Culture Extracts Research Articles

Antibacterial and Anticancer Properties of Diketopiperazines from Streptomyces antimicrobicus BN122, an Endophyte in Oryza sativa var. glutinosa.

<b>Background and Objectives:</b> This study characterized a bacterial strain, BN122, isolated from the root tissues of purple sticky rice (<i>Oryza sativa</i> L. var. glutinosa). Identified as <i>Streptomyces antimicrobicus</i> based on 16S rDNA analysis and physical-chemical properties, the aim was to isolate and evaluate the antibacterial and anticancer activities of its bioactive compounds. <b>Materials and Methods:</b> The major compounds were purified from BN122's culture extract using column chromatography and TLC. The NMR spectroscopy and mass spectrometry confirmed their identities as Cyclo-(L-Pro-L-Val), Cyclo-(L-Pro-L-Leu), Cyclo-(L-Pro-L-Trp) and Cyclo-(L-Pro-L-Phe). The antibacterial and anticancer activities of these compounds were subsequently assessed. Statistical significance was determined using SPSS software. <b>Results:</b> Isolated compounds exhibited potent antibacterial activity against Gram-positive bacteria. Minimum inhibitory concentrations (MICs) ranged from 32 to 256 μg/mL, while minimum bactericidal concentrations (MBCs) were between 128 and 512 μg/mL. Compounds demonstrated potent cytotoxic activity against cancer cells, with IC<sub>50</sub> values ranging from 32.00 to 57.08 μg/mL for MDA-MB-231 cells, 85.73 to 158.93 μg/mL for HeLa cells and 276.89 to 323.48 μg/mL for HepG2 cells. Notably, these compounds exhibited moderate toxicity towards non-cancerous Vero cells (IC<sub>50</sub> = 482.73 to 680.87 μg/mL). <b>Conclusion:</b> The findings suggested that <i>Streptomyces antimicrobicus</i> BN122 produces compounds with promising antibacterial and anticancer properties. Further research on these compounds could contribute to developing novel therapeutic strategies for bacterial infections and certain cancers.

Advancements of astaxanthin production in Haematococcus pluvialis: Update insight and way forward.

The global market demand for natural astaxanthin (AXT) is growing rapidly owing to its potential human health benefits and diverse industry applications, driven by its safety, unique structure, and special function. Currently, the alga Haematococcus pluvialis (alternative name H. lacustris) has been considered as one of the best large-scale producers of natural AXT. However, the industry's further development faces two main challenges: the limited cultivation areas due to light-dependent AXT accumulation and the low AXT yield coupled with high production costs resulting from complex, time-consuming upstream biomass culture and downstream AXT extraction processes. Therefore, it is urgently to develop novel strategies to improve the AXT production in H. pluvialis to meet industrial demands, which makes its commercialization cost-effective. Although several strategies related to screening excellent target strains, optimizing culture condition for high biomass yield, elucidating the AXT biosynthetic pathway, and exploiting effective inducers for high AXT content have been applied to enhance the AXT production in H. pluvialis, there are still some unsolved and easily ignored perspectives. In this review, firstly, we summarize the structure and function of natural AXT focus on those from the algal H. pluvialis. Secondly, the latest findings regarding the AXT biosynthetic pathway including spatiotemporal specificity, transport, esterification, and storage are updated. Thirdly, we systematically assess enhancement strategies on AXT yield. Fourthly, the regulation mechanisms of AXT accumulation under various stresses are discussed. Finally, the integrated and systematic solutions for improving AXT production are proposed. This review not only fills the existing gap about the AXT accumulation, but also points the way forward for AXT production in H. pluvialis.

Antimicrobial polyketides from the endophytic fungus Fusarium asiaticum QA-6 derived from medicinal plant Artemisia argyi.

Seven previously undescribed polyketide derivatives, fusariumtides A-G (1-7), together with three known analogues (8-10), were isolated from the culture extract of Fusarium asiaticum QA-6, an endophytic fungus obtained from the fresh stem tissue of the medicinal plant Artemisia argyi H. Lev. & Vaniot. Their structures were elucidated by detailed interpretation of 1D/2D NMR spectroscopic and mass spectrometric data, and the absolute configuration of compound 1 was established on the basis of X-ray crystallographic analysis and ECD and specific rotation (SR) calculations. The isolated compounds, which possessed a functionalized decalin moiety, were evaluated for antimicrobial activities. Compounds 1 and 10 exhibited broad-spectrum inhibitory activities against nine tested pathogenic bacteria with MIC values ranging from 1 to 64μg/mL, while compound 8 showed potent inhibitory activities against aquatic pathogens Aeromonas hydrophilia, Pseudomonas aeruginosa, Vibrio harveyi, and V. vulnificus, which were comparable to/or stronger than those of the positive control chloramphenicol.

Preliminary study on the potential damage of cigarette smoke extract in 3D human chondrocyte culture.

Osteoarthritis (OA) is a chronic degenerative disease characterized by the progressive loss of articular cartilage. The role of cigarette smoke (CS) in OA is debated, with some studies suggesting a protective effect while others indicate it may pose a risk. Our preliminary findings suggest a link between smoking in young adults and severe knee OA, though the extent of this contribution is unclear. This study investigates the impact of cigarette smoke extract (CSE) on human chondrocytes. Human chondrocyte cultures were exposed to varying concentrations (0-10%) of CSE for 7 d. We evaluated cell viability, extracellular matrix (ECM) components, metalloproteinaseexpression and cytokines levels, and antioxidant enzymes (SOD1 and CAT) using calcein staining, immunohistochemistry and ELISA. Oxidative stress (OS) was assessed by measuring hydrogen peroxide (H2O2) and nitric oxide (NO) levels. Results were analyzed using ANOVA with Tukey post hoc tests, and Pearson correlation coefficients were calculated. Cell viability decreased at 10% CSE, and ECM components were diminished. MMP9 and MMP13 expression significantly increased at 5% and 10% CSE. H2O2 levels peaked at 1%, while IL-1β peaked at 2.5%. Antioxidant expression (SOD1 and CAT) decreased at higher concentrations, and heat shock protein 70 (HSP70) was notably expressed. MMPs expression was negatively correlated with both viability and ECM components. CSE induces cellular damage, alters ECM composition, and upregulates MMP expression via OS and IL-1β, while diminishing antioxidant defenses. These findings suggest that smoking may disrupt articular cartilage homeostasis, highlighting the need for further investigation into oxidative stress and inflammatory mediators.

Eremophilane- and Acorane-Type Sesquiterpenes from the Deep-Sea Cold-Seep-Derived Fungus Furcasterigmium furcatum CS-280 Cultured in the Presence of Autoclaved Pseudomonas aeruginosa QDIO-4.

Six new sesquiterpenes, including four eremophilane derivatives fureremophilanes A-D (1-4) and two acorane analogues furacoranes A and B (5 and 6), were characterized from the culture extract of the cold-seep derived fungus Furcasterigmium furcatum CS-280 co-cultured with autoclaved Pseudomonas aeruginosa QDIO-4. All the six compounds were highly oxygenated especially 2 and 3 with infrequent epoxyethane and tetrahydrofuran ring systems. The structures of 1-6 were established on the basis of detailed interpretation of 1D and 2D NMR and MS data. Their relative and absolute configurations were assigned by a combination of NOESY and single crystal X-ray crystallographic analysis, and by time-dependent density functional (TDDFT) ECD calculations as well. All compounds were tested the anti-inflammatory activity against human COX-2 protein, among which, compounds 2 and 3 displayed activities with IC50 values 123.00 µM and 93.45 µM, respectively. The interaction mechanism was interpreted by molecular docking.

Localization of Secondary Metabolites in Relict Gymnosperms of the Genus Sequoia In Vivo and in Cell Cultures In Vitro, and the Biological Activity of Their Extracts.

In order to scientifically search for new sources of secondary metabolites with valuable qualities for phytopharmacognosy, tasks requiring a step-by-step solution were set. The primary task is the development of technologies for obtaining in vitro highly productive biomass of cells of relict gymnosperms of the genus Sequoia, capable of accumulating various classes of secondary metabolites. The study of the accumulation and localization of secondary metabolites allowed us to evaluate the biological activity and cytotoxicity of in vitro Sequoia cultures. In our study, histochemical methods were used to determine the localization of secondary compounds (phenolic and terpenoid in nature) in plant tissues. Secondary metabolites-polyphenols, catechins, and terpenoids-are mainly localized in the epidermal, parenchymal, and conductive tissues of Sequoia leaves and stems. In callus and suspension cultures of Sequoia, secondary metabolites were localized in cell walls and vacuoles. The mineral composition of the nutrient medium (MS and WPM), the light source (photoperiod), and the endogenous content of polyphenols in the primary explant influenced the initiation and growth characteristics of the in vitro culture of Sequoia plants. Inhibition of growth in suspension cultures on the WPM nutrient medium was noted. The cultivation of Sequoia cell lines at a 16 h photoperiod stimulated the formation of polyphenols but had a negative effect on the growth of callus cultures. Extractive substances obtained from intact and callus tissues of evergreen Sequoia demonstrate high biological (fungicidal) activity and cytotoxicity. The inhibitory effect on Fusarium oxisporum was noted when 200 mg/L of Sequoia extract was added to the nutrient medium. Extracts of redwood callus cultures were low in toxicity to normal FetMSC cells but inhibited the growth of lines of "immortal" cervical HeLa cancer cells and human glioblastoma A172. Intact tissues of Sequoia plants and cell cultures initiated from them in vitro are producers of secondary metabolites with high biological activity.

Breviane Spiroditerpenoids with Anti-inflammatory and Antiviral Activities from Deep-Sea-Derived Fungus Penicillium sp. F59.

Twelve new breviane spiroditerpenoids, namely, chrysobreviones A-L (1-12), together with seven structurally related analogues (13-19) were isolated from the EtOAc extract of the fermented cultures of deep-sea-derived fungus Penicillium sp. F59. These structures including absolute configurations were resolved on the basis of extensive analysis of NMR spectroscopic data and HRESIMS, in association with experimental and calculated ECD data as well as the modified Mosher's method. Compound 1 represented the first breviane derivative containing an unusual octahydrodifuro[2,3-b:2',3'-d]furan moiety, while 7 and 9 were the first breviones featuring an epoxy ring and an 18-hydroxymethyl group, respectively. The anti-inflammatory and antiviral activities of compounds 1-19 were evaluated. Compounds 1, 8, 16, and 17 showed inhibitory effects against NO secretion in LPS-activated Raw264.7 macrophage cells with IC50 values ranging from 1.4 to 12.9 μM, while 1 and 7 exhibited antiviral effects against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) omicron subvariant BA.2 pseudovirus with IC50 values of 8.5 and 10.3 μM, respectively.

A New Methylbutyric Acid Derivative with Pro-angiogenesis Activity from the Mangrove Fungus Penicillium polonicum M-3.

A new methylbutyric acid derivative penianoic acid A (1), along with fifteen known compounds (2-16), was isolated and identified from the culture extract of Penicillium polonicum M-3 sourced from Malaysian mangrove root soil. Their structures were determined by detailed spectroscopic analysis of NMR and MS data and quantum chemical calculations of ECD and NMR (with DP4+ probability analysis). The isolated compounds were evaluated for pro-angiogenesis activity using zebrafish in vivo model. Compounds 1, 3, 4, 6-10, and 12-15 showed potent in pro-angiogenic activity in transgenic zebrafish.

Antiproliferative effects of hazelnut cell culture extract on the different cancer cell lines.

The increasing incidence of cancer has necessitated the discovery of novel anticancer compound sources. The presence of taxanes in hazelnut cell cultures has promoted new promising pharmacotherapeutic applications. The antiproliferative properties of hazelnut (Corylus avellana cv. 'Kalınkara') cell culture extracts against different human cancer cell lines (HeLa, MCF-7, MDA-MB-231, A549) with Beas-2B as control were evaluated. The cytotoxicity of C. avellana culture extract (5 µM, 10 µM, and 20 µM) on all cell lines was evaluated with xCELLigence Real Time Cell Analysis System. Mitotic activity (450-655 nm), BrdU activity (450-550 nm) and caspase 3,7 activity (490-520 nm) were analyzed with a spectrophotometer through 24, 48, and 72 hours. Based on the values obtained from the xCELLigence Cell Analysis System, a 10 µM concentration of the culture extract was assigned as the IC50 dose. Culture extracts at 10 µM enhanced the reduction in the proliferation of all cancer cells assayed. The highest decrease in mitotic (59.32%) and BrdU (53.77%) activity was observed in A549 lung cancer cells. However, caspase 3,7 activity (35.08%) was the highest in aggressive MDA-MB-231 breast cancer cells. The culture extracts decreased the viability of A549 cells to a greater extent than that of MCF-7 and MDA-MB-231 breast, and HeLa cervical cancer cells. C. avellana cv. 'Kalınkara' cell culture extracts have potential use in the treatment of lung and, to a lesser extent, breast and cervical cancers.

Chemical quantification of N-acyl alanine methyl ester (NAME) production and impact on temporal gene expression patterns in Roseovarius tolerans EL-164

BackgroundPrevious studies have identified structurally diverse N-acyl amino acid methyl esters (NAMEs) in culture extracts of Roseovarius tolerans EL-164 (Roseobacteraceae). NAMEs are structural analogues of the common signaling compounds N-acyl homoserine lactones (AHLs), but do not participate in AHL-mediated signaling. NAMEs show minor antialgal and antimicrobial activity, but whether this activity serves as the primary ecological role remains unclear.ResultsTo enable dose-dependent bioactivity-testing, we have established a chromatographic method for quantification of NAMEs in bacterial culture extracts. The concentrations determined for the two major NAMEs produced by EL-164, C16:1-NAME and C17:1-NAME, ranged between 0.685 and 5.731 mg L− 1 (2.0-16.9 µM) and 5.3–86.4 µg L− 1 (15.0-244.3 nM), respectively. Co-quantification of the C14:1-AHL showed concentrations ranging between 17.5 and 58.7 mg L− 1 (56.6-189.7 µM). We observed distinct production patterns for NAMEs and AHLs, with a continuous NAME production during the entire incubation period. We conducted a spike-in experiment, using the determined metabolite concentrations. By comparing the transcriptomes of pre- and post-metabolite-spikes, we identified three clusters of differentially expressed genes with distinct temporal expression patterns. Expression levels of stress response genes differed between NAME- and AHL-spiked EL-164 cultures in the stationary phase.ConclusionsOur findings support previous studies suggesting an ecological role for C16:1-NAME as antibiotic, by proving that NAME concentrations in batch cultures were higher than the minimal inhibitory concentrations against Maribacter sp. 62 − 1 (Flavobacteriia) and Skeletonema costatum CCMP 1332 (Coscinodiscophyceae) reported in the literature. Our study further exemplified the broad application range of dose-dependent testing and highlighted the different biological activities of NAMEs and AHLs.

Production of Phenyldiazene Derivatives Using the Biosynthetic Pathway of an Aromatic Diazo Group-Containing Natural Product from an Actinomycete.

The diazo group is an important functional group in organic synthesis because it confers high reactivity to the compounds and has been applied in various chemical reactions, such as the Sandmeyer reaction, Wolff rearrangement, cyclopropanation, and C-N bond formation with active methylene compounds. Previously, we revealed that 3-diazoavenalumic acid (3-DAA), which is potentially produced by several actinomycete species and contains an aromatic diazo group, is a biosynthetic intermediate of avenalumic acid. In this study, we aimed to construct a production system for phenyldiazene derivatives by adding several active methylene compounds to the culture of a 3-DAA-producing recombinant actinomycete. First, acetoacetanilide and its derivatives, which have an active methylene and are raw materials for arylide yellow dyes, were individually added to the culture of a 3-DAA-producing actinomycete. When their metabolites were analyzed, each expected compound with a phenyldiazenyl moiety was detected in the culture extract. Moreover, we established a one-pot in vitro enzymatic production system for the same phenyldiazene derivatives using a highly reactive diazotase, CmaA6. These results showed that the diazo group of natural products is an attractive tool for expanding the structural diversity of natural products both in vivo and in vitro.

In vitro, in vivo and in silico anticancer activity and toxicity of Usnic acid extracted from the mycobiont culture of Usneabaileyi.

Lichen has widespread uses in indigenous medicinal traditions, and it has been used to treat various conditions. Among the various active compounds in the lichen, Usnic acid has unique biological activities. However, the toxicology and the mechanism of action of Usnic acid need to be examined in detail to the extent of clinical trials. Hence, the present research is aimed to isolate usnic acid from the mycobiont culture of Usnea baileyi lichen and explore its toxicological properties and those of pharmacological importance. The methanolic extract of these mycobiont cultures has shown the presence of Usnic acid via column chromatography. The Usnic acid was computationally evaluated using molecular docking studies (GLIDE docking) and MMGBSA energy calculations. The results obtained from TLC, LC-MS, UV-VIS spectrophotometer, NMR spectroscopy (1H and 13c), and FT-IR indicated the presence of Usnic acid. An acute toxicity analysis by an in vivo study using a zebrafish animal model showed acute toxicity. The standard disc diffusion and DPPH assay suggested an antimicrobial and antioxidant activity with this lichen compound. The in vitro analysis of Usnic acid in G361 skin cancer cell lines demonstrated its anticancer potential against malignant melanoma. Furthermore, a molecular in silico study returned a good docking score of -7.977, and the MMGBSA free energy was measured as -18 and -30. Therefore, further extensive research on Usnic acid might lead to the development of novel anticancer drugs from the natural sources.

Efficient periplasmic expression of active lysyl endopeptidase and optimizing the purification methods

Recombinant production of lysyl endopeptidase (Lys-C) which is frequently used in proteomics is still challenging due to its complex structure. Herein, periplasmic expression and determining effective factors for recovery of the active enzyme were investigated. The codon-optimized Lys-C gene was cloned into pET26b (+) for periplasmic expression in E. coli Rosetta (DE3). The following parameters affecting expression level and activity of Lys-C were investigated including IPTG concentration (0.05–1 mM), cell density (OD600: 0.45–0.8) at induction time, presence of reducing agents (glutathione or cysteine, 0–10 mM) in culture medium or periplasmic extraction buffers, and harvesting time (6 or 20 h). Lys-C was then purified by DEAE and Ni-NTA chromatography methods. The highest expression level was obtained at 0.05 mM IPTG (5.49 %), also 8 mM cysteine, induction at OD600: 0.45 and 6 h incubation increased enzyme activity to 23.5 %, 13.3 %, and 76.4 %, respectively. The enzyme activity of Lys-C in the presence of 4 mM glutathione and extraction buffers containing 2 mM 2-mercaptoethanol (2 ME) was 81.6 % higher than the condition without reducing agents. Also, 8 mM cysteine in the culture medium and 2 mM 2 ME in extraction increased the activity up to 29.7 %. Moreover, optimization of purification process enhanced the enzyme activity from 0.217 mU to 1.76 mU. Statistical analysis showed the examined parameters significantly affected enzyme activity (p < 0.05). The presence of the reducing agents in the culture medium and extraction buffers presumably improves the Lys-C folding and increases the enzyme activity.

In vitro toxicity of microalgae species of the phyla chlorophyta and ochrophyta in CHO-k1 and HEP G2 cells for potential use in human nutrition

Microalgae are a promising component to enhance human nutrition, but to date only a few species have been authorized to be used in human nutrition. In this study, the in vitro toxicity of eight novel microalgae strains (Botryococcus braunii, Chlorococcum novae-angliae, Microchloropsis salina, Myrmecia bisecta, Stichococcus sp. Tetraselmis suecica, Tetradesmus obliquus and Spongiochloris minor) selected for their potential for human nutrition was investigated. N-hexane, acetone, ethanol and aqueous extracts of the lyophilized biomass were tested in the CHO-k1 and HEP G2 cell lines at concentrations of up to 1.81 mg ml-1 extracted biomass per well. None of the tested microalgae extracts reached values defined as a significant cytotoxic effect (IC50 &lt; 0.02 mg ml-1). The highest cytotoxic effects were measured for Stichococcus sp. in both cell lines with IC50 values of 0.17 mg ml-1 for the acetone extract in CHO-k1 cell culture and 0.18 mg ml-1 for the acetone extract in HEP G2 cell culture. Most cytotoxic effects occurred with the acetone and ethanol extracts, while the water and n-hexane extracts showed almost no measurable cytotoxic effects. Only Tetraselmis suecica showed no cytotoxic effect under the chosen conditions in both tested cell lines, marking this microalgae as particularly interesting for further investigations into its use in human nutrition.

Chemical composition of anti-microbially active fractions derived from extract of filamentous fungus Keratinophyton Lemmensii including three novel bioactive compounds

Screening for new bioactive microbial metabolites, we found a novel okaramine derivative, for which we propose the trivial name lemmokaramine, as well as two already known okaramine congeners – okaramine H and okaramine J - responsible for antimicrobial activity of the recently described microscopic filamentous fungus, Keratinophyton lemmensii BiMM-F76 (= CCF 6359). In addition, two novel substances, a new cyclohexyl denominated lemmensihexol and a new tetrahydroxypyrane denominated lemmensipyrane, were purified and characterized. The compounds were isolated from the culture extract of the fungus grown on modified yeast extract sucrose medium by means of flash chromatography followed by preparative HPLC. The chemical structures were elucidated by NMR and LC-MS. The new okaramine (lemmokaramine) exerted antimicrobial activity against Gram-positive and Gram-negative bacteria, yeasts and fungi and anticancer activity against different mammalian cell lines (Caco-2, HCT116, HT29, SW480, MCM G1, and MCM DLN). Furthermore, we found a significant antioxidant effect of lemmokaramine following H2O2 treatment indicated by activation of the Nrf2 pathway. This is the first report describing analysis and structural elucidation of bioactive metabolites for the onygenalean genus Keratinophyton.

Влияние биологически активных веществ на тепловой и окислительный стресс модельных объектов Caenorhabditis elegans

Modern medicine strives to prevent age-related diseases. Oxidative stress is associated with development and progression of various diseases. Reactive oxygen species are part of vital physiological processes. High levels of reactive oxygen lead to stress and pathology whereas low ones are associated with healthy physiology. Plant-derived adaptogens demonstrate good results in stress tolerance and homeostasis. Plant materials are a pharmacologically optimal source of chemical compounds to treat various diseases, including those caused by oxidative stress. The research featured biologically active substances isolated from extracts of callus, suspension, and root cultures of medicinal plants. Baicalin and trans-cinnamic acid were obtained from Scutellaria baicalensis while ursolic acid came from Thymus vulgaris. The biologically active substances were tested for neuroprotective properties, as well as for the impact on the expression of SOD-3 and HSP-16.2. Caenorhabditis elegans served as a model to study the accumulation of carbonylated proteins and lipofuscin. The neuroprotective activity of all tested substances decreased as their concentration fell from 200 to 10 μmol/L. C. elegans proved more resistant to thermal stress if pretreated with the biologically active substances. In response to thermal stress, nematodes expressed SOD-3 more actively than HSP-16.2. At 100 μmol/L, the biologically active substances could reduce the level of carbonylated proteins. Ursolic acid was especially effective against protein carbonylation and lipofuscin accumulation in all concentrations. Baicalin, trans-cinnamic acid, and ursolic acid made it possible to reduce oxidative and thermal stress, thus demonstrating good prospects for further studies as part of adaptogenic prepa rations.

Biological active compounds from edible mushroom: Effect on hela cells

Background: Mushrooms are unique organisms rich in proteins, carbohydrates, vitamins, and minerals, which allows them to be used not only as a source of nutrition but also in cosmetics and medicine. The fungus P. ostreatus possesses two strong enzymatic complexes – lignin-decaying and cellulose-decaying. Objective: The research aimed to study the antitumor activity of treated mm-wave mycelial extracts of mushrooms on the HeLa cell line. Research on edible mushrooms against tumors is very actual and important for the field of functional food science and bioactive compounds field. Methods: The effect of the fungal culture extract on the cyto-photometric and morphometric indices of the HeLa cell line culture was studied according to the nature of proliferation and transcription of malignant tissues. Using high-resolution liquid chromatography, the amino acid composition of acid-soluble proteins in mycelial extracts of a mushroom culture treated with mm waves was studied. Results: Extracts of P. ostreatus mycelial culture significantly suppressed the growth of proliferative activity of human tumor cultures and completely suppressed the mitotic activity of continuous HeLa cell cultures to 72 hours. Our study revealed a significant increase in glutamic and aspartic acids in mycelial culture extracts treated by mm-waves. Conclusion: Based on HPLC analysis, we suggest polyfunctional peroxidase as the candidate for the role of biologically active compound in the composition of aqueous extracts of the mycelial culture. So, the polyfunctional peroxidase plays an active role in the therapeutic properties of the oyster mushroom. Keywords: anticancer activity, ant-inflammation activity, mm-waves, mushroom`s, mycelial extracts, HPLC of acid-soluble proteins, HeLa cell line.

Investigation of Antimicrobial Compounds Produced by Endolichenic Fungi in Different Culture Media.

Continuous use of synthetic fungicides has led to explosive emergence of fungicide-resistant microbes. Therefore, there are urgent needs for environmentally friendly antimicrobial agents with novel modes of action. This study investigated endolichenic fungi (ELF) as a source of antimicrobial compounds against various plant pathogens. We utilized an One Strain MAny Compounds (OSMAC) approach to enhance the chemical diversity of fourteen ELF. This involved cultivation of ELF in four growth media and subsequently assessing antimicrobial activities of culture extracts. Nearly half of the culture extracts exhibited antimicrobial activity against a Gram-positive bacterium, but showed minimal activity against Gram-negative bacteria tested. Notably, culture extracts from two ELF, Chaetomium globosum and Nodulisporium sp., demonstrated significant inhibitory effects against plant pathogenic fungi. LC-MS/MS-based metabolome profiling confirmed the presence of known bioactive compounds like cyclic dipeptides and chaetoglobosins. These findings highlight the effectiveness of combining OSMAC and metabolomics for identifying antimicrobial agents for agricultural use.

Exploring Antimicrobial Potency, ADMET, and Optimal Drug Target of a Non-ribosomal Peptide Sevadicin from Bacillus pumilus, through In Vitro Assay and Molecular Dynamics Simulation.

The current study was designed to explore the biosynthetic potential of sevadicin in Bacillus pumilus species and its interaction with bacterial drug target molecules. The non-ribosomal peptide (NRP) cluster in B. pumilus SF-4 was preliminarily confirmed using PCR-based screening, and the bioactivity of strain SF-4 culture extract was assessed against a set of human pathogenic strains. The susceptibility assay showed that strain SF-4 extract had higher inhibitory concentrations (312-375µg/mL) than ciprofloxacin. Genome mining of B. pumilus strains (n = 22) using AntiSMASH and BAGEL identified sevadicin coding biosynthetic gene cluster only in strain SF-4, constitutes of two core biosynthetic genes, three additional biosynthetic genes, two transport-related genes, and one regulatory gene. The molecular docking of sevadicin with various putative bacterial drug targets such as dihydropteroate, muramyl ligase E, topoisomerase, penicillin-binding protein, and in vitro safety analyses were conducted with detailed ADMET screening. The results showed that sevadicin makes hydrophobic interaction with MurE (PDB ID: 1E8C and 4C13) via hydrogen bonding, suggesting bacterial growth inhibition by disrupting the cell wall synthesis pathway and exhibiting a secure biosafety profile. The stability and compactness of sevadicin/MurE complexes were assessed via molecular dynamic simulation using RMSD, RMSF, and Rg. The simulation results revealed the binding stability of sevadicin/MurE complexes and indicated that the complexes can't be easily deformed. In conclusion, the current study explored the biosynthesis of sevadicin in B. pumilus for the first time and found that sevadicin inhibits bacterial growth by inhibiting cell wall synthesis via targeting the MurE enzyme and exhibits no toxicity.

Antibacterial and Anticancer Properties of Endophenazines from Streptomyces prasinus ZO16, an Endophyte in Zingiber officinale Rosc.

&lt;b&gt;Background and Objective:&lt;/b&gt; This study investigated a bacterial strain, ZO16, isolated from ginger (&lt;i&gt;Zingiber officinale&lt;/i&gt;) roots. Analysis of its 16S ribosomal DNA (rDNA), along with chemical and physical properties, revealed it to be &lt;i&gt;Streptomyces prasinus&lt;/i&gt;. This study aimed to isolate and characterize the main bioactive compounds from ZO16, evaluating their antibacterial and anticancer properties. &lt;b&gt;Materials and Methods:&lt;/b&gt; Techniques like column chromatography and thin-layer chromatography (TLC) were used to purify the key compounds from ZO16's culture extract. Nuclear Magnetic Resonance (NMR) spectroscopy and mass spectrometry were utilized to confirm the identities of the purified compounds as endophenazine A (compound 1) and endophenazine B (compound 2). The antibacterial and anticancer properties of these compounds were then evaluated. &lt;b&gt;Results:&lt;/b&gt; The isolated compounds displayed antibacterial activity against &lt;i&gt;Staphylococcus aureus&lt;/i&gt; ATCC 25923 and Methicillin-Resistant &lt;i&gt;Staphylococcus aureus&lt;/i&gt; (MRSA). The minimum inhibitory concentration (MIC) of the isolated compounds against bacteria ranged from 8 to 32 μg/mL, while the minimum bactericidal concentration (MBC) was between 32 and 128 μg/mL. These compounds exhibited effectiveness against tested cancer cells with IC&lt;sub&gt;50&lt;/sub&gt; values ranging from 30.40 to 32.51 μg/mL for cervical cancer (HeLa), 78.32 to 86.45 μg/mL for liver cancer (HepG2) and 23.41 to 28.26 μg/mL for breast cancer (MDA-MB-231) cells. However, these compounds also showed moderate toxicity towards non-cancerous Vero cells (IC&lt;sub&gt;50&lt;/sub&gt; = 317.44-328.63 μg/mL). &lt;b&gt;Conclusion:&lt;/b&gt; The findings of this study suggest that &lt;i&gt;Streptomyces prasinus&lt;/i&gt; strain ZO16 produces compounds with antibacterial and anticancer properties. Further investigation of these compounds has the potential to contribute to the development of improved methods for controlling and treating bacterial infections and some cancers.