmona, Francisco; López Nicolás, José Manuel; Bioquímica y Biología Molecular ABackground: The food industry is constantly attempting to develop better products that will have a positive effect on health (commonly known as functional foods). In this respect, cyclodextrins (CDs) could be of interest because they are tasteless, non-caloric and odourless molecules with several valuable characteristics, such as a capacity to separate chiral compounds and solubilize or stabilize bioactive compounds (BaC). Scope and approach: This review represents a revision of the state-of-the-art of CDs and their uses in the food industry. Key findings and conclusions: We analysed their metabolism and regulatory aspects of current applications of CDs: as carriers, for removing components, to produce or extract BaC, their use as nanosensors or in food packaging. We study how inclusion complexed are formed referring to the most common techniques and parameters Moreover, how inclusion complexes are formed will be studied with reference to the most common techniques and parameters. In conclusion, their applications in the food and other industries will increase in the coming years without a doubt.
- PublicationEmbargoApplications of cyclodextrins in food science. A review(Elsevier, 2020-08-25) Matencio, Adrián; Navarro Orcajada, Silvia; García Carmona, Francisco; López Nicolás, José Manuel; Bioquímica y Biología Molecular ABackground: The food industry is constantly attempting to develop better products that will have a positive effect on health (commonly known as functional foods). In this respect, cyclodextrins (CDs) could be of interest because they are tasteless, non-caloric and odourless molecules with several valuable characteristics, such as a capacity to separate chiral compounds and solubilize or stabilize bioactive compounds (BaC). Scope and approach: This review represents a revision of the state-of-the-art of CDs and their uses in the food industry. Key findings and conclusions: We analysed their metabolism and regulatory aspects of current applications of CDs: as carriers, for removing components, to produce or extract BaC, their use as nanosensors or in food packaging. We study how inclusion complexed are formed referring to the most common techniques and parameters Moreover, how inclusion complexes are formed will be studied with reference to the most common techniques and parameters. In conclusion, their applications in the food and other industries will increase in the coming years without a doubt.
- PublicationOpen AccessBiotechnological approach to increase oxyresveratrol production in mulberry in vitro plants under elicitation(MDPI, 2023-01-25) Sabater Jara, Ana Belén; Almagro, Lorena; Nicolás Sánchez, Isabel; Pedreño, María Angeles; Biología VegetalMorus alba L. is used for a range of therapeutic purposes in Asian traditional medicine, and its extracts are reported to be effective against lipidemia, diabetes, and obesity, as well as being hepatoprotective and tyrosinase-inhibitory. They are also included in cosmetic products as anti-aging and skin-whitening agents. Stilbenes, the major bioactive compounds found in M. alba, have received renewed attention recently because of their putative activity against COVID-19. In this study M. alba plants were established in vitro, and the effect of elicitation on plant growth and stilbene accumulation, specifically oxyresveratrol and trans-resveratrol, was investigated. Different concentrations of the elicitors including methyl jasmonate and cyclodextrins were applied, and stilbene levels were determined in leaves, roots, and the culture medium. Elicitation of the M. alba plants with 5 mM cyclodextrins, alone or in combination with 10 µM methyl jasmonate, significantly increased the total phenolic content in the culture medium and leaves after 7 days of treatment. The higher total phenolic content in the roots of control plants and those treated only with methyl jasmonate indicated that cyclodextrins promoted metabolite release to the culture medium. Notably, the cyclodextrin-treated plants with the highest levels of oxy- and trans-resveratrol also had the highest total phenolic content and antioxidant capacity. These results indicate that elicited M. alba in vitro plants constitute a promising alternative source of bioactive stilbenes to supply pharmaceutical and cosmeceutical industries.
- PublicationOpen AccessCyclodextrins increase triterpene production in Solanum lycopersicum cell cultures by activating biosynthetic genes(MDPI, 2022-10-20) Sabater Jara, Ana Belén; Marín Marín, María Jesús; Almagro, Lorena; Pedreño, María Angeles; Biología VegetalIn this work, Solanum lycopersicum cv. Micro-Tom suspension-cultured cells were used to analyze the effect of different elicitors including β-cyclodextrins (CD), methyl jasmonate (MJ), β-glucan (Glu) and 3-hexenol (Hex) separately and the combined treatments of CD + MJ, CD + glu and CD + Hex on triterpene compound production after 24, 72 and 96 h. Moreover, we studied the changes induced by elicitors in the expression of key biosynthetic genes to elucidate the regulation of the triterpene biosynthetic pathway. The relative abundance of the triterpene compounds identified in the extracellular medium after elicitation (squalene, fucosterol, avenasterol, β-sitosterol, cycloartenol and taraxasterol) was determined by gas chromatography coupled to mass spectrometry, and the expression level of genes in treated-cells was analyzed by real-time quantitative polymerase chain reaction (qRT-PCR). Results showed that, in CD-treated cells (CD, CD + MJ, CD + Glu, CD + Hex), specialized metabolites were accumulated mainly in the extracellular medium after 72 h of elicitation. Moreover, qRT-PCR analysis revealed that the highest triterpene levels in CD-treated cells (CD, CD + MJ, CD + Glu, CD + Hex) were highly correlated with the expression of cycloartenol synthase, 3-hydroxy-3-methylglutaryl-CoA reductase and squalene epoxidase genes at 24 h of treatment, whereas the expression of sterol methyltransferase was increased at 72 h. According to our findings, CD acts as a true elicitor of triterpene biosynthesis and can promote the release of bioactive compounds from the tomato cells into the extracellular medium. The results obtained provide new insights into the regulation of the triterpene metabolic pathway, which might be useful for implementing metabolic engineering techniques in tomato.
- PublicationEmbargoEffect of terbinafine on the biosynthetic pathway of isoprenoid compounds in carrot suspension cultured cells(Springer, 2018-04-21) Miras Moreno, Begoña; Almagro, Lorena; Pedreño, María Angeles; Sabater Jara, Ana Belén; Biología VegetalPlant sterols are essential components of membrane lipids, which contributing to their fluidity and permeability. Besides their cholesterol-lowering properties, they also have anti-inflammatory, antidiabetic and anticancer activities. Squalene, which is phytosterol precursor, is widely used in medicine, foods and cosmetics due to its anti-tumor, antioxidant and anti-aging activities. Nowadays, vegetable oils constitute the main sources of phytosterols and squalene, but their isolation and purification involve complex extraction protocols and high costs. In this work, Daucus carota cell cultures were used to evaluate the effect of cyclodextrins and terbinafine on the production and accumulation of squalene and phytosterols as well as the expression levels of squalene synthase and cycloartenol synthase genes. D. carota cell cultures were able to produce high levels of extracellular being phytosterols in the presence of cyclodextrins (12 mg/L), these compounds able to increase both the secretion and accumulation of phytosterols in the culture medium. Moreover, terbinafine induced a significant increase in intracellular squalene production, as seen after 168 h of treatment (497.0 ± 23.5 µg g dry weight−1) while its extracellular production only increased in the presence of cyclodextrins.The analysis of sqs and cas gene expression revealed that cyclodextrins did not induce genes encoding enzymes involved in the phytosterol biosynthetic pathway since the expression levels of sqs and cas genes in cyclodextrin-treated cells were lower than in control cells. The results, therefore, suggest that cyclodextrins were only able to release phytosterols from the cells to the extracellular medium, thus contributing to their acumulation. To sum up, D. carota cell cultures treated with cyclodextrins or terbinafine were able to produce high levels of phytosterols and squalene, respectively, and, therefore, these suspension-cultured cells of carrot constitute an alternative biotechnological system, which is at the same time more sustainable, economic and ecological for the production of these bioactive compounds.
- PublicationEmbargoEnhanced accumulation of phytosterols and phenolic compounds incyclodextrin-elicited cell suspension culture of Daucus carota(Elsevier, 2016-06-15) Miras Moreno, Begoña; Almagro Romero, Lorena; Pedreño, María Angeles; Sabater Jara, Ana Belén; Biología VegetalInthis work, suspension-culturedcells of Daucus carota wereusedto evaluate the effect of -cyclodextrins on the production of isoprenoid and phenolic compounds. The results showed that the phytosterols and phenolic compounds were accumulated in the extracellular medium (15100 g L−1 and 477.46 g L−1, respectively) in the presence of cyclodextrins. Unlike the phytosterol and phenolic compound content, -carotene (1138.03 g L−1), lutein (25949.54 g L−1) and -tocopherol (8063.82 g L−1) chlorophyll a (1625.13 g L−1) and b (9.958 (9958.33 g L−1) were mainly accumulated inside the cells. Therefore, cyclodextrins were able to induce the cytosolic mevalonate pathway, increasing the biosynthesis of phytosterols and phenolic compounds, and accumulate them outside the cells. However, in the absence of these cyclic oligosaccharidic elicitors, carrot cells mainly accumulated carotenoids through the methylerythritol 4-phosphate pathway. Therefore, the use of cyclodextrins would allow the extracellular accumulation of both phytosterols andphenolic compounds bydiverting the carbonflux towards the cytosolicmevalonate/phenylpropanoid pathway.
- PublicationEmbargoEvaluation of juice and milk “food models” fortified with oxyresveratrol and β-Cyclodextrin(Elsevier, 2019-07-25) Matencio, Adrián; Navarro Orcajada, Silvia; Conesa, Irene; Muñoz Sánchez, Iván; Laveda Cano, Lorena; Cano Yelo, Desiré; García Carmona, Francisco; López Nicolás, José Manuel; Bioquímica y Biología Molecular ADespite being saturated, a huge quantity of “novel” functional food products reaches the food market every day and companies are looking for new products to catch consumers’ attention. In the present study, the inclusion complexes of oxyresveratrol with α-, β- and γ-CD are characterized using DSC, TGA, SEM and molecular docking and their complexing capacities arecompared. As β-CD showed the best results, i) 0.2mM oxyresveratrol alone, ii) 0.2mM oxyresveratrol complexed with 8mM β-CD and iii) 4mM oxyresveratrol solubilized with 8mM β-CD were used to fortify, juice and milk food models, and kept in typical storage conditions (darkness and/or refrigerated) conditions for one month. The results showed that CD supplementation leads to a higher oxyresveratrol concentration and antioxidant capacity than when not used. Oxyresveratrol/β-CD food models were stable for five weeks. The most typical variables measured were compared. In vitro digestion pointed to similar bioaccessibility. The bacteriostatic effect was also studied and the greater effectiveness of oversaturated solutions was demostrated. These results should be useful for the food industry for designing hydrophilic products containing oxyresveratrol.
- PublicationEmbargoProduction and localization of hydrogen peroxide and nitric oxide in grapevine cells elicited with cyclodextrins and methyl jasmonate(Elsevier, 2019-06) Belchi Navarro, Sarai; Abellán Rubio, Marina; Pedreño, María Ángeles; Almagro Romero, Lorena; Biología VegetalThe use of methyl jasmonate, alone or in combination with cyclic oligosaccharides such as cyclodextrins, has proved to be a successful strategy for increasing the production of trans-resveratrol in Vitis vinifera cell cultures. However, understanding the intracellular signalling pathways involved in its production would improve the management of grapevine cells as biofactories of this high-value natural product. The results obtained herein confirm the involvement of hydrogen peroxide and nitric oxide in cyclodextrins and methyl jasmonate-induced trans-resveratrol production in grapevine cell cultures. In fact, methyl jasmonate led to maximal intracellular levels of hydrogen peroxide and nitric oxide after 24 h of treatment, but extracellular hydrogen peroxide was only detected in the culture medium when grapevine cells were treated with cyclodextrins. The results derived from the cytochemical detection of H2O2 in elicited grapevine cell cultures also suggested that the combined treatment with cyclodextrins and methyl jasmonate not only increased the production of H2O2 but also released cell wall fragments with electron-dense deposits. Moreover, nitric oxide was localized in all the cellular compartments, particularly in the nucleus and cyto-plasmic organelles, whereas hydrogen peroxide was mainly found in cytoplasmic areas close to the cell wall, and in the nucleoplasm.
- PublicationEmbargoStudy of oxyresveratrol complexes with insoluble cyclodextrin based nanosponges: Developing a novel way to obtain their complexation constants and application in an anticancer study(Elsevier, 2019-12-17) Matencio, Adrián; Kumar Dhakarb, Nilesh; Bessonec, Federica; Musso, Giorgia; Cavallic, Roberta; Dianzanic, Chiara; García Carmona, Francisco; López Nicolás, José Manuel; Bioquímica y Biología Molecular AThe complexation of the bioactive compound oxyresveratrol (OXY) with a polymer called cyclodextrin-based nanosponge (CD-NS) and its application was studied.A new methodology is used to calculate, an apparent inclusion complex constant (KFapp) between a ligand and CD-NSs. Moreover, the KFapp of resveratrol was also evaluated and compared. The complex of OXY with the nanosponge β-CDI 1:4, was studied in vitro using DSC, TGA and FTIR techniques and its drug loading and release behavior were studied. An in vitro digestion showed higher protection of OXY complexed than free OXY. The bioactivity enhancing capacity of OXY was also studied against prostate (PC-3) and colon (HT-29 and HCT-116) cancer cell lines, where it showed stronger cell viability inhibition than the free drug. The findings as a whole represent a new opportunity for studying the complexation of drugs in CD-NSs and the use of oxyresveratrol as an ingredient in nutraceutical products.
- PublicationOpen AccessSynergistic effect of cyclodextrins and methyl jasmonate on taxane production in Taxus x media cell cultures(Wiley, 2014-05-09) Sabater Jara, Ana Belén; Onrubia, Miriam; Moyano, Elisabeth; Bonfill, Mercedes; Palazón, Javier; Pedreño García, Mª Ángeles; Cusidó, Rosa M.; Biología VegetalMethyl jasmonate and cyclodextrins are proven effective inducers of secondary metabolism in plant cell cultures. Cyclodextrins, which are cyclic oligosaccharides, can form inclusion complexes with nonhydrophilic secondary products, thus increasing their excretion from the producer cells to the culture medium. In the present work, using a selected Taxus x media cell line cultured in a two-stage system, the relationship between taxane production and the transcript profiles of several genes involved in taxol metabolism was studied to gain more insight into the mechanism by which these two elicitors regulate the biosynthesis and excretion of taxol and related taxanes. Gene expression was not clearly enhanced by the presence of cyclodextrins in the culture medium and variably induced by methyl jasmonate, but when the culture was supplemented with both elicitors, a synergistic effect on transcript accumulation was observed. The BAPT and DBTNBT genes, which encode the last two transferases involved in the taxol pathway, appeared to control limiting biosynthetic steps. In the cell cultures treated with both elicitors, the produced taxanes were found mainly in the culture medium, which limited retroinhibition processes and taxane toxicity for the producer cells. The expression level of a putative ABC gene was found to have increased, suggesting it played a role in the taxane excretion. Taxol biosynthesis was clearly increased by the joint action of methyl jasmonate and cyclodextrins, reaching production levels 55 times higher than in nonelicited cultures.