Isolation Of Secondary Metabolites Research Articles

Bioactivity-Guided Isolation of Secondary Metabolites with Antioxidant and Antimicrobial Activities from Camellia fascicularis.

Camellia fascicularis has important ornamental, medicinal, and food values, which also have tremendous potential for exploiting bioactivities. We performed the bioactivity-guided (antioxidant and antimicrobial) screening of eight fractions obtained from the ethyl acetate phase of C. fascicularis. The antioxidant activity was measured by DPPH, ABTS, and FRAP, and the antibacterial activity was measured by the minimum inhibitory concentration (MIC) of Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus. The results of bioactivity-guided isolation indicated that the major antioxidant compounds in the ethanolic extracts of C. fascicularis may be present in fractions (Fr.) (A-G, obtained after silica gel column chromatography). Fr. (D-I, obtained after silica gel column chromatography) is a fraction of C. fascicularis with antimicrobial activity. The structures of compounds were determined by spectral analysis and nuclear magnetic resonance (NMR) combined with the available literature on secondary metabolites of C. fascicularis leaves. In this study, 17 compounds were identified, including four phenolics (1, 3-4, and 14), a phenylpropane (2), five terpenoids (5-7, 12, and 15), four flavonoids and flavonoid glycosides (8-10 and 16), and two lignins (13 and 17). Compounds 4-7, 13-15, and 17 were isolated from the genus Camellia for first time. The remaining compounds were also isolated from C. fascicularis for first time. The evaluation of antioxidant and antimicrobial activities revealed that compounds 1, 3, 9, 11, and 17 exhibited higher antioxidant activity than the positive control drug (ascorbic acid), and compounds 4, 8, 10, and 13 showed similar activity to ascorbic acid. The other compounds had weaker or no significant antioxidant activities. The MIC of antibacterial activity for compounds 4, 7, and 11-13 against P. aeruginosa was comparable to that of the positive control drug tetracycline at 125 µg/mL, and other secondary metabolites inhibited E. coli and S. aureus at 250-500 µg/mL. This is also the first report of antioxidant and antimicrobial activities of compounds 5-7, 13-15, and 17. The results of the study enriched the variety of secondary metabolites of C. fascicularis and laid the foundation for further research on the pharmacological efficacy and biological activity of this plant.

GC/MS chemical profiling and isolation of secondary metabolites from Leontice ewersmannii

GC/MS chemical profiling of the aerial parts of Leontice ewersmannii revealed the presence of various bioactive compounds of which 20 compounds were in n-hexane fraction and 3 compounds in basic DCM fraction. Among the identified compounds alkaloids, fatty acids and their esters, fatty alcohols, hydrocarbons, and sterol were detected. Phytochemical investigation of the aerial parts of L. ewersmannii led to isolation of 17 compounds including 1-heptacosanol (1), phytol (2), lupenone (3), palmitic acid (4), oleic acid (5), stigmasterol (6), stigmast-7-en-3-ol (7), methyl palmitate (8), methyl linoleate (9), butyl palmitate (10), butyl linoleate (11), dimethyl dodecanedioate (12), methyl cerebronate (13), β-sitosterol (14), β-sitosterol 3-O-β-glucopyranoside (15), sophoridine (16) and camoensine (17). The structures of the isolated compounds were determined by 1D NMR, HR-ESI-MS, and GC-MS. All methyl esters of fatty acids are possible artifacts during extraction. Additionally, the antimicrobial activity of the total extract and different fractions was screened, and they exhibited no inhibition of the growth at the tested concentration ranging from 8 to 200 μg/mL.

Secondary metabolites and essential oil analysis of Clematis simensis with antioxidant and antibacterial activities

Clematis simensis is one of the most commonly used folk medicines in Ethiopia. This study was aimed to investigate the chemical constituents, antioxidant and antibacterial activities of the aerial part of Clematis simensis, and analysis of the composition of its essential oil. The phytochemical study led to the isolation of five compounds: 1-tricosanol (1), 1-hexacosanol (2), 2-deoxy-D-ribono-1,4-lactone (3), 5-hydroxylevulinic acid (4), and β-sitosterol-3-O-β-D-glucoside (5). Their chemical structures were determined by a comprehensive analysis of HR-ESI-MS, FT-IR, UV-Vis, and 1D/2D NMR experiments. The chemical compositions of the essential oils obtained by hydrodistillation from the aerial part of Clematis semenses were determined by GC-MS, which led to the detection of ten major compounds: (E)-2-nonen-1-ol (38.33%), 4-cyclopentene-1,3-dione (28.69%), cyclofenchene (5.65%), benzene acetaldehyde (4.74%), methyl salicylate (2.77%), salicylaldehyde (2.68%), benzaldehyde (2.60%), α-terpineol (2.10%), eucalyptol (1.21%), and 2,6-dimethylcyclohexanol (1.15%). Compound 4 showed high antioxidant activity (IC50 14.89 μg/mL) but has no antibacterial activity against both Gram-positive and Gram-negative bacterial strains. According to the antimicrobial parameters, crude extract showed very weak activity at 500 mg/mL, 250 mg/mL, and 125 mg/mL. To our knowledge, this is the first report on the isolation of secondary metabolites from Clematis simensis. KEY WORDS: Clematis simensis, Essential oil, Phytochemical investigation, Biological activities Bull. Chem. Soc. Ethiop. 2024, 38(4), 1037-1049. DOI: https://dx.doi.org/10.4314/bcse.v38i4.18

Isolation of Secondary Metabolites From Marine Actinobacterium of Microbispora sp.T3S11 and Their Antibacterial Activities.

Introduction Marine actinobacteria are promising sources of novel bioactive compounds due to their distinct ecological niches and diverse secondary metabolite production capabilities. Among these,Microbisporasp. T3S11 is notable for its unique spore chain structure, which allows for both morphological and genetic identification. Despite its potential, little is understood about the secondary metabolites produced by this strain. In this study, we hope to fill this gap by extracting and analyzing the antibacterial activities of secondary metabolites fromMicrobisporasp. T3S11, which will be the first time its bioactive compound profile is investigated. Aim To evaluate the antibacterial activity of secondary metabolites isolated from the marine actinobacteriumMicrobisporasp. T3S11. Materials and methods The antibacterial assays were carried out on agar plates containing the appropriate media for each pathogen. Sterile filter paper disks were impregnated with secondary metabolites extracted fromMicrobisporasp. T3S11 and placed on the surface of agar plates inoculated with the appropriate pathogens. Similarly, disks containing tetracycline were used as a positive control. The plates were then incubated at the appropriate temperature for each pathogen, and the zones of inhibition around the disks were measured to determine the extracted metabolites' antibacterial activity. Result Secondary metabolites had antimicrobial activity againstStreptococcus mutans,Klebsiella pneumonia, and methicillin-resistantStaphylococcus aureus(MRSA). The inhibition ofS. mutanswas 7.5 mm and 8.5 mm at 75 μg/mL and 100 μg/mL, respectively.Klebsiella pneumoniazones measured 7 mm and 7.5 mm, while MRSA zones measured 7.6 mm and 8.5 mm at the same concentrations. Tetracycline, the standard antibiotic, had larger inhibition zones: 22 mm forS. mutansandKlebsiella pneumoniaand 16 mm for MRSA, indicating variable susceptibility. Conclusion We conclude that the secondary metabolites extracted fromMicrobisporasp. T3S11exhibits high antibacterial activity. This could be attributed to the presence of various active compounds.

One-step Bio-guided Isolation of Secondary Metabolites from the Endophytic Fungus Penicillium crustosum Using High-resolution Semi-preparative HPLC.

An endophytic fungal strain Penicillium crustosum was isolated from the seagrass Posidonia oceanica and investigated to identify its antimicrobial constituents and characterize its metabolome composition. The ethyl acetate extract of this fungus exhibited antimicrobial activity against methicillin-resistant Staphylococcus aureus (MRSA) as well as an anti-quorum sensing effect against Pseudomonas aeruginosa. The crude extract was profiled by UHPLC-HRMS/MS, and the dereplication was assisted by feature-based molecular networking. As a result, more than twenty compounds were annotated in this fungus. To rapidly identify the active compounds, the enriched extract was fractionated by semipreparative HPLC-UV applying a chromatographic gradient transfer and dry load sample introduction to maximise resolution. The collected fractions were profiled by 1H-NMR and UHPLC-HRMS. The use of molecular networking-assisted UHPLC-HRMS/MS dereplication allowed preliminary identification of over 20 compounds present in the ethyl acetate extract of P. crustosum. The chromatographic approach significantly accelerated the isolation of the majority of compounds present in the active extract. The one-step fractionation allowed the isolation and identification of eight compounds (1-8). This study led to the unambiguous identification of eight known secondary metabolites as well as the determination of their antibacterial properties.

Bioactive Compounds from an Endophytic Pezicula sp. Showing Antagonistic Effects against the Ash Dieback Pathogen.

A fungal endophyte originating from the Canary Islands was identified as a potent antagonist against the fungal phytopathogen Hymenoscyphus fraxineus, which causes the devastating ash dieback disease. This endophyte was tentatively identified as Pezicula cf. ericae, using molecular barcoding. Isolation of secondary metabolites by preparative high-performance liquid chromatography (HPLC) yielded the known compounds CJ-17,572 (1), mycorrhizin A (3) and cryptosporioptides A-C (4-6), besides a new N-acetylated dihydroxyphenylalanin derivative 2, named peziculastatin. Planar structures were elucidated by NMR and HRMS data, while the relative stereochemistry of 2 was assigned by H,H and C,H coupling constants. The assignment of the unknown stereochemistry of CJ-17,572 (1) was hampered by the broadening of NMR signals. Nevertheless, after semisynthetic conversion of 1 into its methyl derivatives 7 and 8, presumably preventing tautomeric effects, the relative configuration could be assigned, whereas comparison of ECD data to those of related compounds determined the absolute configuration. Metabolites 1 and 3 showed significant antifungal effects in vitro against H. fraxineus. Furthermore, 4-6 exhibited significant dispersive effects on preformed biofilms of S. aureus at concentrations up to 2 µg/mL, while the biofilm formation of C. albicans was also inhibited. Thus, cryptosporioptides might constitute a potential source for the development of novel antibiofilm agents.

Secondary Metabolite of Endophytic Fungi Daldinia eschscholtzii from The Leaves of Syzygium polyanthum

Salam (Syzygium polyanthum) is a plant that is often used by Indonesian people as traditional medicine. The leaves are consumed to treat various diseases. This study reports the endophytic fungi found from S. polyanthum leaves and its secondary metabolites. Endophytic fungi species were determined through morphological and molecular identification. Potato Dextrose Broth medium was used for the cultivation process and ethyl acetate was used as a solvent for the extraction process. Antioxidant tests was carried out by using the DPPH method. Chromatographic techniques were used for the isolation of secondary metabolite and spectroscopic analysis was carried out to identify their chemical structures. The results of morphological and molecular analysis showed that the sample was Daldinia eschscholtzii. The secondary metabolite obtained from this endophytic fungus was idetified as fonsecinone A with good antioxidant activity. The secondary metabolite have the potential to become a source antioxidants through further research.

Cell culture of <i>Scorzonera hispanica</i> L., strain SFR-SH1. Major secondary metabolites

Plant cell cultures are widely used in the world pharmaceutical industry as a raw material for the production of biologically active compounds. Their certain advantages, such as independence from seasonal conditions, diseases and their transmitter, contribute to the development of this area of biotechnology. In addition, it is possible to obtain the required quantity of the desired product with standard quality characteristics. Since the biosynthesis of new compounds does not occur in the intact plant, another advantage of this method arises. It consists in obtaining new substances due to the peculiarities of cell culture biology. The conducted review represents secondary metabolites isolated from the cell culture of Scorzonera hispanica L., strain SFR-SH1, possessing biological activity. In addition, the history of developing the selected strain and schemes for isolation and purification of secondary metabolites are demonstrated. For a clearer discussion, the main secondary metabolites found in cells are broadly divided into three groups. The first group includes compounds typical of most plants: β-sitosterol and its glucoside, caffeic acid methyl ester and oleanic acid. The second group contains syringaresinol monoglucoside, which is relatively rare in plant objects. The third group includes two compounds isolated from natural objects for the first time: sesquiterpene glucoside, scorzoside and neolignan scorzonoside. The results obtained were compared with the available literature data on the chemical composition of native plants of S. hispanica and other Scorzonera genera species. Due to its high biological activity, Syringaresinol monoglycoside attracts more attention than other compounds detected in cultured cells. Long-term cultivation of Monoglycoside leads to an alteration in the physicochemical characteristics of this compound for an unknown reason.

Оптимизация получения полисахаридов из Cyanobacterium sp. IPPAS B-1200

Cyanobacterium sp. IPPAS B-1200 is a cyanobacteria strain that belongs to the rare genus Cyanobacterium, family Cyanobacteriaceae fam. nov. Studies devoted to the isolation of secondary metabolites from this strain concentrate mostly on the fatty acid composition while the issue of isolating and identifying exopolysaccharides remains understudied. However, polysaccharides from cyanobacteria are of scientific and economic interest in the framework of biotechnology, medicine, pharmacology, etc. The research objective was to study the effect of the physicochemical conditions of cultivation and the composition of the cultural medium on exopolysaccharide production.
 Cyanobacterium sp. B-1200 were grown under 7500 ± 50 lux (12 h light/12 h dark). The dry cell weight was determined by gravimetry and a calibration plot that illustrated the dependence of the biomass amount on the degree of absorption at a wavelength of 750 nm. The amount of polysaccharides in the culture liquid was assessed by the Anthrone-sulphate method. The extraction was carried out by alcohol precipitation. The method of ultrasonic dispersion was used to destroy the cell walls of cyanobacteria. 
 The experimental study revealed the optimal parameters for the extraction and purification of exopolysaccharides from the culture medium. Removal of sodium bicarbonate from the medium and a 300%-increase in its concentration raised the yield of polysaccharides. The optimal value of active acidity for the synthesis of polysaccharides was pH = 6 while the optimal temperature for their accumulation was 35°C. The largest amount of biomass was obtained at 25°C. Intense white illumination contributed to the greatest release of exopolysaccharides into the culture medium; red-white illumination affected the morphology of cyanobacteria cells. During the extraction, the concentration, temperature, and nature of the extractant proved to be the most important factors. For example, isopropanol produced the highest yield while butanol triggered the least effective response. 
 The optimal extraction and purification modes for polysaccharides were as follows. For ultrasonic processing, the best results were obtained at a power of 20 W after 5 min. For freeze drying, the rational parameters were 8 h at –15°C.

Guided isolation of secondary metabolites from Nectria sp. MHHJ-3 by molecular network strategy

The fungus Nectria sp. MHHJ-3 was isolated from Illigera rhodantha. A molecular networking-guided the secondary metabolites investigation of Nectria sp. MHHJ-3 led to the isolation of ten metabolites (1−10), including two new naphthalenone derivatives, nectrianaphthalenones A (1) and B (2), and two new steroids, nectriasteroids A (3) and B (4). Their structures were elucidated by extensive spectroscopic analysis including the HRESIMS, 1D/2D NMR and electronic circular dichroism (ECD) spectra. A plausible biosynthetic pathway for 1–2 was proposed. Compounds 1 and 2 exhibited moderate acetylcholinesterase (AChE) inhibitory activities. Compounds 3 and 4 showed significant cytotoxic activity against selected tumor cells. Particularly, compound 3 exhibited the strongest activity against A549 cells with an IC50 value of 13.73 ± 0.03 μM, which was at the same grade with that of positive control cisplatin.

Investigation of Small-Molecule Constituents in Voacanga africana Seeds and Mapping of Their Spatial Distributions Using Laser Ablation Direct Analysis in Real-Time Imaging-Mass Spectrometry (LADI-MS).

Plant seeds are a renewable resource that can furnish access to medicinal natural products that can only otherwise be isolated from aerial or root parts, the harvest of which may be destructive to the plant or threaten its viability. However, optimization of the isolation of such compounds from seeds would be greatly assisted if the spatial distribution of the molecules of interest within the plant tissue were known. For example, iboga alkaloids that hold promise for the treatment of opioid use disorder are typically isolated from the leaves, bark, or roots of Tabernanthe or Voacanga spp. trees, but it would be more environmentally sustainable to isolate such compounds from their seeds. Here, we leveraged the unique capabilities of the ambient mass spectral imaging technique termed laser ablation direct analysis in real-time imaging-mass spectrometry (LADI-MS) to reveal the spatial distributions of a range of molecules, including alkaloids within V. africana seeds. In addition to six compounds previously reported in these seeds, namely, tetradecanoic acid, n-hexadecanoic acid, (Z,Z)-9,12-octadecadienoic acid, (Z)-9-octadecenoic acid, octadecanoic acid, and Δ14-vincamine, an additional 31 compounds were newly identified in V. africana seeds. The compound classes included alkaloids, terpenes, and fatty acids. The ion images showed that the fatty acids were localized in the embryo of the seed. The alkaloids, which were mainly localized in the seed endosperm, included strictamine, akuammidine, polyneruidine, vobasine, and Δ14-vincamine. This information can be exploited to enhance the efficiency of secondary metabolite isolation from V. africana seeds while eliminating the destruction of other plant parts.

Characterization of rhizobacteria secondary metabolites on maize (Zea mays) in marginal land

Maize is one of the strategic food crops and has economic value and could be developed because of its position as the main carbohydrate and protein source after rice. This could be a reference for us to increase maize cultivation, seeing the role of maize is quite large, and the availability of maize is less stable every year. maize can grow optimally on productive land and marginal land, but at this time more productive land is used for rice production. One alternative for maize production is on marginal land especially dryland. The main problem in dryland is limited water. One way to overcome this is by inoculating Rhizobacteria which can increase the growth and resistance of maize to drought. Rhizobacteria possess tremendous potential for modulating the physiological response to water deprivation, thus ensuring plant survival under such stressful conditions. The research aimed to find rhizobacteria have secondary metabolite for resistance to drought. Rhizobacteria exploration and secondary metabolite isolation was conducted at University of Muhammadiyah Malang. The research results have obtained 10 isolates capable of synthesizing extracellular secondary metabolites: osmoprotectants, phytohormones, and organopesticides that can be used by plants. The results of the analysis by GS-MS (Gas Chromatography-Mass Spectrometry) rhizobacteria showed that three compounds were synthesized. The osmoprotectants compound synthesize ware found namely: (a) (2E)-3,7-dimethylocta-2,6-dienal 16.66%;(b) (E)-hept-2-enal12.55%;(c)2-methylsulfanylethanamine 5.34%; (d) 1,3,5-trimethoxybenzene 4.42%. It is also capable of synthesizing phytohormones: (a) cinnolino [4,3-b] quinoxaline 6.00%; (b) 1-methyl-2-phenylindole 5.90%; (c) 1H-Indole 3.64%; (d) N-2-Fluorenylperfluorpropionamid 3.02%. These rhizobacteria were also able to synthesize: (a) 2-chloro-4-methoxy phenol 11.32%; (b) (E)-non-2-enal 9.87%; (c) 4-chloro-2,6-dipyridin-2-ylpyridine 7.07%; (d) 2-(4-chlorophenyl) hexane nitrile 6.45%. The Osmprotectant compounds aim to protect cells from drought conditions, phytohormones to stimulate plant growth and organopesticides to overcome pests and diseases.

Endophytic Fungi from Mangrove Plant Acanthus ilicifolius L.: Antimicrobial, Anticancer, and Species Determination

The current study aimed to screen, identify and evaluate the biological activity of fungi strains from the mangrove plant Acanthus ilicifolius L, which has proven to be a rich source of fungal species. Thirteen fungi were screened from the leaves, stem bark, and roots. Each fungal strain was cultivated in rice as media, and then the secondary metabolites of the fungus were extracted with ethyl acetate. The extracts were tested for antimicrobial and cytotoxic activities. Next, a phylogenetic tree was reconstructed based on the sequence similarities of each strain. The fungal extract AIA1 displayed excellent antimicrobial activity in a concentration of 5 % against some pathogenic bacterial strains with an inhibition zone in the range of 11.87 ± 0.87 to 15.42 ± 0.51 mm. The brine shrimp lethality test (BSLT) showed that strains AIA1 and AIKB3 were toxic against Artemisia salina L with an LC50 value of less than 80 ppm. The MTT assay determined the cytotoxicity against the human ductal breast epithelial tumor (T47D) cell line. Based on molecular identification, AIA1, AIKB3, and AID1 showed maximum similarity (> 95 %) with Pestalotiopsis sp. AIA1, Trichoderma yunnanense AIKB3, and Schizopyllum commune AID1, respectively. The ethyl acetate extracts of fungi strains from the mangrove plant A. ilicifolius showed strong antibacterial and moderate cytotoxic activities, which make them good candidates for secondary metabolite isolation. HIGHLIGHTS This is the first report on exploring the endophytic fungi from the mangrove plant Acanthus ilicifolius collected from Mangrove Forest Tourism Park, West Sumatera, Indonesia Endophytic fungi strains from mangrove plant Acanthus ilicifolius showed good activity on the antimicrobial and cytotoxic assay Three potential endophytic fungi were identified asPestalotiopsis AIA1, Trichoderma yunnanense AIKB3, and Schizopyllum commune AID1 GRAPHICAL ABSTRACT

Isolation of the Anti-Inflammatory Agent Myceliostatin from a Methionine-Enriched Culture of Myceliophthora thermophila ATCC 42464.

Safe and effective nonsteroidal anti-inflammatory drugs are needed. Meanwhile, addition of amino acids to cultures of microorganisms is likely to increase the possibility of novel secondary metabolite isolation. In the course of screening for anti-inflammatory agents using cellular lipopolysaccharide (LPS)-induced nitric oxide (NO) production, two new related compounds with the myceliothermophin structure from a methionine-enriched culture of Myceliophthora thermophila ATCC 42464 were isolated. The new compounds have an additional methylthio group on the myceliothermophin structure and were named myceliostatins A and B. Both compounds inhibited LPS-induced NO production at nontoxic concentrations in macrophage-like mouse monocytic leukemia RAW264.7 cells. Myceliostatin B inhibited the expression of LPS-induced iNOS, IL-6, and IL-1β and the upstream NF-κB activity in situ and in vitro. Finally, it was found to inhibit NF-κB binding to DNA in the reconstruction system with purified p65. Myceliostatin B also inhibited LPS-induced reactive oxygen species (ROS) production. Thus, myceliostatin B, a novel compound derived from M. thermophila, was found to be a new anti-inflammatory and antioxidant compound directly inhibiting NF-κB.

Isolasi Senyawa Metabolit Sekunder Dari Fraksi Aktif Antibakteri Pada Tumbuhan Sirih Merah (Piper porphyrophyllum)

The red betel plant (Piper porphyrophyllum) belongs to the Piperaceae family which is traditionally used to treat various diseases, including headaches, bone pain, chest tightness, leprosy, stomach ache, as well as for various other diseases such as skin diseases and diabetes. In this study, the antibacterial activity was tested for the hexane, ethyl acetate and methanol extracts of red betel nut using the disc diffusion method and the isolation of secondary metabolites from the hexane fraction of red betel nut using column chromatography technique. Determination of the antibacterial activity of the three fractions showed that the methanol extract had better antibacterial activity compared to the hexane extract and ethyl acetate extract. UV spectrum analysis showed 3 bands with absorption of 282 nm in band I, 274.80 nm in band II and 224.80 nm in band III which means that they show characteristics for triterpenoid group compounds. FTIR analysis showed the presence of C-H, CH2, C=O, C-O alcohol and geminal dimethyl functional groups in the isolated compounds.

Could elderberry fruits processed by modern and conventional drying and extraction technology be considered a valuable source of health-promoting compounds?

The influence of drying and extraction processes on the phytochemical composition and biopotential of elderberry is challenging for the food industry. For this reason in this research two drying techniques (lyophilization and natural convection) and three extraction techniques (ultrasound (UAE), microwave (MAE), and conventional (maceration (MAC)) was applied using two”green” solvents (water and 50 % ethanol). Results of the research showed that lyophilization was a better way of drying than natural convection, while MAE extraction was the most efficient technique for the isolation of secondary metabolites. The most abundant phenolic compounds established by LC-MS/MS analysis, were chlorogenic acid and rutin, identified in extracts of lyophilized elderberries. Elderberry extracts achieved great antioxidant (CUPRAC: 2.30–5.13 mg TE/mL) and enzyme inhibitor potential (α-amylase: 0.51–8.34 mg ACAE/mL). The results suggest that elderberry is a rich natural source of bioactive compounds and could be used for the future development of dietary supplements and functional foods.

Targeted Isolation of Anti-inflammatory Lignans from Justicia aequilabris by Molecular Networking Approach.

Herein, the isolation of secondary metabolites from the aerial parts of Justicia aequilabris guided by HPLC-MSn and molecular networking analyses is reported. Twenty-two known compounds were dereplicated. Three new lignans (aequilabrines A-C (1-3)) and three known compounds (lariciresinol-4'-O-β-glucose (4), roseoside (5), and allantoin (6)) were obtained. The anti-inflammatory activity of compounds 1-3 was evaluated in vitro by inhibiting the nitric oxide production (NO) and pro-inflammatory activity on the cytokine IL-1β. Compounds 2 and 3 showed significant inhibitory activity against NO production, with IC50 values of 9.1 and 7.3 μM, respectively. The maximum inhibition of IL-1β production was 23.5% (1), 27.3% (2), and 32.5% (3).

A Systematic Review on Waste as Sustainable Feedstock for Bioactive Molecules—Extraction as Isolation Technology

In today’s linear economy, waste streams, environmental pollution, and social–economic differences are increasing with population growth. The need to develop towards a circular economy is obvious, especially since waste streams are composed of valuable compounds. Waste is a heterogeneous and complex matrix, the selective isolation of, for example, polyphenolic compounds, is challenging due to its energy efficiency and at least partially its selectivity. Extraction is handled as an emerging technology in biorefinery approaches. Conventional solid liquid extraction with organic solvents is hazardous and environmentally unfriendly. New extraction methods and green solvents open a wider scope of applications. This research focuses on the question of whether these methods and solvents are suitable to replace their organic counterparts and on the definition of parameters to optimize the processes. This review deals with the process development of agro-food industrial waste streams for biorefineries. It gives a short overview of the classification of waste streams and focuses on the extraction methods and important process parameters for the isolation of secondary metabolites.

Isolation of secondary metabolite compounds of Coffee Benalu leaves (Loranthus parasiticus (L.) Merr.) and its antibacterial activity test

Coffee parasite leaves (Loranthus parasiticus (L.) Merr) is one of the plants that can be used as a medicinal material commonly obtained in various subtropical regions or tropical regions. This study aims to isolation of secondary metabolite compounds from methanol extract of coffee leaves and antibacterial activity by of disc diffusion and microdilution methods against S. aureus bacteria, S. mutans, and S. viridians. The result of antibacterial activity against S. aureus bacteria, S. mutans, and S. viridians respectively is 7.5 mm; 7.9 mm; and 8 mm which indicates that the ability to inhibit the growth of methanol extract bacteria 1% coffee leaves belongs to the medium category. The results of the microdilution method in determining the value of KHM against bacteria S. Aureus, S. Mutans, and S. Viridians are equal to the KHM value of 5000 μg/mL. Meanwhile, the MBC value for S. aureus was >5000 g/Ml, for S. mutans was >5000 g/mL and for S. viridians it was 5000 g/mL, indicating that the methanol extract of the coffee parasite leaves was only an inhibitor. Isolation of secondary metabolite compounds is carried out by fractionation using Liquid Vacuum Chromatography and Gravitational Column Chromatography which are further characterized using the GC-MS instrument. The results of the isolation of secondary metabolites from the methanol extract of the leaves of the coffee parasite (Loranthus parasiticus (L.) Merr.) showed that the leaves of the coffee parasite contained any 9 compounds.

Cecropia pachystachya Trécul: identification, isolation of secondary metabolites, in silico study of toxicological evaluation and interaction with the enzymes 5-LOX and α-1-antitrypsin

ABSTRACT Cecropia pachystachya Tréc., popularly known as embaúba, belongs to the Cecropiaceae family and is used by the native population in the treatment of bronchitis, asthma, high blood pressure, fever, and as a diuretic. The pharmacological actions including anti-inflammatory, antioxidant, cardiotonic and sedative were previously reported. The objective of this study was to (1) isolate and identify bioactive compounds extracted from the ethanolic extract of C. pachystachya roots (ERCP), as well as (2) verify the affinity of these metabolites with the enzymes 5-lipoxygenase (5-LOX) and α-1-antitrypsin through in silico tests. Isolation and/or identification were performed using GC-MS, HPLC, Infrared (IR), and nuclear magnetic resonance (NMR) techniques. After isolation and identification of the active compounds, these substances were subjected to the in silico investigation that proceeded by performing PreADMET simulations and molecular docking calculations. The bioactive compounds identified were 1-(+)-ascorbic acid 2,6-dihexadecanoate, ethyl hexadecanoate, ethyl (9E,12E)-octadec-9,12-dienoate, ethyl (Z)-octadec-9-enoate and ethyl octadecanoate by GC-MS; chlorogenic acid, catechin, epicatechin, syringaldehyde by HPLC; β-sitosterol, sitostenone, beccaridiol, tormentic acid, lupeol, α- and β-amyrin by classical chromatography, IR, 1H and 13C NMR techniques. The ADMET (absorption, distribution, metabolism, excretion, and toxicity) properties were determined for each bioactive compound. Tormentic acid demonstrated a greater affinity for 5-LOX enzyme while sitostenone demonstrated a higher affinity for the α-1-antitrypsin enzyme. Our findings demonstrated a diverse range of secondary metabolites isolated from C. pachystachya that showed relevant interactions with the enzymes 5-LOX and α-1-antitrypsin. Thus, “embaúba” may be employed in in vivo experimental studies as a form of alternative treatment for chronic lung diseases. Abbreviations: ADT: Autodock Tools; BBB: Blood-brain barrier; CaCo2: Human colonic adenocarcinoma cells; CC: Classic/open Column; TLC: Thin Layer Chromatography; CD40: Differentiation Cluster 40; CENAUREMN: Centro Nordestino de Aplicação e Uso da Ressonância Magnética Nuclear; GC-MS: Gas Chromatography coupled to mass spectrometry; HPLC: High-Perfomance Liquid Chromatography; CYP2C9, CYP2C19, CYP2D6 and CYP3A4: Cytochrome P450 isoenzymes; COPD: Chronic Obstructive Pulmonary Disease; DRX-500: X-Ray Diffraction – 500; ERCP: Ethanolic extract of the roots of C. pachystachya; FAPEPI: Fundação de Amparo à Pesquisa do Piauí; HIA: Human Intestinal Absorption; IR: Infrared; Ki: Inhibition constant; 5-LOX: 5-Lipoxygenase; mM: miliMolar; nM: nanoMolar; OECD423: acute toxic class method; PDB: Protein Data Bank; P-gP: P-glycoprotein; PM2,5: Small inhalable particles 2,5; PPB: Plasm Protein Binding; PreADMET: Prediction Absorption, Distribution, Metabolization, Excretion and Toxicity; NMR: Nuclear Magnetic Resonance; +S9: with metabolic activation; -S9: no metabolic activation; SisGen: Sistema Nacional de Gestão de Patrimônio Genético e do Conhecimento Tradicional Associado; RT: Retention time; TA100: Ames test with TA100 cells line; TA1535: Ames test with cells of the TA1535 cell line; UESPI: State University of Piauí; V79: lung fibroblast cells; ΔG: Gibbs free energy (Kcal/mol); μM: microMolar.