Isolation Of Secondary Metabolites Research Articles

Isolation of secondary metabolites from Geranium molle L. with anticancer potential

Two flavone glycosides, apigenin-6-C-glucoside (1) and lutelin-6-C-glucoside (2), and an anomeric mixture of 2,3-dygalloyl-4,6-HHDP-d-glucose (3a/3b) were isolated for the first time from Geranium molle L., a medicinal plant used in the folk medicine of northeast of Portugal for the treatment of a variety of ailments, including cancer. The cytotoxic activity of the isolated compounds was assessed against several human tumour cell lines and non-tumour porcine liver primary cells. The anomeric mixture of ellagitannins 3a/3b, with the two anomers showing considerably different hydrophobicities, was found to be significantly cytotoxic against cancer cells, with only moderate hepatotoxicity.

Recent metabolomics and gene editing approaches for synthesis of microbial secondary metabolites for drug discovery and development.

Microbial secondary metabolites (SMs) have been identified as an important natural source of drugs for several metabolic and neurological diseases. Along with biomedical applications, SMs are also used in the food and biochemical industries. SMs include natural products such as pigments, alkaloids, toxins, antimicrobials obtained from cultured microorganisms, while other non-cultivable microorganisms have also acted as a rich source of SMs. But, the isolation of SMs from these sources is a very tedious task. Metabolomics provides complete identification and structural information about the entire cellular metabolome under specific conditions using highly sophisticated instrumentation. Further, gene editing techniques such as cloning and gene refactoring, including advanced CRISPR-Cas, can be used for engineering microbes that have the potential to produce natural SMs that were not produced in native microbial strain. The present review describes integrated metabolomics and gene editing approaches for the synthesis of novel microbial SMs and their potential application towards drug discovery and development.

The Phylum Bryozoa as a Promising Source of Anticancer Drugs.

Recent advances in sampling and novel techniques in drug synthesis and isolation have promoted the discovery of anticancer agents from marine organisms to combat this major threat to public health worldwide. Bryozoans, which are filter-feeding, aquatic invertebrates often characterized by a calcified skeleton, are an excellent source of pharmacologically interesting compounds including well-known chemical classes such as alkaloids and polyketides. This review covers the literature for secondary metabolites isolated from marine cheilostome and ctenostome bryozoans that have shown potential as cancer drugs. Moreover, we highlight examples such as bryostatins, the most known class of marine-derived compounds from this animal phylum, which are advancing through anticancer clinical trials due to their low toxicity and antineoplastic activity. The bryozoan antitumor compounds discovered until now show a wide range of chemical diversity and biological activities. Therefore, more research focusing on the isolation of secondary metabolites with potential anticancer properties from bryozoans and other overlooked taxa covering wider geographic areas is needed for an efficient bioprospecting of natural products.

Lantana camara: Secondary Metabolite Isolation by Analytical Techniques

The Plant Lantana camara belongs to the family Verbenaceae, have always been an important source of phytomedicinal agents since ancient times, Until today, it continue to provide modern medicine with novel treatments and to support to identify and isolate compounds from Indian flora with potential biological activity and medicinal value. It has been reported to be used in folk remedies For instance, used for antibacterial, antiulcer, antioxidant, and also treatment for malaria, rheumatisms, asthma, tumors. Many Literature review and phytochemical investigations have been done on this plant, reported to contain various compounds like triterpenoids, proteins, carbohydrates, lactones, furfural, flavonoids, alkaloids, glycosides, tannins, steroids. The ethanolic extract were subjected for column chromatography for the isolation of secondary metabolites by using stationary phase as silica gel with mesh number of 230-400 and the mobile phase was 20% & 30% ethyl acetate/hexane. The Functional groups, structural analysis of the isolated metabolites identified from IR spectrum resembled functional groups of flavonoid chemical structure, Yellow color is characteristic of flavonoids. Keywords: Lantana camara, secondary metabolites, Column chromatography, TLC, IR spectroscopy.

Antioxidant and cytotoxic activity of three Turkish marine-derived fungi

Abstract Background Marine-derived fungi are appraised as a favorable source for discovering new bioactive secondary metabolites. In the last few decades researchers have concentrated on marine-derived fungi to obtain new and pharmaceutically active bioactive secondary metabolites with therapeutic potential. Objective In this study three marine-derived fungi were isolated and identified from marine invertebrates and investigated with regard to their antioxidant and cytotoxic activities. Materials and methods DPPH, SO, NO, and ABTS assays were used for monitoring free radical scavenging activity, and the MTT assay was used for testing cytotoxic activity against HCT-116 colon cancer cells. Results According to the obtained results Malassezia restricta extract was shown to have the highest antioxidant and cytotoxic activities compared to the other tested fungi strains. Conclusion This study is the first report about the antioxidant and cytotoxicity activity of Acremonium sclerotigenum, Aspergillus flavus, and M. restricta. This serves as a valuable preliminary study for activity-guided isolation of secondary metabolites.

Anti-inflammatory effects of Lycium barbarum leaf extracts in lipopolysaccharide-induced RAW 264.7 macrophage cells and isolation of secondary metabolites

Anti-inflammatory effects of Lycium barbarum leaf extracts in lipopolysaccharide-induced RAW 264.7 macrophage cells and isolation of secondary metabolites

Influence of exogenous cinnamic acid on the production of chlorogenic acid in Cichorium intybus L cell culture

Cichorium intybusL(C. intybus) is a source for several important secondary metabolites (SM), including 3-O-caffeoylquinic acid (CGA). Isolation of SM from field plants is difficult due to their batch to batch variation in response to various environmental factors. The study aims to circumvent this problem in C. intybus by increasing CGA concentration through optimization of media conditions and elicitors using plant cell culture. An increased callus growth was observed in leaf explant inoculated on MS solid medium supplemented with 2 mg/l 1-napthaleneacetic acid (NAA), 2 mg/l kinetin (Kn), 30 g/l sucrose and 0.7% agar. Optimum callus growth (13 g/l) was obtained with 5% callus (w/v) sub-cultured in MS suspension medium containing 2 mg/l NAA, 2 mg/l Kn, and 50 g/l glucose at pH 5.5. After optimization of culture conditions, elicitors (cinnamic acid, jasmonic acid, chitosan and methyl jasmonate) were added towards the end of log phase to increase CGA content in callus. Among the various concentrations of elicitors, 0.5 mg/l cinnamic acid was found to reduce cell death and increase phenol content in callus as well as the culture media. These results correlated with the TLC-densitometry study, which showed cinnamic acid (0.5 mg/l) treatment to increase CGA content in callus by 0.39- fold in comparison to control callus. The elicitation was confirmed when 3T3-L1 adipocytes treated with 1 ng/ml methanol extract of callus exposed to 0.5 mg/l cinnamic acid showed a significant 0.3-fold increase in glucose uptake compared to 1 ng/ml control callus and field plant methanol extracts. The elicited callus showed a decrease in the levels of hydrogen peroxide, nitric oxide, and phenylalanine ammonia lyase activity, demonstrating the role of cinnamic acid as a precursor for CGA synthesis.

Terpenes from Zingiber montanum and Their Screening against Multi-Drug Resistant and Methicillin Resistant Staphylococcus aureus

Bioassay directed isolation of secondary metabolites from the rhizomes of Zingiber montanum (Fam. Zingiberaceae) led to the isolation of mono-, sesqui-, and di-terpenes. The compounds were characterized as (E)-8(17),12-labdadiene-15,16-dial (1), zerumbol (2), zerumbone (3), buddledone A (4), furanodienone (5), germacrone (6), borneol (7), and camphor (8) by analysing one-dimensional (1D) (1H and 13C) and two-dimensional (2D) (COSY, HSQC, HMBC, and NOESY) NMR data and mass spectra. Among these terpenes, compounds 1 and 2 revealed potential antibacterial activity (minimum inhibitory concentrations (MIC) values 32–128 µg/mL; 0.145–0.291 mM)) against a series of clinical isolates of multi-drug resistant (MDR) and Methicillin resistant Staphylococcus aureus (MRSA).

Phytochemistry of Three Ecuadorian Lamiaceae: Lepechinia heteromorpha (Briq.) Epling, Lepechinia radula (Benth.) Epling and Lepechinia paniculata (Kunth) Epling.

In this research, the leaves of Lepechinia heteromorpha (Briq.) Epling, Lepechinia radula (Benth.) Epling and Lepechinia paniculata (Kunth) Epling have been collected in order to perform a phytochemical study. The first species was distilled to obtain a novel essential oil (EO), while the others were submitted to ethyl acetate extraction and secondary metabolite isolation. The chemical composition of the EO from L. heteromorpha has been investigated by Gas Chromatography-Mass Spectrometry (GC-MS) and Gas Chromatography with Retention Indices (GC(RI)), identifying 25 constituents. A major compound, (−)-ledol (21.2%), and a minor compound, (−)-caryophyllene oxide (1.0%), were isolated from the EO and their structures confirmed by Nuclear Magnetic Resonance (NMR) spectroscopy. Other major constituents of the EO were viridiflorene (27.3%), (E,E)-α-farnesene (1.4%), spirolepechinene and (E)-β-caryophyllene (7.1% each), allo-aromadendrene (6.1%), camphor (1.7%), limonene (1.3%) and β-phellandrene (4.6%). The enantiomeric composition of the EO monoterpene fraction was also studied, determining the enantiomeric excess and distribution of α-pinene, limonene, β-phellandrene and camphor. The ethyl acetate extract of L. radula and L. paniculata were fractionated by column chromatography. Spathulenol, angustanoic acid E and 5-hydroxy-4′,7-dimethoxy flavone were isolated from L. radula extract; ledol, guaiol and (−)-carnosol were found in L. paniculata.

Effects of sodium nitroprusside on callus browning of Ficus religiosa: an important medicinal plant

Tissue browning is a major problem in tissue culturing of woody plants, especially for Ficus religiosa which occurs by the accumulation and oxidation of phenolic compounds. This study aimed to determine the effect of different concentrations of sodium nitroprusside on the appearance of callus browning from leaf explants. The results indicate that callus browning was significantly reduced by supplementation of sodium nitroprusside to the MS medium and supplemented with 2.26 μM of 2,4-dichlorophenoxyacetic acid and 0.22 μM of 6-benzyl amino purine. The accumulation of hydrogen peroxide and phenolic compounds in the callus tissues decreased at the 50 μM concentration of sodium nitroprusside. Although catalase and peroxidase activities decreased at the 50 μM concentration, the activity of superoxide dismutase and polyphenol oxidases, as well as proline content, increased exponentially. Sodium nitroprusside could be useful for the formation of non-embryogenic callus with high levels of metabolic activity for the production and isolation of secondary metabolites.

Daldinia sacchari (Hypoxylaceae) from India produces the new cytochalasins Saccalasins A and B and belongs to the D. eschscholtzii species complex

Stromata of a Daldinia species were collected from half-burnt sugarcane stalks in South India. Based on a combination of morphological and chemotaxonomic evidence, the species was identified as the first recent record of Daldinia sacchari. While it was impossible to obtain cultures from the ascospores, the stromata were subjected to DNA extraction and DNA sequencing and secondary metabolites were analyzed by high-performance liquid chromatography coupled with diode array and mass spectrometric detection (HPLC-DAD/MS). The fruiting body extract was subjected to preparative HPLC for the isolation of secondary metabolites. Two new cytochalasins, for which we propose the trivial names saccalasins A and B, were elucidated besides two known cytochalasins and binaphthalene tetrol (BNT) by NMR spectroscopy and mass spectrometry. Whereas sequencing of housekeeping genes from the stromatal DNA unfortunately failed, an ITS DNA sequence was obtained from this species for the first time and compared to those of related Hypoxylaceae in a phylogenetic tree. The results indicate a close relationship of D. sacchari to the D. eschscholtzii complex, from which cytochalasins are also known as predominant stromatal metabolites. Phylogenetic analyses based on the ITS-rDNA barcode (which can only discriminate the Hypoxylaceae and other Sordariomycetes into species groups, rather than serve as a means of genus or species discrimination) confirmed that D. sacchari belongs to the D. eschscholtzii species complex. However, as with the majority of tropical Hypoxylaceae species, the availability of cultures that can be used to generate DNA sequence data that are more conclusive than ITS will be imperative to further narrow down the phylogenetic affinities of these fungi.

New secondary metabolites produced by the phytopathogenic fungus Wilsonomyces carpophilus

Two new metabolites possessing the unusual 1-oxa-7-azaspiro[4.4]non-2- ene-4,6-dione core (2, 3) along with the recently described pseurotin A3 (1) were isolated from the pathogenic fungus Wilsonomyces carpophilus(previously named Stigmina carpophila). The producer organism was obtained from Prunus armeniaca collected in Iran and was identified by morphological and molecular phylogenetic methods. The structures of the isolated compounds were elucidated on the basis of extensive NMR spectroscopic analysis, high-resolution mass spectrometry and ECD analysis. The compounds were screened for their antimicrobial, cytotoxic, nematicidal and biofilm inhibition activities but, no significant effect was observed. To the best of our knowledge, this is the first report on the isolation of secondary metabolites produced by W. carpophilus.

Isolation of secondary metabolites from Stenochlaena palustris stems and structure-activity relationships of 20-hydroxyecdysone derivatives on antitermite activity

AbstractStenochlaena palustrisis an edible fern, belonging to the Blechnaceae family and is called kelakai in South Kalimantan, Indonesia. The effects of fractions ofS. palustrisstems and isolated constituents on termite mortality and paper disc consumption byCoptotermes curvignathushave been studied. Treatment of the ethyl acetate (EtOAc) fraction on paper discs greatly induced death ofC. curvignathuswithin 6 days. Our phytochemical investigation ofS. palustrisstems led to the isolation of major constituents and 20-hydroxyecdysone exerted the highest termiticidal activity, followed by stenopalustroside A and ajugasterone C. Moreover, the structure-activity relationships (SAR) of synthetic derivatives from 20-hydroxyecdysone and ajugasterone C suggested that a 2,3-diol has considerable effect on their antitermite properties againstC. curvignathus.

Isolation of Secondary Metabolites with Antimicrobial Activities from Bacillus amyloliquefaciens LWYZ003

The strain LWYZ003, which can restrain multiple pathogens, was screened from the sediment of the ocean and identified as Bacillus amyloliquefaciens. Large-scale fermentation and modern chromatographic separation technologies (macroporous resin column chromatography, silica gel column chromatography, thin-layer chromatography and high-performance liquid chromatography) were used to separate antimicrobial products from the fermentation broth of marine-derived Bacillus amyloliquefaciens LWYZ003. Bioactive-guided separation was used in the term of seeking antimicrobial products from the secondary metabolites of Bacillus amyloliquefaciens LWYZ003. As a result, two natural products cycloheximide (1) and trehalose (2) were obtained. Their structures were elucidated by Fourier transform infrared spectroscopy, high-resolution mass spectrometry, 1H and 13C nuclear magnetic resonance analysis. In the cylinder plate method, compound 1 exhibited stronger antimicrobial activities than compound 2 against Micrococcus luteus, and also exhibited wider antimicrobial spectrum than compound 2. In conclusion, isolation of bioactive secondary metabolites from marine Bacillus sp. has enormous potentials in finding suitable antibiotics to inhibit multiple pathogens.

Characterization of New Bioactive Enzyme Inhibitors from Endophytic Bacillus amyloliquefaciens RWL-1.

Endophytic bacteria are known to produce a wide array of bioactive secondary metabolites with beneficial effects on human health. In the current study, a novel endophytic bacterial strain, Bacillus amyloliquefaciens RWL-1, was isolated from the seeds of Oryza sativa. Initially, the crude extract of RWL-1 was assessed for potential biological effects of enzyme inhibition and cytotoxicity and was found to exhibit a broad spectrum inhibition for α-glucosidase (37 ± 0.09%) and urease (49.4 ± 0.53%). The screening results were followed by bioassay-guided isolation of secondary metabolite(s) from RWL-1. Extensive chromatographic and spectrophotometry analyses revealed the presence of compound 1 (S)-2-hydroxy-N-((S)-1-((S)-8-hydroxy-1-oxoisochroman-3-yl)-3-methylbutyl)-2-((S)-5-oxo-2,5-dihydrofuran-2-yl)acetamide. Further bioassays of compound 1 showed significant inhibition of α-glucosidase (52.98 ± 0.8%) and urease (51.27 ± 1.0%), compared with positive control values of 79.14 ± 1.9% and 88.24 ± 2.2%, and negative controls (0.08 ± 0.1% and 0.05 ± 0.01%), respectively. The current study suggests that bacterial endophytes are a rich source of novel bioactive compounds with high therapeutic value.

Griseofulvin, Diketopiperazines and Cytochalasins from Endophytic Fungi Colletotrichum crassipes and Xylaria sp., and Their Antifungal, Antioxidant and Anticholinesterase Activities

One new natural product, 1-phenylethyl-O-a-L-rhamnopyranoside, together with eight known diketopiperazines and two benzene derivatives were isolated from the endophytic fungus Colletotrichum crassipes, whereas six known compounds including two griseofulvins, three cytochalasins and one dihydroisocoumarin were produced from the endophyte Xylaria sp., both endophytic fungi are associated with leaves of Casearia sylvestris. Cyclo(D)-Pro-(D)-Phe, N-(2-phenylethyl)acetamide and 5-carboxy-6-hydroxy-3-methyl-3,4-dihydroisocoumarin exhibited potent antifungal activity against Cladosporium cladosporioides and C. sphaerospermum. The compounds cyclo(D)-Pro-(D)-Phe, griseofulvin, cytochalasin B and cytochalasin D exhibited antioxidant activity, and the compounds cyclo(D)-Pro-(D)-Phe, cytochalasin B and cytochalasin C showed potent acetylcholinesterase (AChE) inhibition. In addition, this is the first report on the isolation of secondary metabolites from two endophytic fungal strains, associated with Casearia sylvestris.

Isolation of Secondary Metabolites from the Roots of Salix Babylonica

Chemical examination of ethanolic extract of Salix babylonica, root, led to isolation of 2-(Hydroxymethyl) phenyl β-D-glucopyranoside(1) and 2-[(benzoyloxy)methyl]-4-hydroxyphenyl β-D-glucopyranoside 3-Benzoate (2)with previously known compounds β-Sitosterol, kaempferol 7-O-glucoside andapigenin 7-O-galactoside. Compounds 1and 2 had been isolated for the first time from this plant. The structures of the compounds had been elucidated with the help of spectral and chemical studies.

Biophysical and biochemical characterization of active secondary metabolites from Aspergillus allahabadii

Fungal secondary metabolites are a diverse group of natural chemical products with physiological relevance. We aimed to identify bioactive secondary metabolites from Aspergillus allahabadii. We used “activity-guided fractionation” strategy for the isolation of secondary metabolites. Crude extracts showed good antibacterial activity. Two antibacterial secondary metabolites have been isolated from the crude extract. Chemical characterization of these compounds was performed using biophysical techniques (FT-IR, NMR, and mass spectrometry). Structural characterization confirmed these to be pyrone derivatives: 3-hydroxy 2-methyl 4-pyrone and 5-hydroxy-2-(hydroxymethyl)-4H-pyrone. These bioactive pyrone derivatives have been identified as maltol and kojic acid. From our initial observations, we infer that these pyrone derivatives have potent antimicrobial, antioxidant, antidiabetic, and mosquito larvicidal activities and no cytotoxicity. These compounds could have potential therapeutic and biomedical applications, but further mechanistic studies using animal models are very much necessary.

New Enzyme-Inhibitory Triterpenoid from Marine Macro Brown Alga Padina boergesenii Allender & Kraft.

In continuation to our study of the chemical and biological potential of the secondary metabolites isolated from Omani seaweeds, we investigated a marine brown alga, Padina boergesenii. The phytochemical investigation resulted in the isolation of a new secondary metabolite, padinolic acid (1), along with some other semi-pure fractions and sub-fractions. The planar structure was confirmed through MS and NMR (1D and 2D) spectral data. The NOESY experiments coupled with the biogenetic consideration were helpful in assigning the stereochemistry in the molecule. Compound 1 was subjected to enzyme inhibition studies using urease, lipid peroxidase, and alpha-glucosidase enzymes. Compound 1 showed low to moderate α-glucosidase and urease enzyme inhibition, respectively, and moderate anti-lipid peroxidation activities. The current study indicates the potential of this seaweed and provides the basis for further investigation.

Phytochemical and Biological Investigations of Eurya acuminata (Theaceae)

The methanol extract of the powdered leaf of Eurya acuminata was investigated for isolation of secondary metabolites and two compounds were obtained by using VLC, column chromatography and TLC. The compounds were identified as phytol (1) and ?-sitosterol by extensive spectroscopic studies, including high field NMR analyses as well as co-TLC with authentic samples. The methanol extract of leaf of E. acuminata and its organic and aqueous soluble partitioning materials were evaluated for cytotoxic, thrombolytic and antimicrobial properties. In the cytotoxicity study the aqueous fraction of crude methanolic extract showed significant lethality towards brine shrimp having LC50 value 8.821 ?g/ml as compared to standard vincristine sulfate (0.404 ?g/ml). In the study for thrombolytic property, different extract of E. acuminata exhibited various thrombolytic activity ranging from 13.66 to 31.89 % as compared to standard streptokinase (46.51 %). No antimicrobial activity was observed from leaf extracts.Dhaka Univ. J. Pharm. Sci. 15(2): 151-154, 2016 (December)