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Cereus hildmannianus (Cactaceae) is a medicinal plant with various pharmacological and industrial applications. Plant biotechnology is an attractive approach for specialized metabolites production under controlled conditions, with the elicitation process being considered a highly effective strategy for increasing the production of bioactive compounds. In this regard, the present study investigated the effects of elicitors, sucrose (S, 1–8 g L− 1), UV-C light (UV-C, 10–80 min), salicylic acid (SA, 50–200 µmol), and jasmonic acid (JA, 50–200 µmol), on the profile of esterase isozymes, total phenolic and total flavonoid contents, antioxidant activity (FRAP, DPPH, and ABTS), and the dereplication method by Ultra-High-Performance Liquid Chromatography coupled with Mass Spectrometry (UHPLC-ESI-Q-TOF-MS/MS). The isozyme profile was not significantly changed. The greatest accumulations of total phenolics were observed in callus tissues induced with 100 µmol JA (390.8), 200 µmol SA (355.5), 20 min UV-C (182.9), and 6 g L− 1 S (122.1 µg mg− 1 DW); the greatest concentrations of total flavonoids were observed following culture with 200 µmol SA (89.8) and 2 g L− 1 S (28.4 µg mg− 1 DW). Furthermore, the dereplication study allowed putative identification of thirty-six compounds (thirty phenolic and organic acids, and six flavonoids). The greatest antioxidant activities were obtained with callus tissues induced with 100 µmol JA, 200 µmol SA, 80 min UV-C, and 1 g L− 1 S. The elicitors were able to alter the production of phenolic compounds in callus cultures of C. hildmannianus. These results are promising for the clean and sustainable bioproduction of bioactive molecules for pharmaceutical and cosmeceutical purposes, prioritizing the conservation of the species.

This study was to evaluate the biological activity of the extract of Botryosphaeria fabicerciana isolated from leaves of Morus nigra. The volatile compounds from the crude extract were analysed by GC-MS which demonstrate that mellein and β-orcinaldehyde were are the major compounds. The best minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the extract was observed against Gram-positive bacteria, with a MIC of 15.6 μg/mL towards B. cereus and MIC of 62.5 μg/mL towards S. aureus and B. subtilis. MBC values of 31.25 μg/mL, 62.5 μg/mL, and 250 μg/mL were observed towards B. cereus, B. subtilis, and S. aureus, respectively. The cytotoxicity analyses showed CC50 of 115 μg/mL. The crude extract showed antioxidant activity by the DPPH, ABTS, and FRAP assays. Therefore, the extract of the endophytic fungus presented biotechnological potential as an antibacterial and antioxidant agent.

β-ecdysone is the main compound of commercial interest produced by the roots of Pfaffia glomerata (Spreng.) Pedersen, and is used in herbal medicines and dietary supplements. In-vitro root culture facilitates the continuous production of high-quality compounds, and uses sustainable production techniques. Adventitious roots of P. glomerata were cultivated in a roller bottle system and an orbital gyratory system, both in the absence of light. The quantification of β-ecdysone by Liquid Chromatography-Mass Spectrometry/Mass Spectrometry (LC–MS/MS) revealed that the highest accumulation of metabolites occurred after 8 weeks in the roller bottle culture system, wherein 1.81 μg of β-ecdysone was recovered from 30 mg of dry root. These results suggest that adventitious root culture in-vitro may be an alternative strategy for β-ecdysone production, and demonstrates the potential use of bioreactors for practical applications.

Latex from Tabernaemontana catharinensis contains synthesized bioactive compounds known as monoterpenoid indole alkaloids (MIAs). Such compounds have several biological activities, including as cholinesterase inhibitors, and have analgesic, anti-inflammatory, bactericidal, antileishmanial, antitumoral andestrogenic properties, as well as acting as stimulants and depressants of the central nervous system. The objective of the present study was to perform the metabolite profiling of alkaloidal extracts from T. catharinensis latex. The alkaloidal fraction obtained from T. catharinensis latex was analyzed by Gas Chromatography coupled to Mass Spectrometry (GC–MS). The T. catharinensis latex was obtained after cutting the stem and allowing it to flow into tap water that was submitted to an acid-basic procedure for total MIA extraction, which was then analyzed by GC–MS. The total MIA extract was directly subjected to GC–MS without a derivatization step, allowing the detection and identification of ten known and three unknown MIAs. The described method resulted in effective separation and analysis by GC-EI-MS, which permitted the identification of known bioactive MIAs, as well as the structural elucidation of new MIAs with potential biological activities.

The endophytic fungi live inside plant tissues and do not develop any disease to their hosts. These microorganisms can produce bioactive compounds. In this work, we aimed to evaluate the leaf endophytic fungi microbiota from J. brandegeana using the culture-dependent method and multilocus sequence analysis. The endophyte's isolation was performed by the fragmentation method and the MLSA phylogeny was made using ITS, elongation factor 1-α, β-tubulin, glyceraldehyde-3-phosphate and, actin genes. For the screening selection, agar piece test was performed and, amongst the strains, JB06 isolate was chosen for metabolic extraction from the fermented culture and the evaluation of the antibacterial, antioxidant, antiproliferative activities and, phenolic compounds assay. Through the MLSA analysis, there were five genera of fungi identified, with Colletotrichum the higher isolated. For the antioxidant assay using the fungal extract obtained from Mycoleptodiscus indicus (JB06), the best results were obtained with the ABTS method. Antiproliferative effects by the crude extract of the endophyte against the tumor hepatocellular carcinoma cells (HepG2/C3A) were found. In summary, our findings revealed highly diverse endophytic fungi associated with J. brandegeana, with the bioprospection of the endophyte M. indicus with biotechnological potential for the health field.

Cereus hildmannianus is a cactus exhibiting morphological and physiological adaptation of its cladodes which ensuring growth in climatic and soil conditions unfavourable for many plant species. Currently, limited water resources and increasing demand for renewable energy make cacti a biomass source for the production of biofuels. Somaclones regenerated from callus in vitro can be a source of new raw material in useful plants. The objective of this work was to determine if the regenerated plants showing two different morphologies present polysaccharide composition different from the wild plant. Somaclones aqueous extraction shows the absence of soluble polysaccharides as mucilage. The alkaline extraction of in vivo cultivated plant showed the presence of starch, type I arabinogalactan, and arabinoxylan and the somaclones showed type I arabinogalactan and arabinoxylan in both morphologies. Hemicelluloses found in the somaclones are not different from in vivo cultivated plants, but somaclones not almost biosynthesize mucilage and starch.

The development of more efficient and environmentally friendly analytical methods represents a current focus for the fine chemical industry. In particular, microscale methodologies that are free of solvents/reagents. The headspace-GC/MS (HS-GC) methodology was employed in this study as a tool for monitoring a biocatalysed reaction of (4S)-(+)-carvone using Phoma sp., a filamentous fungus from human skin. Biocatalysis provides some advantages, such as high efficiency, high degrees of regioselectivity, chemoselectivity, and enantioselectivity. In order to optimize the small scale biocatalytic reaction of the (4S)-(+)-carvone by the filamentous fungus Phoma sp. was used headspace GC/MS methodology, factorial design of experiments and the response surface methodology (RSM) was performed using the biomass of the fungus, substrate mass and pH as parameters. It was observed that for all reactions conditions tested, forming the products (1 R,4S)-dihydrocarvone and (1S,4S)-dihydrocarvone. The most influential factor was pH, with the highest conversion rate (>95%) and diastereomeric excess (d.e.) (>80%) obtained at pH 5.0. Thus, it was demonstrated that human skin Phoma sp. fungus showed significant bioreduction activity and that headspace GC/MS is an efficient approach for real-time monitoring the biocatalysed reactions.

In this study, the effect of sucrose on the neutral lipid profile of adventitious root cultures of Stevia rebaudiana was evaluated. The cultures were obtained employing a roller bottle system. In this system, Schott-type flasks were used, which contained Murashige and Skoog liquid medium at 33.3% strength (MS/3) supplemented with 30, 60, and 80 g L-1 of sucrose, respectively, and 10.7 mM 1-naphthaleneacetic acid (NAA). The spectroscopic analyzes showed that the portion of polyunsaturated fatty acids (PUFAs) was highest in roots treated with 30 g L-1 of sucrose. The spectrometric analyzes showed that the palmitic acid was found to be present in relatively higher amounts in the roots submitted to the MS/3-30 g L-1 (31.9%) and MS/3-60 g L-1 (29.5%) sucrose treatments, and lower in the treatment with MS/3-80 g L-1 (28.8%) of sucrose. Also, the treatment using 30 g L-1 of sucrose was the best for obtaining unsaturated fatty acids (UFAs) in the culture, with a relative percentage of 62.9%. Our results indicate that the MS medium that received 30 g L-1 of sucrose induced a lesser abiotic stress condition, which favored PUFAs production in the adventitious root cultures of S. rebaudiana. En este estudio, se evaluó el efecto de la sacarosa sobre el perfil de lípidos neutros de cultivos de raíces adventicias de Stevia rebaudiana. Los cultivos se obtuvieron utilizando un sistema de botella giratoria. En este sistema se utilizaron matraces tipo Schott (con tapón), que contenían medio líquido Murashige y Skoog al 33,3% de fuerza (MS/3) suplementado con 30, 60 y 80 g L-1 de sacarosa, respectivamente, y 10,7 mM ácido de 1-naftalenoacético (ANA). Los análisis espectroscópicos de los extractos de raíces mostraron que la porción de ácidos grasos poliinsaturados (AGP) fue mayor en las raíces tratadas con 30 g L-1 de sacarosa. Los análisis espectrométricos mostraron que el ácido palmítico estaba presente en cantidades relativamente mayores en las raíces sometidas a los tratamientos con sacarosa MS/3-30 g L-1 (31.9%) y MS/3-60 g L-1 (29.5%), y menor en el tratamiento con MS/3-80 g L-1 (28,8%) de sacarosa. En el tratamiento con MS/3-80 g L-1 de sacarosa, el ácido esteárico fue el ácido graso predominante (56,8%). Además, el tratamiento con 30 g L-1 de sacarosa fue el mejor para la obtención de ácidos grasos insaturados (HUFA) en el cultivo, con un porcentaje relativo de 62,9%. Nuestros resultados indicaron que el medio MS que recibió 30 g L-1 de sacarosa indujo una condición de menor estrés abiótico, lo que favoreció la producción de HUFAs en los cultivos de raíces adventicias de S. rebaudiana. Neste estudo, o efeito da sacarose no perfil lipídico neutro de culturas de raízes adventícias de Stevia rebaudiana foi avaliado. As culturas foram obtidas usando um sistema de garrafa giratória. Neste sistema, foram utilizados frascos do tipo Schott que continham meio líquido de Murashige e Skoog de 33,3% de força (MS/3), suplementado com 30, 60 e 80 g L-1 de sacarose, respectivamente, e 10.7 mM de ácido 1-naftalenoacético (ANA). As análises espectroscópicas dos extratos das raízes demonstraram que a porção de ácidos graxos poli-insaturados (AGPIs) foi mais alta nas raízes tratadas com 30 g L-1 de sacarose. As análises espectrométricas mostraram que o ácido palmítico estava presente em quantidades relativamente maiores nas raízes submetidas com tratamento de sacarose de MS/3-30 g L-1 (31.9%) e MS/3-60 g L-1 (29.5%), e em quantidades menores no tratamento de sacarose de MS/3-80 g L-1 (28.8%). No tratamento de sacarose de MS/3-80 g L-1, o ácidos esteárico foi o ácido graxo prevalente (56.8%). Além disso, o tratamento usando 30 g L-1 de sacarose foi o melhor para obtenção de ácidos graxos insaturados nas culturas com porcentagem relativa de 62,9%. Nossos resultados indicaram que o meio MS que recebeu 30 g L-1 de sacarose induziu uma condição de menor estresse abiótico o que favoreceu a produção de AGPIs nas culturas de raízes adventícias de S. rebaudiana.

This study was to evaluate the biological activity of the extract of

In this work, the antibacterial activity of a crude extract of the endophytic fungus Flavodon flavus (JB257), isolated from leaves of Justicia brandegeana, was evaluated against both the vegetative and sporulated forms of Alicyclobacillus acidoterrestris. The microdilution technique was performed in order to determine the antibacterial activity of the crude extract alone as well as in combination with the bacteriocin, nisin. The minimum inhibitory concentration (MIC) of the crude extract and nisin alone against A. acidoterrestris vegetative forms were 250 μg/mL and 31.5 μg/mL, respectively, while the minimum bactericidal concentrations (MBC) were 1000 μg/mL and 62.5 μg/mL,respectively. For A. acidoterrestris spores, treatment with the crude extract at a concentration of 500 μg/mL caused a 47% reduction in growth, while nisin at 62.5 μg/mL could reduce 100% of the growth. The in vitro evaluation of the crude extract combined with nisin against A. acidoterrestris by the Checkerboard method showed a synergistic interaction between the two compounds. In addition, greater selectivity towards bacterial cells over host cells, a human hepatocyte cell line, was achieved when the crude extract was combined with nisin, Using scanning electron microscopy, interferences in the cell membrane of A. acidoterrestris could be observed after treatment with the crude extract. The results presented in this study indicate that the crude extract of the endophyte F. flavus has biotechnological potential in the food industry, especially for the treatment of orange juices through the control of A. acidoterrestris.

Ethnopharmacological relevance Cereus hildmannianus (K.) Schum. (syn. C. peruvianus) is a native medicinal plant in the Neotropical region. C. hildmannianus cladodes extracts are used in folk medicine for weight loss, reducing cholesterol, low-density lipoprotein (LDL) levels, as diuretic and cardiotonic, and to treat various diseases, including pulmonary disorders, rheumatism, and in topical treatment for wounds and lithiasis. Fruits and flowers of C. hildmmanianus have high nutritional value. Aim of the review: In this review, previous reports on C. hildmannianus (syn. C. peruvianus) concerning its botanical description, geographical distribution, ethnomedicinal use, phytochemistry, in vitro and in vivo pharmacological properties, food benefits and plant biotechnology were summarized. Materials and methods Scientific search engines, including ScienceDirect, Capes Journals Portal, Google Scholar, PubMed, Scielo, and Scifinder, were consulted to gather data on C. hildmannianus. The present review is an up-to-date and comprehensive analysis of phytochemical compounds, ethnomedicinal uses, phytopharmacological activities, and food value of cladodes, flowers, fruits, seeds, and tissue culture from C. hildmannianus. Results In traditional medicine, C. hildmannianus is used to treat various diseases, including pulmonary disorders, rheumatism, and in topical treatment for wounds and lithiasis. The extracts from the cladodes of C. hildmannianus exhibited numerous in vitro and in vivo pharmacological activities such as gastroprotective, antioxidant, antifungal, ovicidal, hemagglutinating and slimming, and anticancer activity. These extracts’ substances belong to different classes of chemical compounds such as fatty acids, polysaccharides, terpenes, alkaloids, phenolic acids, and flavonoids Conclusions Ethnomedicinal surveys have indicated the use of C. hildmannianus, an important medicinal plant in South America, for the treatment of various diseases. The ethnobotanical, phytochemical, pharmacological, and ethnomedicinal properties from various morphological parts of the plant of C. hildmannianus were highlighted in this review, which provides information for future studies, commercial exploration and reveals that this plant has a huge potential for pharmaceutical and nutraceutical applications.

Generally, the selection of fructans prebiotics and probiotics for the formulation of a symbiotic has been based on arbitrary considerations and in vitro tests that fail to take into account competitiveness and other interactions with autochthonous members of the intestinal microbiota. However, such analyzes may be a valuable step in the development of the symbiotic. The present study, therefore, aims to investigate the effect of lactobacilli strains and fructans (prebiotic compounds) on the growth of the intestinal competitor Klebsiella oxytoca, and to assess the correlation with short-chain fatty acids production. The short-chain fatty acids formed in the fermentation of the probiotic/prebiotic combination were investigated using NMR spectroscopy, and the inhibitory activities were assessed by agar diffusion and co-culture methods. The results showed that Lactobacillus strains can inhibit K. oxytoca, and that this antagonism is influenced by the fructans source and probably associated with organic acid production.

The synthesis of optically pure compounds is increasingly in demand among the pharmaceutical, fine chemical and agro-food industries, while the importance of chirality in the activity and biological properties of many compounds has previously been established. The aim of the present study was therefore to evaluate the biotransformation capacities of (+)-carvone and (−)-carvone using the fungi Scolecobasidium sp, three lines of Cladosporium sp, Phoma sp, Aureobasidium sp and Epicoccum sp, all obtained from human skin. The seven fungi evaluated were capable of hydrogenating the activated alkene, followed by the reduction of ketone to chiral alcohol, with conversions between 9.5 and 100%, and with diastereomer excess (d.e.) of over 89% of dihydrocarveol when (+)-carvone was used as a substrate. These results demonstrate that the filamentous fungi of human skin are potential biocatalytic tools for obtaining chiral alcohols.

Global interest in sugarcane has increased significantly in recent years because of its economic impact on sustainable energy production. The purpose of the present study was to evaluate changes in the concentrations of total sugars, amino acids, free proline, and total proteins by colorimetric analyses and nuclear magnetic resonance (NMR) to perform a metabolic profiling of a water-soluble fraction of symplastic sap in response to the constitutive expression of a mutant Δ1-pyrroline-5-carboxylate synthetase (P5CS) gene from Vigna aconitifolia. However, there was not a significant increase in the free proline content in the sap of transgenic plants compared to the non-transformed control plants. The most noticeable difference between the two genotypes was an almost two-fold increase in the accumulation of sucrose in the stem internodes of P5CS transgenic sugarcane plants. The results presented in this work showed that transgenic sugarcane plants with increased levels of free proline accumulates high soluble sugar content and, therefore, may represent a novel genotype for improving sugarcane cultivars.

The antiviral potential of natural polysaccharide compounds has been demonstrated, especially against enveloped viruses and members of the Herpesviridae family. Two polysaccharide fractions obtained from Stevia rebaudiana (Bertoni) leaves, that were active against Herpes simplex virus type 1 (HSV-1) were studied to investigate their mode of action. Both polysaccharides - SFW (crude faction) and SSFK (homogeneous alkaline fraction) - exerted antiviral effects on the initial stages of HSV-1 infection by inhibiting viral adsorption and penetration. When added after virus internalization, both fractions decreased plaque size. The effect of the fractions was confirmed by investigating viral glycoprotein expression. Based on the mode of action of the polysaccharides demonstrated in the present work and on their selectivity index, the polysaccharides obtained from S. rebaudiana could be an alternative treatment of infections caused by HSV-1.

Alicyclobacillus spp. is composed of Gram-positive, aerobic, thermoacidophilic, endospore-forming bacteria that cause food and beverage spoilage. The presence of Alicyclobacillus spp. may result in the production of guaiacol, which leads to sensory changes in the odour and taste of citrus juices and acidic foods. Tabernaemontana catharinensis (A. DC) is a plant belonging to the family Apocynaceae that produces milky latex with several biological activities described as antioxidant, antiviral, antimicrobial, trypanocidal and anti-leishmanicidal. Therefore, this study aims to evaluate the antimicrobial activity of the crude latex of T. catharinensis (A. DC) against microorganisms of the genus Alicyclobacillus spp. The minimum inhibitory concentration of latex was 7.81 μg/ml for the five Alicyclobacillus species analysed. The minimum bactericidal concentration for the species Alicyclobacillus acidoterrestris 0244T, A. hesperidum 0298T, A. acidiphilus 0247T and A. cycloheptanicus 0297T was 250 μg/ml. Cytotoxicity analysis demonstrated that latex was toxic to Vero cells at concentrations greater than 84.67 μg/ml. Scanning electron microscopy revealed changes in the cell wall of A. acidoterrestris 0244T present in orange juice when treated with crude latex. The results obtained suggest that the crude latex of T. catharinensis (A. DC) displays an antimicrobial effect against Alicyclobacillus, with potential for application in the food industry. El género Alicyclobacillus spp. és compuesto por bactérias Gram positivas, aerobias, en forma de bacilos termorresistentes, que tienen capacidad de formar esporas que funcionan como deterioro de alimentos y bebidas. El desarollo de estas bacterias puede resultar en la producción del compuesto Guaiacol, que tiene como consecuencia modificaciones sensoriales. La planta Tabernaemontana catharinensis (A.DC), perteneciente a la familia Apocynaceae, produce una sustancia llamada látex con actividad biológica descrita. El estudio del látex crudo de T. catharinensis (A. DC) frente a microorganismos del género Alicyclobacillus, tiene como objetivo investigar su actividad antimicrobiana. Las metodologías aplicadas fueron: determinación de concentración mínima inhibitoria (MIC) y concentración mínima bactericida (MBC), citotoxicidad y microscopía electrónica de barrido. Los resultados encontrados fueron: una MIC de 7.81 µg/mL para todas las bacterias analizadas; y un resultado de MBC más efectivo que ocurrió para las cepas A. acidoterrestris 0244T, A. hesperidum 0298T, A. acidiphilus 0247T y A. cycloheptanicus 0297T que presentaron MBC de 250 µg/mL. En cuanto a la cepa A. acidocaldarius 0299T, el MBC fue de 500 µg/mL. El análisis de citotoxicidad demostró que era tóxico para las células VERO en concentraciones superiores a 84,67 ± 8,14 µg/mL. En el microscopio electrónico de barrido fue posible observar alteraciones en la integridad de la pared de Alicyclobacillus acidoterrestris 0244T en jugo de naranja, cuando expuesto a látex crudo en diferentes concentraciones. Podemos concluir que el látex crudo de Tabernaemontana catharinensis (A. DC) p efecto antimicrobiano frente a Alicyclobacillus acidoterrestris 0244T. O gênero Alicyclobacillus spp. é composto por bactérias Gram-positivas, aeróbicas, na forma de bacilos, ácido termoresistentes, com capacidade de formar esporos e atuarem como deteriorante de bebidas e alimentos. O desenvolvimento dessas bactérias pode resultar na produção do composto Guaiacol que leva a alterações sensoriais. A planta Tabernaemontana catharinensis (A.DC) pertence à família Apocynaceae é produtora de uma substância chamada látex com atividade biológica descrita. O estudo do látex bruto de T. catharinensis (A. DC) frente aos microrganismos do gênero Alicyclobacillus tem como objetivo investigar sua atividade antimicrobiana. As metodologias desenvolvidas foram: determinação da concentração inibitória mínima (MIC) e concentração bactericida mínima (MBC), citotoxicidade e microscopia eletrônica de varredura. Os resultados encontrados foram MIC de 7,81 µg/mL para todas as bactérias analisadas e o resultado mais eficaz de MBC ocorreu para as cepas A. acidoterrestris 0244T, A. hesperidum 0298T, A. acidiphilus 0247T e A. cycloheptanicus 0297T que apresentaram MBC de 250 µg/mL. Já para a cepa A. acidocaldarius 0299T o MBC foi de 500 µg/mL. A análise de citotoxicidade demonstrou que foi tóxico em células Vero em concentrações superiores a 84,67 ± 8,14 µg/mL. No microscópio eletrônico de varredura foi possível observar alterações na integridade da parede de Alicyclobacillus acidoterrestris 0244T em suco de laranja, quando tratado com látex bruto em diferentes concentrações. Podemos concluir que o látex bruto de Tabernaemontana catharinensis (A. DC) apresenta efeito antimicrobiano frente a Alicyclobacillus acidoterrestris 0244T.

The Conyza sp. is an annual or short-lived perennial weed of the Asteraceae family. Fructans are linear fructose polymers used in the food industry as a prebiotic nutrient, fat, and sugar substitute and in the pharmaceutical industry as a drug delivery system. The objective of this study was to chemically characterize preliminarily fructans from Conyza sp. roots: fructooligosaccharides (FOS) and inulin-type and evaluated the prebiotic potential activity of these molecules in vitro. Chemical analyses (1H NMR, 13C NMR, GC-MS, FT-IR, and off-line ESI-HRMS) confirmed the presence of the fructans molecules in the aqueous crude extracts of Conyza sp. roots. The study of the prebiotic effect of Conyza sp. fructans in vitro showed an ability to stimulated lactobacilli strains with growth slightly similar (Conyza FOS) and smaller (Conyza inulin) than the commercial prebiotic inulin available (Orafti® GR). These data suggest that Conyza sp. a weed that causes problems agronomic can be used as a promising source to obtain fructans, compounds with potential in the food and technological industries.

Stevia rebaudiana (Bertoni) Bertoni, Asteraceae, is used as a food additive because its leaves are a source of steviol glycosides. There are examples of tissue culture based on micropropagation and phytochemical production of S. rebaudiana leaves but there are few studies on adventitious root culture of S. rebaudiana. More than 90% of the plants used in industry are harvested indiscriminately. In order to overcome this situation, the development of methodologies that employ biotechnology, such as root culture, provides suitable alternatives for the sustainable use of plants. The aim of this study was to compare morpho-anatomical transverse sections of S. rebaudiana roots grown in vitro and in vivo. The in vitro system used to maintain root cultures consisted of a gyratory shaker under dark and light conditions and a roller bottle system. Transverse sections of S. rebaudiana roots grown in vitro were structurally and morphologically different when compared to the control plant; roots artificially maintained in culture media can have their development affected by the degree of media aeration, sugar concentration, and light. GC–MS and TLC confirmed that S. rebaudiana roots grown in vitro have the ability to produce metabolites, which can be similar to those produced by wild plants. Keywords: Stevia rebaudiana, Adventitious root cultures, Morpho-anatomical, Chemical profile

Stevia rebaudiana (Bertoni) is widely studied because of its foliar steviol glycosides. Fructan-type polysaccharides were recently isolated from its roots. Fructans are reserve carbohydrates that have important positive health effects and technological applications in the food industry. The objective of the present study was to isolate and characterize fructo-oligosaccharides (FOSs) from S. rebaudiana roots and in vitro adventitious root cultures and evaluate the potential prebiotic effect of these molecules. The in vitro adventitious root cultures were obtained using a roller bottle system. Chemical analyses (gas chromatography-mass spectrometry, (1)H nuclear magnetic resonance, and off-line electrospray ionization-mass spectrometry) revealed similar chemical properties of FOSs that were obtained from the different sources. The potential prebiotic effects of FOSs that were isolated from S. rebaudiana roots enhanced the growth of both bifidobacteria and lactobacilli, with strains specificity in their fermentation ability.

Inulin and fructooligosaccharides (FOS) are fructans a reserve polysaccharides that have a great economic interest in the food and pharmaceutical industries. Cichorium endivia, also known as “escarole”, belongs to the family Asteraceae and normally is used as a salad, forage crops and raw material for the production of fructose. The objective of this work was to extract, characterize chemically, and evaluate the prebiotic activity in vitro from C. endivia roots FOS. By colorimetric assays, we to determined its chemical composition with 85.4% of total sugars containing 72.2% of fructose. HSQC-Q NMR was used to determine the monosaccharide composition. The NMR and off-line ESI-MS analysis showed that C. endivia FOS has a degree of polymerization equal to 6 and 7, respectively. In vitro prebiotic test showed that the same was fermented by all probiotic bacteria evaluated and the results showed that C. endivia FOS is a promising source of prebiotic fructans.

Biocatalysis represents an efficient and versatile tool for organic synthesis and green chemistry, since enzymes can perform many chemical transformations with advantages over their chemical counterparts. (4R)-(−)-carvone and its reduced derivatives are natural products with biological and industrial applications, specially (1R,2S,4R)-neodihydrocarveol, due to its pharmacological properties, as well as fragrances supplies. Dialkyl imidazolium-based ionic liquids (ILs) are known to be useful in biocatalytic processes due to their unique physico-chemical properties, low toxicity and ability to improve enzymatic activity. In the present study, we describe the bioreduction of (4R)-(−)-carvone using a filamentous fungus isolated from human skin, molecularly identified by MLSA as Epicoccum sorghinum, and ionic liquid (IL) 1-butyl-3-methyl-imidazolium tetrafluoroborate ([BMIM][BF4]), as co-solvent. The addition of 0.2% (v/v) of [BMIM][BF4] favoured the bioreduction of the substrate (4R)-(−)-carvone at a concentration of 0.198 mmol, affording 65% of the major compound (1R,2S,4R)-neodihydrocarveol, with diastereomeric excess greater than 99% after 18 h of biocatalytic reaction. Also, the isolated product was obtained with an appreciable yield of 59% after column chromatography.

The antiviral potential of natural polysaccharide compounds has been demonstrated, especially against enveloped viruses and members of the

Several species of Aspidosperma plants are used to treat diseases in the tropics, including Aspidosperma ramiflorum, which acts against leishmaniasis, an activity that is experimentally confirmed. The species, known as guatambu-yellow, yellow peroba, coffee-peroba and matiambu, grows in the Atlantic Forest of Brazil in the South to the Southeast regions. Through a guided biofractionation of A. ramiflorum extracts, the plant activity against Plasmodium falciparum was evaluated in vitro for toxicity towards human hepatoma G2 cells, normal monkey kidney cells and nonimmortalised human monocytes isolated from peripheral blood. Six of the seven extracts tested were active at low doses (half-maximal drug inhibitory concentration < 3.8 µg/mL); the aqueous extract was inactive. Overall, the plant extracts and the purified compounds displayed low toxicity in vitro. A nonsoluble extract fraction and one purified alkaloid isositsirikine (compound 5) displayed high selectivity indexes (SI) (= 56 and 113, respectively), whereas compounds 2 and 3 were toxic (SI < 10). The structure, activity and low toxicity of isositsirikine in vitro are described here for the first time in A. ramiflorum, but only the neutral and precipitate plant fractions were tested for activity, which caused up to 53% parasitaemia inhibition of Plasmodium berghei in mice with blood-induced malaria. This plant species is likely to be useful in the further development of an antimalarial drug, but its pharmacological evaluation is still required.

Pfaffia glomerata (Amaranthaceae) is popularly known as “Brazilian ginseng.” Previous studies have shown that fructose is the major carbohydrate component present in its roots. Inulin-type fructans, polymers of fructose, are the most widespread and researched prebiotics. Here, we isolated and chemically characterized inulin extracted from P. glomerata roots and investigated its potential prebiotic effect. Fructans were isolated and their structures were determined using colorimetric, chromatography, polarimetry, and spectroscopic analysis. The degree of polymerization (DP) was determined, and an in vitro prebiotic test was performed. The structure of inulin was confirmed by chromatography and spectroscopic analysis and through comparison with existing data. Representatives from the genera Lactobacillus and Bifidobacterium utilized inulin from P. glomerata, because growth was significantly stimulated, while this ability is strain specific. The results indicated that inulin extracted from P. glomerata roots represents a promising new source of inulin-type prebiotics.

Cereus peruvianus Mill. is a cactus species that belongs to the family Cactaceae, popularly known as “Mandacaru”. It presents several pharmacological activities in popular medicine. Its extract is used to reduce total cholesterol and low-density lipoprotein (LDL) levels and consequently reduce the risk of cardiovascular disease. It also has antineoplastic actions related to the presence of polyunsaturated fatty acids. The in vitro tissue culture of C. peruvianus occurs under controlled environmental conditions, is independent of climatic or soil changes, and is free from microorganism and insect contamination. These advantages make it a sustainable biotechnology alternative to achieve specific metabolites, such as fatty acids. Tissue culture lipids were extracted from maceration and separated into neutral lipids, glycolipids, and phospholipids by column chromatography. The analysis of these fractions by gas chromatography and 1H and 13C nuclear magnetic resonance spectroscopy showed that the cell cultures had high amounts of unsaturated fatty acids, such as ω-3, ω-6, eicosapentaenoic acid, and docosahexaenoic acid. Biologic assays were used to investigate potential antitumor activity using the sulforhodamine B method, and we observed a 50% cytotoxic concentration of 250 μg/ml. The results contribute to knowledge about the lipid profile and antitumor activity of callus cultures of C. peruvianus.

The polysaccharide inulin has great importance in the food and pharmaceutical industries. The degree of polymerization (DP) of inulin influences important properties, such as, solubility, thermal stability, sweetness power and prebiotic activity. Molecules with a high degree of polymerization are obtained through physical techniques for enrichment of the inulin chains because they are not commonly obtained from plants extract. Gas chromatography/Mass Spectrometry and 1 H Nuclear Magnetic Resonance analysis showed that inulin from Stevia rebaudiana roots has a degree of polymerization (DP n 28) higher than the value of DP n 12–15 for inulins from other plant species. Furthermore, the methodology of freeze/thaw to enrich the chains allowed us to increase the DP, similarly to other methodologies used for the enrichment of inulin chains. The prebiotic assays confirm that inulin from S. rebaudiana has a high DP. The combined use of these molecules with low degree of polymerization fructans seems to be advantageous to prolong the prebiotic effect in the colon. Our results suggest that S. rebaudiana roots are a promising source of high degree polymerization inulins.

In the current study we reported cultivation, extraction procedure, analysis and preliminary characterization of the aqueous extract from Cereus peruvianus callus culture and evaluated its anti ulcerogenic activity in vivo models of experimental ulcers in Wistar rats. The obtained aqueous extract from callus (AC) was dialyzed and subjected to freeze-thaw process, providing a possible polysaccharide. The carbohydrate and protein contents of the aqueous extract were estimated at 53.4% and 0.66%, respectively, composed primarily of galactose, arabinose and galacturonic acid, with minor amounts of glucose. This appeared heterogeneous when analyzed by high-performance size-exclusion chromatography and a multiangle laser light scattering detector (HPSEC-MALLS). The AC was found to be significantly effective against ethanolinduced lesions but was ineffective against indomethacin-induced lesions. The callus culture of C. peruvianus is an alternative source for the synthesis of substances originally produced by plants. The calluses grown indefinitely in vitro under controlled conditions are stable tissues, and the aqueous extract from calluses may be used instead of fully developed plants using the protocols described in this study.

Cell wall polysaccharides from leaves of Stevia rebaudiana were extracted successively with water and with aq. 10% KOH. After the purification steps, homogeneous fractions (SFW-10RM and SSFK-10RM) were analyzed by sugar composition, HPSEC, methylation and 13C NMR spectroscopy analysis. The results showed that SFW-10RM is a pectic arabinogalactan with an unusual β-(1→6)-linked D-Galp residues forming the main chain. Approximately 38% of the β-d-Galp units of the backbone carry branches on position O-3, consisting of single D-Galp units or arabinan side chains. Arabinose residues were found to occupy mostly the terminal positions in both furanose and pyranose forms and as 2-, 5- and 3,5-linked residues in these side chains. Fraction SSFK-10RM is a similar arabinogalactan, differing mainly in the relative proportions of arabinans attached to the galactan core and in the content of D-GalpA residues present in the pectic domain. The crude aqueous and alkaline extracts and homogeneous SSFK-10RM showed antiviral activity against Herpes Simplex Virus type-1 (HSV-1) in vitro.

The present study was designated to evaluate semi-quantitative antileishmanial activity of alkaloidal extracts that were obtained from 1g of different parts of Aspidosperma ramiflorum (leaves, roots, seeds, and stem barks). Alkaloidal extracts of barks and leaves presented a good activity against the extracellular form (promastigotes) of Leishmania (L.) amazonensis. It is known that compounds responsible for the antileishmanial activity in the alkaloidal extracts from A. ramiflorum are the monoterpenoid indole alkaloids ramiflorine A and ramiflorine B, therefore extracts obtained from different plant parts were analyzed by electrospray ionization mass spectrometry (ESI-MS) in order to evidence the presence of these bioactive alkaloids. Based on these findings, alkaloidal extract from leaves was fractionated on preparative thin-layer chromatography in a bioassay-guided fractionation affording individual purified ramiflorines A and B. Both ramiflorines A and B showed significant activity against Leishmania (L.) amazonensis (LD(50) values of 18.5±6.5μg/ml and 12.63±5.52μg/ml, respectively). Our results are showing that alkaloidal extract from leaves is a promising alternative to the use of stem barks from A. ramiflorum.

Fructooligosaccharides have been isolated from roots and leaves of Stevia rebaudiana, by hot aqueous extraction, followed by precipitation with ethanol. Their structure has been determined using methylation and NMR analysis, MALDI-TOF, and ESI-MS. Fructooligosaccharides contained almost exclusively (2→1)-linked β-fructofuranosyl, with terminal α-glucopyranosyl and β-fructofuranosyl units. MALDI-TOF and ESI-MS analyses showed the wide range of degree of polymerisation (DP) present in various extracts. From roots and leaves, three different fractions gave profiles of homologous series, with DPs ranging up to 17 with MALDI-TOF and 19 using ESI-MS. These inulin-type fructooligosaccharides were the major component of extracts from S. rebaudiana roots and significant components from the leaves.

Cultures of adventitious roots of Stevia rebaudiana (Bert.) Bertoni were performed in a roller bottle system for the production of both primary and secondary metabolites. Adventitious roots were induced from 1-cm-long root tip explants derived from in vitro regenerated plantlets on solid Murashige and Skoog (MS 1962) media supplemented with 10.7 μM of α-naphthaleneacetic acid. These cultures were successfully maintained in the same medium for 6 months with regular subcultures after 4 weeks. Thereafter, the roots were cut into 1.0- to 1.5-cm-long segments and transferred to the roller bottle system containing a fresh root tissue culture on liquid MS medium supplemented with 10.7 μM NAA. The apparatus consisted of a flask rolling system adjusted to 4g, and 3° of flask inclination. The roots were allowed to grow in the absence of light for adaptation and adventitious root formation. The best conditions for cultivation were investigated, considering culture volume (25 ml), culture period (4 weeks), salt concentrations in the nutrient medium (33%) and optimal initial inoculum (0.2 g) of S. rebaudiana roots. These results could give important information on how to improve the development of adventitious roots of S. rebaudiana for the production of primary and secondary metabolites.