Secondary Metabolite Profiles Research Articles

Bread Enriched With Legume Microgreens and Leaves-Ontogenetic and Baking-Driven Changes in the Profile of Secondary Plant Metabolites.

Flavonoids, carotenoids, and chlorophylls were characterized in microgreens and leaves of pea (Pisum sativum) and lupin (Lupinus angustifolius) as these metabolites change during ontogeny. All metabolites were higher in the leaves for both species. Acylated quercetin and kaempferol sophorotrioses were predominant in pea. Genistein and malonylated chrysoeriol were predominant in lupin. Further, the impact of breadmaking on these metabolites using pea and lupin material of two ontogenetic stages as an added ingredient in wheat-based bread was assessed. In “pea microgreen bread” no decrease of quercetin was found with regard to the non-processed plant material. However kaempferol glycosides showed slight decreases induced by the breadmaking process in “pea microgreen bread” and “pea leaf bread.” In “lupin microgreen bread” no decrease of genistein compared to the non-processed plant material was found. Chrysoeriol glycosides showed slight decreases induced by the breadmaking process in “lupin microgreen bread” and “lupin leaf bread.” In all breads, carotenoids and chlorophylls were depleted however pheophytin formation was caused. Thus, pea and lupin microgreens and leaves are suitable, natural ingredients for enhancing health-promoting secondary plant metabolites in bread and may even be used to tailor bread for specific consumer health needs.

Secondary metabolite profiling, cytotoxicity, anti-inflammatory potential and in vitro inhibitory activities of Nardostachys jatamansi on key enzymes linked to hyperglycemia, hypertension and cognitive disorders

Ethnopharmacological relevanceNardostachys jatamansi (D. Don) DC., ‘Spikenard’ or ‘Jatamansi’, a highly valued, aromatic herb from alpine Himalayas has a long history of use as ethnomedicine and dietary supplements in Ayurveda, Unani and Chinese system of medicine since Vedic ages (1000–800 BC). In Ayurveda and traditional system of medicine, the species is used as stimulant, sedative, brain tonic or mind rejuvenator, antidiabetic, cardio tonic, and in the treatment of various neurological disorders such as insomnia, epilepsy, hysteria, anxiety and depression. It is considered as Sattvic herb in Ayurveda and is now commercially marketed either as single or poly-herbal formulations by many companies in national and international markets. Aim of the studyThe species has become threatened in its natural habitats due to over exploitation and illegal trade of its rhizomes for drug preparation in herbal and pharmaceutical industries. Considering the increasing demand and tremendous medicinal importance of this threatened plant species, a detailed study was undertaken to evaluate its antioxidant potential, secondary metabolite profiling, cytotoxicity, anti-inflammatory potential and in vitro enzyme inhibitory activities on key enzymes linked to hyperglycemia, hypertension and cognitive disorders in different plant parts of wild and in vitro-raised plants with respect to different solvent systems for its sustainable utilization. Materials and methodsAnti-cholinesterase activity of leaves and rhizome of wild and cultured plant extracts was investigated against both acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) enzymes. In vitro anti-hyperglycemic (α-amylase and PTP1B), anti-hypertensive (angiotensin-converting enzyme), anti-tyrosinase and anti-inflammatory potential (5-lipoxygenase and hyaluronidase) of different plant parts of wild and in vitro-raised plants with respect to different solvent systems were also evaluated. In vitro cytotoxic effect of rootstock extracts of wild and in vitro-derived plants were against cancer (HCT-116, MCF-7 and OE33) and two normal (HEK and MEF) cell lines. Secondary metabolite profiling of rhizome segments of wild and in vitro-derived plants was carried out by quantitative gas chromatography-mass spectrometry (GC–MS). ResultsIn vitro-raised plantlets showed comparative higher yield of various secondary metabolites with a significantly high antioxidant activity as compared to the wild plants. Methanolic rootstock extracts of both wild and in vitro-derived plants of N. jatamansi exhibited significant AChE (IC50 36.46 ± 2.1 and 31.18 ± 2.6 µg/ml, respectively) and BuChE (IC50 64.6 ± 3.5 and 60.12 ± 3.6 µg/ml, respectively) inhibitory potential as compared to standard inhibitor galanthamine (IC50 0.94 ± 0.03 and 4.45 ± 0.5 µg/ml). Methanolic rootstock extract of in vitro-derived plants showed significant α-amylase (IC50 90.69 ± 2.1 µg/ml), PTP1B (IC50 24.56 ± 0.8 µg/ml), angiotensin-converting enzyme (IC50 42.5 ± 3.6 µg/ml) and tyrosinase (IC50 168.12 ± 3.6 µg/ml) inhibitory potential as compared to standard acarbose (IC50 52.36 ± 3.1 µg/ml), ursolic acid (IC50 5.24 ± 0.8 µg/ml), captopril (IC50 32.36 ± 2.5 µg/ml) and kojic acid (IC50 = 54.44 ± 2.3 µg/ml). Both the methanolic rootstock and leaf extracts of tissue culture-derived plants exhibited promising anti-5-LOX and anti-hyaluronidase activities against the known inhibitor of 5-LOX and hyaluronidase. Furthermore, methanolic rootstock extracts of both wild and in vitro-derived plants exhibited promising cytotoxic effects to HCT-116, MCF-7 and OE33 cell lines as compared to the normal HEK and MEF after 12 h of treatment. Secondary metabolite profiling of wild and in vitro-derived plants by quantitative GC–MS analysis revealed the presence of different classes of terpenoids and phenolic acids might be responsible for its effective biological activities. ConclusionIn vitro-derived plants revealed a substantial anti-cholinesterases, anti-hyperglycemic anti-inflammatory, anti-hypertensive and anti-tyrosinase potential with higher yield of various bioactive metabolites and significantly higher antioxidant activity which substantially explain medicinal importance of N. jatamansi in traditional medicine, used for centuries in different Ayurvedic formulations. The present findings suggest that cultured plants could be a promising alternative for the production of bioactive metabolites with comparative biological activities to the wild plants.

Evaluation of antioxidant and neuroprotective activities of Cassia fistula (L.) using the Caenorhabditis elegans model.

BackgroundCassia fistula (L.) (Fabaceae) is a medicinal plant from tropical Asia. It is known for its marked antioxidant activity, which is attributed to its high phenolic content. The present study aims at testing both the antioxidant and neuroprotective effects of a hydroalcoholic extract from the aerial parts of Cassia fistula using the Caenorhabditis elegans model, which is widely used in this context.MethodsChemical profiling of secondary metabolites that seem to be responsible for both antioxidant and neuroprotective capacities was carried out by HPLC/PDA/ESI-MSn. Antioxidant activity was tested in vitro by CUPRAC and DPPH assays. In vivo antioxidant and neuroprotective activities were investigated using the C. elegans model.ResultsThe Cassia extract improved the survival rate of the nematodes and protected them against oxidative stress. In addition, a decrease in the accumulation of reactive oxygen species (ROS) was observed. The important role of DAF-16/FOXO pathway was confirmed through an increased nuclear localization of the DAF-16 transcription factor, increased expression of SOD-3 stress response gene and decreased expression of HSP-16.2. Furthermore, the putative involvement of SKN-1/NRF2 pathway was demonstrated by a decrease in GST-4 levels. A neuroprotective activity of the Cassia extract was shown by a decline in polyglutamine (polyQ40) aggregate formation and a delay in paralysis caused by amyloid beta (Aβ1–42) accumulation.DiscussionThe Cassia extract exhibits substantial antioxidant and neuroprotective activities in vivo, which might provide a rich and novel source of natural antioxidants and neuroprotective compounds to be further studied for the use in various food and cosmetic industrial fields.

Secondary metabolite production by cereal-associated penicillia during cultivation on cereal grains.

Cereals are vulnerable substrates for fungal growth and subsequent mycotoxin contamination. One of the major fungal genera to colonize the ecosystem of stored grain is Penicillium, especially species in the series of Viridicata and Verrucosa. Culturing these species on grains, we hoped to induce the production of relevant secondary metabolites produced by these fungi in the early stage of cereal breakdown. In a multivariate setup six different cereal grains (wheat, rye, barley, oat, rice, and maize), one kind of white beans, and two standard fungal media, Yeast Extract Sucrose agar (YES agar) and Czapek Yeast Autolysate agar (CYA agar), were inoculated with the ten most important cereal-associated species from Penicillium (P. aurantiogriseum, P. cyclopium, P. freii, P. melanoconidium, P. neoechinulatum, P. polonicum, P. tricolor, P. viridicatum, P. hordei, and P. verrucosum). P. nordicum is a meat-associated species, which was included due to its chemical association with P. verrucosum, in addition to see if a substrate change would alter the profile of known chemistry. We found that cereals function very well as substrates for secondary metabolite production, but did not present significantly different secondary metabolite profiles, concerning known chemistry, as compared to standard laboratory agar media. However, white beans altered the semi-quantitative secondary metabolite profiles for several species. Correlations between substrates and certain metabolites were observed, as illuminated by principal component analysis. Many bioactive secondary metabolites were observed for the first time in the analyzed fungal species, including ergot type alkaloids in P. hordei.

Response surface methodology to optimize supercritical carbon dioxide/co-solvent extraction of brown onion skin by-product as source of nutraceutical compounds

Food industry produces a large amount of onion wastes. Due to the high amount of bioactive compounds in onion by-products an idea for their reuse, could be use them as source of high-value functional and health ingredients. In this study, outer dry layers of coppery onion “Ramata di Montoro” were used as source of bioactive compounds. Firstly, the chemical profile of secondary metabolites of exhaustive extract, obtained by ultrasound assisted extraction was established by UHPLC-UV-HRMS/MS analysis. Subsequently, the supercritical fluid extraction was used as alternative and green method to recover flavonoids from onion skin. Main parameters such as pressure, temperature and composition of solvent modifier were optimized in order to improve the extraction efficiency of SFE technique, by using a response surface Box–Behnken design.

Abstract 279: Cranberry proanthocyanidins mitigate reflux-induced transporter dysregulation in esophageal adenocarcinoma

Abstract Esophageal adenocarcinoma (EAC) incidence rates have increased sharply (>500%) throughout the Westernized world since the 1980s, despite the widespread use of endoscopy and anti-reflux medications. Reasons for the rapid increase in EAC are still being unraveled; however, persistent, symptomatic reflux of gastric and duodenal contents, known as gastroesophageal disease (GERD), is considered the strongest risk factor. Our laboratory utilizes the rat esophagogastroduodenal anastomosis (EGDA) surgical model of reflux-induced EAC to evaluate mechanisms by which cranberry proanthocyanidins (C-PAC) delivered in the drinking water inhibit reflux-induced EAC. Findings show that C-PAC inhibits EAC formation by 83% with concomitant restoration of the gut microbial profile and significant reduction of primary and secondary bile acid metabolites in the esophagus of animals with reflux-inducing surgery, but through unknown mechanisms. Herein, we investigated whether transporters, important in bile acid transport, buffering capacity, excretion, and even resistance to anticancer agents, were restored by C-PAC in the context of reflux. Additionally, we utilized a publicly available GEO dataset (GSE37203) to assess transporter expression levels in patients with metaplastic Barrett's esophagus (BE) versus those with high grade dysplasia (HGD) or EAC for translational relevance. Methods included esophageal RNA isolation, transcriptome sequencing using the Illumina HiSeq platform, followed by validation utilizing the PrimePCR Drug Transporter array plate and database mining. Results show significant changes in solute carriers (SLC), ATP-binding cassette (ABC) and aquaporin transporters in the rat reflux-induced model for EAC with C-PAC mitigating alterations. EGDA altered one or more members in 71% of the SLC families with frequent changes observed in SLC25, SLC4, SLC35, SLC2 and SLC6. C-PAC favorably impacted >40% of the EGDA-induced SLC changes. A number of ABC transporters involved in glutathione transport and also implicated in drug resistance were altered by EGDA including Abcc1, Abcc3, Abcc4, Abcc5 and Abcg2 and C-PAC mitigated these changes. Aquaporins, Aqp3 and Aqp4, were significantly modulated in EGDA and restored with C-PAC. Significant differences in ABCC9, ABCC10, AQP6 and AQP9, in addition to several members of the SLC25, SLC4, SLC35, SLC2 and SLC6 families, were observed in patients with HGD/EAC compared to those with BE metaplasia. Therefore, altered expression of transporters following reflux-inducing surgery or GERD is likely a defensive mechanism by which cells attempt to adapt or protect from injurious bile acid exposure. Further research is warranted to investigate agents that restore normal transporter function, which may serve to concomitantly improve epithelial barrier function and mucosal integrity in the context of esophageal cancer progression. Citation Format: Katherine M. Weh, Bridget A. Tripp, Jennifer L. Clarke, Amy B. Howell, Jules Lin, David G. Beer, Andrew C. Chang, Laura A. Kresty. Cranberry proanthocyanidins mitigate reflux-induced transporter dysregulation in esophageal adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 279.

Comparison of volatile compounds released by entomopathogenic fungi

Entomopathogenic fungi are fungal species which are used as a potential source of biopesticides. These fungi produce secondary metabolites which in insects can cause disruption in the normal functioning of their bodies, disease or even death. In order to fully characterize the physiology of entomopathogenic fungi we should identify the volatile organic compounds which are involved in this process. Therefore, we conducted a qualitative and quantitative analysis of volatile compounds produced by entomopathogenic fungi. Seven different species of fungi were analyzed: Metarhizium anisopliae, Metarhizium flavoviride, Pandora sp., Isaria fumosorosea, Hirsutella danubiensis, Batkoa sp. and Beauveria bassiana. The analyses were performed using the HS-SPME/GCMS technique. In the analyzed fungi, 63 volatile compounds were identified and classified into the following groups: aldehydes, ketones, alcohols, esters, acids, terpenes and others. The results show that entomopathogenic fungi produce a wide profile of secondary metabolites. Principal Components Analysis was used to determine whether separate classes of fungi can be distinguished from one another based on their metabolite profiles.

Banksialactones and Banksiamarins: Isochromanones and Isocoumarins from an Australian Fungus, Aspergillus banksianus.

Chemical investigation of an Australian fungus, Aspergillus banksianus, led to the isolation of the major metabolite banksialactone A (1), eight new isochromanones, banksialactones B-I (2-9), two new isocoumarins, banksiamarins A and B (10 and 11), and the reported compounds, clearanol I (12), dothideomynone A (13), questin (14), and endocrocin (15). The structures of 1-11 were established by NMR spectroscopic data analysis, and the absolute configurations were determined from optical rotations and ECD spectra in conjunction with TD-DFT calculations. The secondary metabolite profile of A. banksianus is unusual, with the 11 most abundant metabolites belonging to a single isochromanone class. Conjugation of 1 with endocrocin, 5-methylorsellinic acid, 3,5-dimethylorsellinic acid, mercaptolactic acid, and an unknown methylthio source gave rise to five unprecedented biosynthetic hybrids, 5-9. The isolated compounds were tested for cytotoxicity, antibacterial, and antifungal activities, with hybrid metabolites 7-9 displaying weak cytotoxic and antibiotic activities.

A Bcl-2 Associated Athanogene (bagA) Modulates Sexual Development and Secondary Metabolism in the Filamentous Fungus Aspergillus nidulans.

The Bcl-2 associated athanogene (Bag) family is a multifunctional group of proteins distinguished by a conserved region known as the Bag domain (BD). Herein, we discuss the discovery and characterization of a Bag protein in the model genetic fungus Aspergillus nidulans, we designated BagA. BagA shares striking similarities in 3D structure, domain organization, amino acid properties, and Hsp70 binding surfaces to animal and plant Bags. While Hsp70 binding is a common feature of Bag proteins, our experimental evidence shows that BagA does not cooperate with A. nidulans Hsp70s, suggesting this association may not be a universal feature of Bag proteins. Gene expression of bagA was strongly induced during sexual development suggesting a role in developmental processes. Accordingly, the deletion of bagA (ΔbagA) negatively impacted sexual development, while its overexpression resulted in constitutive induction of sexual fruiting bodies and spores. Asexual and sexual development was linked to secondary metabolism in A. nidulans. Our data show that the deletion of bagA also provoked an altered secondary metabolite (SM) profile in both sexual and vegetative growth phases. Indeed, LC-MS analysis showed a significant enrichment of SMs in ΔbagA, including novel metabolites not produced by wild type strain. Enrichment of SMs in ΔbagA strain is particularly intriguing and suggest that altering cellular homeostasis can be used as a provocative strategy to activate cryptic metabolites and uncover novel bioactive compounds. Overall, our results indicate that Bag proteins in filamentous fungi share developmental regulatory roles with their animal and plant counterparts. We also show a potentially unique role for BagA in modulating secondary metabolism in A. nidulans. To our knowledge, this study provides a first insight into Bag function in filamentous fungi.

Comprehensive Secondary Metabolite Profiling Toward Delineating the Solid and Submerged-State Fermentation of Aspergillus oryzae KCCM 12698.

Aspergillus oryzae has been commonly used to make koji, meju, and soy sauce in traditional food fermentation industries. However, the metabolic behaviors of A. oryzae during fermentation in various culture environments are largely uncharacterized. Thus, we performed time resolved (0, 4, 8, 12, 16 day) secondary metabolite profiling for A. oryzae KCCM 12698 cultivated on malt extract agar and broth (MEA and MEB) under solid-state fermentation (SSF) and submerged fermentation (SmF) conditions using the ultrahigh performance liquid chromatography-linear trap quadrupole-ion trap-mass spectrometry (UHPLC-LTQ-IT-MS/MS) followed by multivariate analyses. We observed the relatively higher proportions of coumarins and oxylipins in SSF, whereas the terpenoids were abundant in SmF. Moreover, we investigated the antimicrobial efficacy of metabolites that were extracted from SSF and SmF. The SSF extracts showed higher antimicrobial activities as compared to SmF, with higher production rates of bioactive secondary metabolites viz., ketone-citreoisocoumarin, pentahydroxy-anthraquinone, hexylitaconic acid, oxylipins, and saturated fatty acids. The current study provides the underpinnings of a metabolomic framework regarding the growth and bioactive compound production for A. oryzae under the primarily employed industrial cultivation states. Furthermore, the study holds the potentials for rapid screening and MS-characterization of metabolites helpful in determining the consumer safety implications of fermented foods involving Koji mold.

Comparative metabolomic responses to gibberellic acid and 6-benzylaminopurine in Cunila menthoides Benth. (Lamiaceae): a contribution to understand the metabolic pathways.

Gibberellic acid elicited synthesis of many phenols from different classes and enhanced production of sesquiterpenoids, polyterpenoids, steroids and monoterpenoids compared to control and 6-benzylaminopurine. Little is known about the effects of 6-benzylaminopurine (BA) and gibberellic acid (GA3) on the synthesis of secondary metabolites in species of Lamiaceae. In this study, for the first time, the profile of secondary metabolites in plantlets of Cunila menthoides was characterized, using UPLC-ESI-Qq-oaTOF-MS. Ninety metabolites were identified, including polyphenols and terpenes. BA down-regulated most of the identified molecules in relation to GA3 and MS0 (control). The results showed that GA3 elicited synthesis of many phenols from different classes, and seemed to play a major role in the shikimate pathway in relation to BA. GA3 enhanced production of sesquiterpenoids, polyterpenoids, steroids and monoterpenoids compared to MS0 and BA, and also seemed to positively influence the MEP/DOXP and MVA pathways. These data show the most comprehensive metabolomic profile of Cunila menthoides to date, and the effects of BA and GA3 on the synthesis of secondary metabolites, modulating quantitative aspects of metabolism in Lamiaceae.

Variation of biochemical and antioxidant activity with respect to the phenological stage of Tithonia diversifolia Hemsl. (Asteraceae) populations

Tithonia diversifolia is a medicinal plant traditionally used in the treatment of diabetes, diarrhea, menstrual cramps, malaria, hematomas, hepatitis, hepatomas and wounds. This work investigated the influence of environmental factors, phenological stage and genetic variability on the secondary metabolite profile and its antioxidant actions. Leaves were collected at the vegetative stage and in the reproductive stage, in five regions of southeastern Brazil. Phytochemical analyses, antioxidant activity and genetic markers as RAPD (Random Amplified Polymorphic DNA) were used to evaluate the variability between populations and to generate clusters. The highest contents of phenols, flavonoids and tannins were observed in ST (Santa Teresa) samples, as well as displayed great antioxidant activity. ST showed low genetic similarity to other populations (0.12), indicating high levels of genetic diversity and that clustering it is not positively correlated with the proximity of geographical locations. This analysis indicates that phytochemical and antioxidant variations between populations are influenced by genetic factors as well as environmental conditions.

Secondary metabolites profiles and antioxidant activities of germinated brown and red rice

The research aims to investigate the effect of germination on the secondary metabolite profiles and antioxidant activity of brown and red rice. The germination was performed by using a simple laboratory-scale machine that was designed and optimized to provide conditions that support the germination process. The germination was carried out for 2 days in dark conditions at 26°C and 99% humidity. Analysis of the secondary metabolite profile of ungerminated and germinated rice was performed using LC-MS. The antioxidant activities of ungerminated and germinated rice were done by using DPPH method. The results showed that the profiles of secondary metabolites of brown and red rice changed after germination. Some peaks were found to be induced in the germinated rice. However, some peaks were also loss during germination. The antioxidant activity of brown rice was slightly increased due to the germination, from 11.2% to 22.5%. Meanwhile the antioxidant activity of red rice was decreased after germination, from 73.8% to 60.0%.

Targeted and Untargeted Metabolite Profiling of the Ethnobotanical Martynia annua L. Identifies Bioactive Compounds with Medicinal Properties

Abstract Martynia annua is an indigenous plant found endemic in the Indian subcontinent where it is commonly used for self-treatment of a variety of ailments and diseases. Previous studies have shown that preparations from the plant have antiseptic, anti-inflammatory, and antiepileptic effects in vitro, but the biochemical composition and active compounds responsible for health benefits remain unknown. The aim of this study was to take extracts from different parts of the M. annua plant and, using targeted and untargeted metabolite profiling and quantitation, provide a comprehensive secondary metabolite profile to identify potential biotherapeutic metabolites.Ultrahigh-performance liquid chromatography, coupled directly to a Thermo Q-Exactive Orbitrap mass spectrometer, was used for high-resolution targeted and untargeted analysis. 89 metabolites were identified and their relative and selected absolute abundances measured across 5 different parts of the plant (leaf, flower, fruit, stem, and root). A number of the compounds identified are known to have bioactive and therapeutic properties; these include trans-ferulic acid, homovanillic acid, syringic acid, isorhamnetin, apigenin, luteolin, and hispidulin. We report their concentrations in different parts of the plant.Our findings significantly extend the plant metabolite profile of M. annua and amongst the compounds identified, we have highlight those with known biotherapeutic properties. These results provide a foundation for future studies addressing specific compounds that may be responsible for the bioactivity and therapeutic use of M. annua.

The effect of Psephodiplosis rubi (Diptera: Cecidomyiidae) leaf galls on the secondary metabolite profiles of two congeneric host plants

AbstractGalling insects have the unique ability to control the secondary chemistry of their host plants. Galled plant tissue often has higher concentrations of these secondary compounds compared with surrounding plant material. In this study, we used nuclear magnetic resonance (NMR) spectroscopy metabolomics techniques to investigate the effect of galls induced by Psephodiplosis rubi Kolesik, on the profiles of secondary metabolites in its two Rubus host plants in subtropical rainforest. We isolated four triterpenoid compounds from both Rubus species. Concentrations of individual triterpenes were not significantly different in galled and non‐galled leaves; however, the concentration of 2‐oxopolomic acid was significantly negatively correlated with gall abundance. At the interspecific level, non‐galled leaves from both Rubus species were chemically indistinguishable in principal component analysis analyses, while galled leaves showed slight chemical differences. Surprisingly, there was very little difference in metabolic profiles of galled and non‐galled leaves at the intraspecific level. Using NMR, we were able to detect slightly different chemical responses in two congeneric host plant species induced by P. rubi galls. We demonstrate how NMR can be used to detect interspecific chemical differences of plants in ecological studies.

Stachybotrychromenes A\u2013C: novel cytotoxic meroterpenoids from Stachybotrys sp.

In the course of gaining new insights into the secondary metabolite profile of various Stachybotrys strains, in particular concerning triprenyl phenol-like compounds, so far, unknown metabolites with analogous structural features were discovered. Three novel meroterpenoids containing a chromene ring moiety, namely stachybotrychromenes A–C, were isolated from solid culture of the filamentous fungus Stachybotrys chartarum DSMZ 12880 (chemotype S). Their structures were elucidated by means of comprehensive spectroscopic analysis (1D and 2D NMR, ESI-HRMS, and CD) as well as by comparison with spectroscopic data of structural analogues described in literature. Stachybotrychromenes A and B exhibited moderate cytotoxic effects on HepG2 cells after 24 h with corresponding IC50 values of 73.7 and 28.2 μM, respectively. Stachybotrychromene C showed no significant cytotoxic activity up to 100 μM. Moreover, it is noteworthy that stachybotrychromenes A–C are produced not only by S. chartarum chemotype S but also S. chartarum chemotype A and Stachybotrys chlorohalonata.

Nigericin and grisorixin methyl ester from the Algerian soil-living Streptomyces youssoufiensis SF10 strain: a computational study on their epimeric structures and evaluation of glioblastoma stem cells growth inhibition

The present work describes the metabolites produced by a strain identified as Streptomyces youssoufiensis, whose secondary metabolites profile has not been studied so far. The crude ethyl acetate extract was analyzed by high performance liquid chromatography-electrospray ionization mass spectrometry, leading to the detection of the ionophoric polyethers nigericin, epinigericin, abierixin and the newly isolated grisorixin methyl ester. The presence of epimeric forms of nigericin/epinigericin and grisorixin/epigrisorixin has spurred density functional theory computational calculations. This analysis was able to provide the relative stability of the most favored epimers, setting the basis for general structural considerations applicable to several other polyethers. Both nigericin sodium salt and grisorixin methyl ester showed to affect glioblastoma stem cells proliferation in a dose-dependent manner, with a higher activity for the more lipophilic grisorixin methyl ester (GI50 values of 3.85 and 3.05 μM for VIPI and COMI human glioblastoma stem cells, respectively).

The intrinsic quality of brassicaceous vegetables: How secondary plant metabolites are affected by genetic, environmental, and agronomic factors

Abstract From the order Brassicales, different plant organs, such as inflorescences (e.g. broccoli and cauliflower), leaves (e.g. kale and pak choi), heads (e.g. white and red cabbage), as well as roots and bulbs (e.g. radish and turnip), are frequently consumed brassicaceaous vegetables. The order Brassicales is characterized by a specific group of secondary plant metabolites, namely the glucosinolates. Glucosinolates and their breakdown products the isothiocyanates are linked to conferring beneficial health effects. In addition, some studies have also highlighted the beneficial health effects of phenolic compounds and carotenoids, both well-known as antioxidants. Of interest is that the profiles and concentrations of secondary plant metabolites vary enormously between the species, and genetic factors are thought to affect this the most. Further, environmental and agronomical factors are also known to change concentrations of secondary plant metabolites enormously. The main physiological mechanism to produce secondary plant metabolites is defense. Thus, the intrinsic quality, including color, aroma, taste, and beneficial health properties of brassicaceous vegetables, is remarkably affected by secondary plant metabolite profiles and concentrations.

Effects of Developmental Stages and Reduced UVB and Low UV Conditions on Plant Secondary Metabolite Profiles in Pak Choi (Brassica rapa subsp. chinensis).

Pak choi (Brassica rapa subsp. chinensis) is rich in secondary metabolites and contains numerous antioxidants, including flavonoids; hydroxycinnamic acids; carotenoids; chlorophylls; and glucosinolates, which can be hydrolyzed to epithionitriles, nitriles, or isothiocyanates. Here, we investigate the effect of reduced exposure to ultraviolet B (UVB) and UV (UVA and UVB) light at four different developmental stages of pak choi. We found that both the plant morphology and secondary metabolite profiles were affected by reduced exposure to UVB and UV, depending on the plant's developmental stage. In detail, mature 15- and 30-leaf plants had higher concentrations of flavonoids, hydroxycinnamic acids, carotenoids, and chlorophylls, whereas sprouts contained high concentrations of glucosinolates and their hydrolysis products. Dry weights and leaf areas increased as a result of reduced UVB and low UV. For the flavonoids and hydroxycinnamic acids in 30-leaf plants, less complex compounds were favored, for example, sinapic acid acylated kaempferol triglycoside instead of the corresponding tetraglycoside. Moreover, also in 30-leaf plants, zeaxanthin, a carotenoid linked to protection during photosynthesis, was increased under low UV conditions. Interestingly, most glucosinolates were not affected by reduced UVB and low UV conditions. However, this study underlines the importance of 4-(methylsulfinyl)butyl glucosinolate in response to UVA and UVB exposure. Further, reduced UVB and low UV conditions resulted in higher concentrations of glucosinolate-derived nitriles. In conclusion, exposure to low doses of UVB and UV from the early to late developmental stages did not result in overall lower concentrations of plant secondary metabolites.

Chemical profiling, volatile oil analysis and anticholinesterase activity of Hydrocotyle umbellata L. aerial parts cultivated in Egypt

Hydrocotyle umbellata L. (commonly known as Acaricoba) is an aquatic perennial plant belonging to family Araliaceae. The plant has been used in folk phytotherapy and in the Ayurvedic medicine to reduce anxiety and as a memory enhancer herbal remedy. This study was designated to explore the diversity of the accumulated bioactive metabolites that might account for the potential cholinesterase inhibition activity of H. umbellata L. aerial parts. UPLC/PDA/ESI-qTOF-MS secondary metabolites profiling revealed similar chemical composition of the leaves and flowers. The major identified secondary metabolites in both samples were caffeoylquinic acid isomers, quercetin derivatives, and fatty acids. Also, the volatile oil analysis of fresh leaves and flowers using GC/MS showed similar composition. The monoterpenes; α-pinene, β-pinene, and limonene represent the main components of the oil. Many reports were found dealing with the cholinesterase inhibition activity of the major identified caffeoylquinic acids, quercetin derivatives, as well as the identified volatile monoterpenes. Therefore, the anticholinesterase activity of the ethanolic extract of the aerial parts was evaluated using Ellman's spectrophotometric method. The ethanolic extract exhibited significant cholinesterase inhibitory activity with IC50 1.88mg/ml±0.19 compared to standard cholinesterase inhibitor eserine (IC50 0.27mg/ml±0.15). These findings suggest that the significant cholinesterase inhibition activity of the ethanolic extract is likely to be mediated by the major identified bioactive molecules and support the folkloric use of Hydrocotyle umbellata L. as a memory enhancer herbal medicine.