Intracellular Extracts Research Articles

Detoxification of Acrylamide by Potentially Probiotic Strains of Lactic Acid Bacteria and Yeast.

Some potentially probiotic strains of lactic acid bacteria (LAB) and yeast that inhabit the digestive tract of humans are known to detoxify xenobiotics, including acrylamide (AA). The objective of the subsequent research was to evaluate the AA-detoxification capability of LAB and yeast isolated from various sources. Namely, the effect of AA was tested on the growth of LAB and yeast strains, as well in the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Subsequently, the AA-binding ability of LAB and yeast was investigated in various environments, including the pH, incubation temperature, cell density, and with inanimate cells. The ability of selected LAB and yeast to reduce the genotoxicity of AA was tested on Caco-2 and Hep-G2 cell lines. The results showed that all tested strains exhibited strong resistance to AA at concentrations of 5, 10, and 50 µg/mL. Also, AA was detected in the intracellular and membrane extracts of tested strains. The most effective binding strain was Pediococcus acidilactici 16 at pH = 5, cell density = 109 CFU/mL, and incubation temperature = 37 °C (87.6% of AA removed). Additionally, all tested strains reduced the genotoxicity of AA, with the greatest reduction observed at the highest concentration of 50 µg/mL. The phenomena of detoxification by potentially probiotic strains could reduce the toxic and harmful effects of AA exposure to humans every day.

Neuroprotective Effects Exerted by a Combination of Selected Lactic Acid Bacteria in a Mouse Parkinsonism Model under Levodopa-Benserazide Treatment.

Alterations of the microbiota-gut-brain axis has been associated with intestinal and neuronal inflammation in Parkinson's disease (PD). The aim of this work was to study some mechanisms associated with the neuroprotective effect of a combination (MIX) of lactic acid bacteria (LAB) composed by Lactiplantibacillus plantarum CRL2130 (riboflavin overproducing strain), Streptococcus thermophilus CRL808 (folate producer strain), and CRL807 (immunomodulatory strain) in cell cultures and in a chronic model of parkinsonism induced with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in aged mice, and under levodopa-benserazide treatment. In vitro, N2a differentiated neurons were exposed to the neurotoxin 1-methyl-4-phenylpyridinium (MPP+) and treated with intracellular bacterial extracts or with conditioned media from BV-2 cells exposed to the bacterial extracts. In vivo, motor skills, tyrosine hydrolase (TH) in brain and cytokine concentrations in serum and in brain were evaluated. The study of the faecal microbiota and the histology of the small intestine was also performed. The results showed that the neuroprotective effect associated with LAB MIX administration did not interfere with levodopa-benserazide treatment. This effect could be associated with the antioxidant and immunomodulatory potential of the LAB selected in the MIX, and was associated with the significant improvement in the motor tests and a higher number of TH + cells in the brain. In addition, LAB MIX administration was associated with modulation of the immune response. LAB administration decreased intestinal damage with an increase in the villus length /crypt depth ratio. Finally, the administration of the LAB MIX in combination with levodopa-benserazide treatment was able to partially revert the intestinal dysbiosis observed in the model, showing greater similarity to the profiles of healthy controls, and highlighting the increase in the Lactobacillaceae family. Different mechanisms of action would be related to the protective effect of the selected LAB combination which has the potential to be evaluated as an adjuvant for conventional PD therapies.

Study on Optimization of Liquid Fermentation Medium and Antitumor Activity of the Mycelium on Phyllopora lonicerae.

Phylloporia lonicerae is an annual fungus that specifically parasitizes living Lonicera plants, offering significant potential for developing new resource food and medicine. However, wild resources and mycelium production of this fungus is limited, and its anti-tumor active ingredients and mechanisms remain unclear, hampering the development of this fungus. Thus, we optimized the fermentation medium of P. lonicerae and studied the anti-tumor activity of its mycelium. The results indicated that the optimum fermentation medium consisted of 2% sucrose, 0.2% peptone, 0.1% KH2PO4, 0.05% MgSO4·7H2O, 0.16% Lonicera japonica petals, 0.18% P fungal elicitor, and 0.21% L. japonica stem. The biomass reached 7.82 ± 0.41 g/l after 15 days of cultivation in the optimized medium, a 142% increase compared with the potato dextrose broth medium, with a 64% reduction in cultivation time. The intracellular alcohol extract had a higher inhibitory effect on A549 and Eca-109 cells than the intracellular water extract, with half-maximal inhibitory concentration values of 2.42 and 2.92 mg/ml, respectively. Graded extraction of the alcohol extract yielded petroleum ether phase, chloroform phase, ethyl acetate phase, and n-butanol phase. Among them, the petroleum ether phase exhibited a better effect than the positive control, with a half-maximal inhibitory concentration of 113.3 μg/ml. Flow cytometry analysis indicated that petroleum ether components could induce apoptosis of Eca-109 cells, suggesting that this extracted component can be utilized as an anticancer agent in functional foods. This study offers valuable technical support and a theoretical foundation for promoting the comprehensive development and efficient utilization of P. lonicerae.

In vitro antioxidant and anticancer potential of intra-cellular ethyl acetate extract of marine-derived fungus Talaromyces tratensis SS10

Marine fungi are well-known for producing structurally distinct secondary metabolites, making them potential sources of novel therapies. The present investigation aims to study the in vitro antioxidant and anticancer potential of intra-cellular crude ethyl acetate extracts of Talaromyces tratensis SS10. In the present study, qualitative and quantitative phytochemical studies of various solvent extracts of T. tratensis have been carried out using standard protocols. Later, ethyl acetate extract of T. tratensis was analyzed for phytochemicals using Gas Chromatography Mass Spectrometry (GC-MS). Further, the antioxidant properties of the T. tratensis ethyl acetate extract have been done by Ferric reducing antioxidant power assay (FRAP). Further, the anticancer potential of this extract has been carried out by MTT assay against human cancer cells such as MDA MB 231, HeLa, and HT-29. Ethyl acetate exhibited a higher yield of chemical extraction than the other solvents used. The GCMS analysis of T. tratensis ethyl acetate extract revealed major phytoconstituents such as N-(1,1-Dimethylpropyl)-2,2,3-trimethylaziridine-1-carboxamide, 1-Undecanol, 5,5 Dimethyl-3-vinyl cyclohex-2-en-1-one, 1,2-Benzenedicarboxylic acid, bis (2-methyl propyl) ester. T. tratensis ethyl acetate extract showed the highest percentage of Fe3+ reduction (48.093±1.469%) at 120 μg/mL, with an IC50 value of 157.26 μg/mL concentration. Furthermore, 100 μg/mL of the extract showed significant cell death rates in cytotoxic assays, indicating a low percentage of viable cells for all three examined cell lines. The T. tratensis ethyl acetate extract has shown a dose-dependent cytotoxic effect against all tested cancer cell lines. The better IC50 value (6.25 μg/mL) was recorded in the case of HeLa cell lines followed by 12.5 μg/mL for both MDA MB 231 and HT-29 cell lines. The presence of bioactive compounds such as Benzeneethanamine, N-[(pentafluorophenyl)methylene]-beta.,3,4-tris[(trimethylsilyl) oxy]-, 1,2-Benzenedicarboxylic acid, bis(2-methylpropyl) ester, and cyclononasiloxane, octadecamethyl- may have contributed to the ethyl acetate extracts’ strong antioxidant and anticancer properties. The current study’s findings show that T. tratensis SS10 has the potential for drug development due to its chemical constituents, which possess various biological activities.

Evaluating The Cytotoxic Activity of Lactobacillus plantarum IIA-1A5 Against MCF-7 Human Breast Cancer Cells and Identifying Its Surface Layer Protein Gene

Breast cancer is a serious global health concern, with a high mortality rate worldwide. Using natural compounds as potential cancer therapies is a promising approach to address this issue. Previous research has shown that probiotic lactic acid bacteria (LAB) and their metabolites, such as surface layer protein (slp), have a positive impact on a variety of health disorders, including cancer. The purpose of this study was to evaluate the ability of Lactobacillus plantarum IIA-1A5 to suppress the growth of the MCF-7 breast cancer cell line and detect the presence of the slp gene. Intracellular and extracellular protein fractions were isolated from L. plantarum IIA-1A5 cultures. The protein concentrations and molecular weights of the extracts were measured. The anticancer activity of the extracts was assessed using the MTT cytotoxicity test, and IC50 values were calculated. The slp gene was identified through polymerase chain reaction (PCR) amplification and nucleotide sequencing. The results demonstrated that L. plantarum IIA-1A5 had a concentration-dependent inhibitory effect on MCF-7 breast cancer cells, with IC50 values of 6.831 and 12.35 μg/mL for intracellular extracts and extracellular extracts, respectively. Additionally, PCR amplification and nucleotide sequencing confirmed the presence of the slp gene, which may contribute to the strain’s anticancer abilities. These findings suggest the potential of L. plantarum IIA-1A5 as a natural anticancer agent against MCF-7 breast cancer cells. Further research is warranted to elucidate the underlying mechanisms of L. plantarum IIA-1A5 in breast cancer treatment.

Enhanced potent immunosuppression of intracellular adipose tissue-derived stem cell extract by priming with three-dimensional spheroid formation

Immunomodulatory properties of mesenchymal stem cells are widely studied, supporting the use of MSCs as cell-based therapy in immunological diseases. This study aims to generate cell-free MSC extract and improves their immunomodulatory potential. Intracellular extracts were prepared from adipose-derived stem cells (ADSC) spheroid via a freeze-thawing method. The immunomodulatory capacities of ADSC spheroid extracts were investigated in vitro, including lymphocyte proliferation, T regulatory cell expansion, and macrophage assays. A comparative study was conducted with ADSC monolayer extract. The key immunomodulatory mediators presented in ADSC extract were identified. The results revealed that ADSC spheroid extract could suppress lymphocyte activation while enhancing T regulatory cell expansion. Immunomodulatory molecules such as COX-2, TSG-6, and TGF-β1 were upregulated in ADSC priming via spheroid culture. Selective inhibition of COX-2 abrogates the effect of ADSC extract on inducing T regulatory cell expansion. Thus, ADSC spheroid extract gains high efficacy in regulating the immune responses which are associated in part by COX-2 generation. Furthermore, ADSC spheroid extract possessed a potent anti-inflammation by manipulation of TNF-α production from LPS-activated macrophage. Our current study has highlighted the opportunity of using cell-free extracts from adipose tissue-derived mesenchymal stem cells spheroid as novel immunomodulators for the treatment of immunological-associated diseases.

Purification, characterisation and immobilisation of a protease‐resistant α‐galactosidase from Monascus purpureus and its application in removal of raffinose family oligosaccharides

SummaryIn this study, an α‐galactosidase (MPG) was produced by submerged fermentation from Monascus purpureus, a food‐grade fungus used for making red yeast rice. MPG was purified from intracellular extract which showed a subunit molecular mass of 95 kDa. MPG was optimally active at pH 4.0 and 50 °C and was stable between pH 3.5–8.0 and at temperatures up to 50 °C. The enzyme retained its activity in the presence of most of the metal ions except Cu2+ and Fe2+. MPG exhibited remarkable resistance to the proteases, pepsin, trypsin and proteinase K as well as to the sugars, galactose and sucrose. Moreover, it retained complete activity under simulated gastric conditions. The enzyme displayed specificity for the substrates, melibiose, raffinose and stachyose and was able to hydrolyze these oligosaccharides. It was effective in the removal of non‐digestible raffinose family oligosaccharides (RFOs) from soybeans, chickpeas and kidney beans. MPG was immobilised onto chitosan and the immobilised enzyme (iMPG) displayed higher optimal temperature (60 °C), better thermostability and substrate specificity as compared to MPG. iMPG was also capable of hydrolyzing RFOs in chickpeas and could be efficiently reused six times. In view of their superior properties, MPG and iMPG may have great prospects in the food and feed industries for the removal of RFOs from soy and other legumes.

The antioxidant activity and metabolomic analysis of the supernatant of Streptococcus alactolyticus strain FGM

Strain-specific probiotics can present antioxidant activity and reduce damage caused by oxidation. Streptococcus alactolyticus strain FGM (S. alactolyticus strain FGM) isolated from the chicken cecum shows potential probiotic properties which have been previously demonstrated. However, the antioxidant properties of S. alactolyticus strain FGM remain unknown. In this view, cell-free supernatant (CFS), intact cells (IC) and intracellular extracts (CFE) of strain FGM and 3 strains of Lactobacillus (LAB) were prepared, and their scavenging capacities against DPPH, hydroxyl radicals and linoleic acid peroxidation inhibitory were compared in this study. The effects of strain FGM cell-free supernatant (FCFS) on NO production, activity of SOD and GSH-Px in RAW264.7 cells and LPS-induced RAW264.7 cells were analyzed. The metabolites in the supernatant were quantitated by N300 Quantitative Metabolome. It was shown that the physicochemical characteristics of CFS to scavenge DPPH, hydroxyl radicals, and linoleic acid peroxidation inhibitory were significantly stronger than that of IC and CFE in the strain FGM (P < 0.05), respectively 87.12% ± 1.62, 45.03% ± 1.27, 15.63% ± 1.34. FCFS had a promotional effect on RAW264.7 cells, and significantly elevated SOD and GSH-Px activities in RAW264.7 cells. 25 μL FCFS significantly promoted the proliferation of RAW264.7 cells induced by LPS, increased the activities of SOD and GSH-PX, and decreased the release of NO. Furthermore, among the differential metabolites of FCFS quantified by N300, 12 metabolites were significantly up-regulated, including lactic acid, indole lactic acid, linoleic acid, pyruvic acid etc., many of which are known with antioxidant properties. In conclusion, FCFS had good antioxidant properties and activity, which can be attributed to metabolites produced from strain FGM fermentation. It was further confirmed that S. alactolyticus strain FGM and its postbiotic have potential probiotic properties and bright application prospects in livestock and poultry breeding.

Identification of a Novel Peptide with Alcohol Dehydrogenase Activating Ability from Ethanol-Induced Lactococcus lactis: A Combined In Silico Prediction and In Vivo Validation.

Alcohol dehydrogenase (ADH) is a crucial rate-limiting enzyme in alcohol metabolism. Our previous research found that ethanol-induced intracellular extracts of Lactococcus lactis (L. lactis) could enhance alcohol metabolism in mice, but the responsible compounds remain unidentified. The study aimed to screen potential ADH-activating peptides from ethanol-induced L. lactis using virtual screening and molecular docking calculation. Among them, the pentapeptide FAPEG might bind to ADH through hydrophobic interaction and hydrogen bonds, then enhancing ADH activity. Spectroscopy analysis further investigated the peptide-enzyme interaction between FAPEG and ADH, including changes in the amino acid residue microenvironment and secondary structural alterations. Furthermore, FAPEG could protect against alcoholic liver injury (ALI) in mice by reducing blood alcohol concentration, enhancing the activity of antioxidant and alcohol metabolism enzymes, and attenuating alcohol-induced hepatotoxicity, which was related to the activation of the Nrf2/keap1/HO-1 signaling pathway. The study provided preliminary evidence that the generation of ADH-activating peptides in ethanol-induced L. lactis has the potential in preventing ALI in mice using in silico prediction and in vivo validation approaches.

Water-soluble intracellular extract of Desmodesmus sp. YT enhanced the antioxidant capacity of human skin fibroblast to protect the skin from UV damage.

The oxidative stress induced by ultraviolet (UV) radiation is a pivotal factor in skin aging and can even contribute to the development of skin cancer. This study explored the antioxidant effect and mechanism of water-soluble intracellular extract (WIE) of Desmodesmus sp.YT (YT), aiming to develop a natural antioxidant suitable for incorporation into cosmetics. The study evaluated the scavenging capacity of YT-WIE against free radicals and assessed its impact on human skin fibroblasts (HSF) cell viability and UV resistance using Cell Counting Kit-8 (CCK-8). Transcriptome sequencing was employed to elucidate the mechanism of action, while RT-qPCR and western blot were used to validate the expression of key genes. YT-WIE displayed robust antioxidant activity, demonstrating potent scavenging abilities against 2,2-diphenyl-1-picrylhydrazyl (DPPH; IC50 = 0.55 mg mL-1), 2,2'-Azino-bis (3 ethylbenzothiazoline-6-sulfonic acid; ABTS; IC50 = 3.11 mg mL-1), Hydroxyl (·OH; IC50 = 2.21 mg mL-1), and Superoxide anion (O2 •-; IC50 = 0.98 mg mL-1). Furthermore, compared to the control group, the YT-WIE group exhibited an 89.30% enhancement in HSF viability and a 44.63% increase in survival rate post-UV irradiation. Significant upregulation of antioxidant genes (GCLC, GCLM, TXNRD1, HMOX1, NQO1) was observed with YT-WIE treatment at 400 μg mL-1, with fold increases ranging from 1.13 to 5.85 times. YT-WIE demonstrated considerable potential as an antioxidant, shielding human cells from undue oxidative stress triggered by external stimuli such as UV radiation. This suggests its promising application in cosmetics antioxidants.

Semi-industrial Bio-fabrication of ZnO/MnO2 Nanocomposite Using Endophytic Streptomyces coelicolor: Characterization, Statistical Design, Exponential Pulse Fed-Batch Fermentation, and Its Antimicrobial Application

In our study, we examined how well six Streptomyces strains bio-fabricated ZnONPs, MnONPs, and/or ZnO/MnO2 nanocomposite. The most potent strain that generated efficient antimicrobial nanoparticles was then picked to increase the production of those particles in a semi-industrial pilot plant unit. Consequently, the intracellular extract of endophytic Streptomyces coelicolor strain E72 was used to achieve the bio-fabrication reaction of the spherical ZnO/MnO2 nanocomposite (6–18 nm). The bio-fabricated ZnO/MnO2 nanocomposite was validated and characterized using FTIR, XRD, SEM, TEM, TGA, and EDS analyses. Additionally, the production of this ZnO/MnO2 nanocomponent was scaled up to a pilot plant unit with a semi-industrial size. The Plackett–Burman experimental method was used to maximize the production of ZnO/MnO2 nanocomposites, which had increased 2.7-fold from their initial state. The bio-fabricated ZnO/MnO2 nanocomposite was subsequently scaled up 31.25 times using an exponential pulse-feeding fermentation technique in a 70-L bioreactor. This ZnO/MnO2 nanocomposite exhibited effective antimicrobial efficacy against all tested antibiotic-resistant human pathogens. The antimicrobial effects against Salmonella paratyphi (53.17 ± 2.8 mm) and Candida albicans (50.2 ± 1.01 mm) were the most potent at 90 and 130 µg/ml of ZnO/MnO2 nanocomposite, respectively. This is the first full explanation of the ZnO/MnO2 nanocomposite bio-fabrication at a semi-industrial scale employing endophytic strain E72 extract as a reducing/capping agent that reacted with MnCl2·4H2O and Zn (CH3COO)2·2H2O as precursors. This bio-fabricated ZnO/MnO2 nanocomposite has the potential to be utilized in the development of pharmaceuticals, cosmetics, wound dressings, and burn therapy due to its powerful antimicrobial capabilities.

Studying the O-GlcNAcome of human placentas using banked tissue samples.

O-GlcNAcylation is a dynamic modulator of signaling pathways, equal in magnitude to the widely studied phosphorylation. With the rapid development of tools for its detection at the single protein level, the O-GlcNAc modification rapidly emerged as a novel diagnostic and therapeutic target in human diseases. Yet, mapping the human O-GlcNAcome in various tissues is essential for generating relevant biomarkers. In this study, we used human banked tissue as a sample source to identify O-GlcNAcylated protein targets relevant to human diseases. Using human term placentas, we propose (1) a method to clean frozen banked tissue of blood proteins; (2) an optimized protocol for the enrichment of O-GlcNAcylated proteins using immunoaffinity purification; and (3) a bioinformatic workflow to identify the most promising O-GlcNAc targets. As a proof-of-concept, we used 45mg of banked placental samples from two pregnancies to generate intracellular protein extracts depleted of blood protein. Then, antibody-based O-GlcNAc enrichment on denatured samples yielded over 2000 unique HexNAc PSMs and 900 unique sites using 300μg of protein lysate. Due to efficient sample cleanup, we also captured 82 HexNAc proteins with high placental expression. Finally, we provide a bioinformatic tool (CytOVS) to sort the HexNAc proteins based on their cellular localization and extract the most promising O-GlcNAc targets to explore further. To conclude, we provide a simple 3-step workflow to generate a manageable list of O-GlcNAc proteins from human tissue and improve our understanding of O-GlcNAcylation's role in health and diseases.

Deciphering the human antibody response against Burkholderia pseudomallei during melioidosis using a comprehensive immunoproteome approach.

The environmental bacterium Burkholderia pseudomallei causes the often fatal and massively underreported infectious disease melioidosis. Antigens inducing protective immunity in experimental models have recently been identified and serodiagnostic tools have been improved. However, further elucidation of the antigenic repertoire of B. pseudomallei during human infection for diagnostic and vaccine purposes is required. The adaptation of B. pseudomallei to very different habitats is reflected by a huge genome and a selective transcriptional response to a variety of conditions. We, therefore, hypothesized that exposure of B. pseudomallei to culture conditions mimicking habitats encountered in the human host might unravel novel antigens that are recognized by melioidosis patients. In this study, B. pseudomallei was exposed to various stress and growth conditions, including anaerobiosis, acid stress, oxidative stress, iron starvation and osmotic stress. Immunogenic proteins were identified by probing two-dimensional Western blots of B. pseudomallei intracellular and extracellular protein extracts with sera from melioidosis patients and controls and subsequent MALDI-TOF MS. Among B. pseudomallei specific immunogenic signals, 90 % (55/61) of extracellular immunogenic proteins were identified by acid, osmotic or oxidative stress. A total of 84 % (44/52) of intracellular antigens originated from the stationary growth phase, acidic, oxidative and anaerobic conditions. The majority of the extracellular and intracellular protein antigens were identified in only one of the various stress conditions. Sixty-three immunoreactive proteins and an additional 38 candidates from a literature screening were heterologously expressed and subjected to dot blot analysis using melioidosis sera and controls. Our experiments confirmed melioidosis-specific signals in 58 of our immunoproteome candidates. These include 15 antigens with average signal ratios (melioidosis:controls) greater than 10 and another 26 with average ratios greater than 5, including new promising serodiagnostic candidates with a very high signal-to-noise ratio. Our study shows that a comprehensive B. pseudomallei immunoproteomics approach, using conditions which are likely to be encountered during infection, can identify novel antibody targets previously unrecognized in human melioidosis.

Assessment of mutagenic activity of phlorotannin-enriched extracts of three brown algal species

BACKGROUND: Phlorotannins are unique phenolic compounds produced by brown algae. Due to their considerable biological activity these metabolites are extensively studied in the context of medicinal applications. However, to date, no studies addressed potential genotoxicity of phlorotannins.&#x0D; AIM: The objective of this research is an assessment of mutagenic activity of intracellular and cell wall (CW) bound phlorotannins of three brown algal species.&#x0D; MATERIALS AND METHODS: Mutagenicity of phlorotannin extracts of Desmarestia aculeata, Fucus serratus, and Ectocarpus siliculosus was assessed by the Ames test, carried out using three tester strains of Salmonella typhimurium (TA97, TA98, and TA100) with and without metabolic activation.&#x0D; RESULTS: Intracellular phlorotannin extracts of all tested algae showed relatively low values of minimum inhibitory concentration against S. typhimurium (20–30 μg/ml), with extract of D. aculeata being the most toxic. Intracellular phlorotannins of F. serratus and CW-bound polyphenols of E. siliculosus demonstrated moderate mutagenic activity in the Ames test inducing frameshift mutations with the number of His+ revertants more than twice higher compared to the control. The phlorotannin extracts of D. aculeata showed no mutagenic activity.&#x0D; CONCLUSIONS: The brown alga D. aculeata may be regarded as a promising source of phlorotannins for medical applications, as its phlorotannin-enriched extracts have high antibiotic activity and are not mutagenic.

Optimization of the biosynthesis of silver nanoparticles using bacterial extracts and their antimicrobial potential

In the present study, silver nanoparticles (AgNPs) were biosynthesized using the supernatant and the intracellular extract of Cupriavidus necator, Bacillus megaterium, and Bacillus subtilis. The characterization of the AgNPs was carried out using UV–Vis spectroscopy, FTIR, DLS and TEM. Resazurin microtiter-plate assay was used to determine the antimicrobial action of AgNPs against Escherichia coli. UV–Visible spectra showed peaks between 414 and 460 nm. TEM analysis revealed that the synthesized AgNPs showed mostly spherical shapes. DLS results determined sizes from 20.8 to 118.4 nm. The highest antimicrobial activity was obtained with the AgNPs synthesized with supernatant rather than those using the intracellular extract. Therefore, it was determined that the bacterial species, temperature, pH, and type of extract (supernatant or intracellular) influence the biosynthesis. This synthesis thus offers a simple, environmentally friendly, and low-cost method for the production of AgNPs, which can be used as antibacterial agents.

Valorization of Grape Pomace for Trametes versicolor Mycelial Mass and Polysaccharides Production

Polysaccharides and protein–polysaccharide complexes produced from the fungal strain Trametes versicolor have demonstrated bioactive properties that depend on the substrate, the fermentation conditions and also the fungal strain. Likewise, the submerged and controlled fermentation of medicinal mushrooms elicits numerous advantages over traditional processes to produce mycelia and added-value products, along with the exploitation of biodiversity. This study evaluated the growth profile of an indigenous T. versicolor isolate using commercial nutrients that were subsequently replaced with renewable resources, specifically, grape pomace extract (GPE), under static and shaking conditions. The effect of elicitor addition was also assessed using GPE. The process productivity was significantly improved, yielding 21 g/L of biomass. Agitation proved beneficial for all examined cases regarding biomass productivity and substrate consumption rates. The chemical and antioxidant profile of crude intracellular and extracellular polysaccharides was determined, whereby intracellular extracts indicated &gt;50% antioxidant activity. FTIR analysis validated the preliminary chemical characterization of the extracts, whereas the amino acid profile of IPS extracts was also included. Evidently, our study elaborates on the development of a bioconversion concept to valorize wine-making side-streams to formulate added-value products with potential bioactive attributes.

Cr(VI) reduction by Agrobacterium sp. Cr-1 and Lysinibacillus sp. Cr-2, novel Cr(VI)-reducing strains isolated from chromium plant soil.

The bioremediation of Cr(VI)-contaminated soil is a promising strategy; however, the performance of Cr(VI)-reducing bacteria is limited by the toxicity of Cr(VI). In this study, two novel Cr(VI)-reducing bacteria were isolated from a Cr salt plant and identified as Agrobacterium sp. and Lysinibacillus sp. The Cr(VI) reduction conditions of the two strains were optimized. At a Cr(VI) concentration of 500mg/L, Agrobacterium sp. Cr-1 reduced Cr(VI) with a removal rate of 96.91%, while that for Lysinibacillus sp. Cr-2 was 92.82%. First-order reaction kinetic equations simulated the positive relationship between time and Cr(VI) concentration during Cr(VI) reduction in these two strains. Agrobacterium sp. Cr-1 was further studied, and the effects of different cell components on Cr(VI) reduction were detected. The extracellular extracts of Agrobacterium sp. Cr-1 played a major role in Cr(VI) reduction, followed by intracellular extracts and cell membranes. The scanning electron microscope-energy dispersive spectrometer (SEM-EDS) images show that the precipitation was Cr. The high Cr(VI) reducing ability of Agrobacterium sp. Cr-1 suggests that this strain is promising for the remediation of Cr(VI)-contaminated sites.

Assessment and identification of bioactive metabolites from terrestrial Lyngbya spp. responsible for antioxidant, antifungal, and anticancer activities.

Lyngbya from fresh and marine water produces an array of pharmaceutically bioactive therapeutic compounds. However, Lyngbya from agricultural soil is still poorly investigated. Hence, in this study, the bioactive potential of different Lyngbya spp. extract was explored. Intracellular petroleum ether extract of L. hieronymusii K81 showed the highest phenolic content (626.22 ± 0.65 μg GAEs g-1 FW), while intracellular ethyl acetate extract of L. aestuarii K97 (74.02 ± 0.002 mg QEs g-1 FW) showed highest flavonoid content. Highest free radical scavenging activity in terms of ABTS•+ was recorded in intracellular methanolic extract of Lyngbya sp. K5 (97.85 ± 0.068%), followed by L. wollei K80 (97.22 ± 0.059%) while highest DPPH• radical scavenging activity observed by intracellular acetone extract of Lyngbya sp. K5 (54.59 ± 0.165%). All the extracts also showed variable degrees of antifungal activities against Fusarium udum, F. oxysporum ciceris, Colletotrichum capsici, and Rhizoctonia solani. Further, extract of L. wollei K80 and L. aestuarii K97 showed potential anticancer activities against MCF7 (breast cancer) cell lines. GC-MS analyses of intracellular methanolic extract of L. wollei K80 showed the dominance of PUFAs with 9,12,15-octadecatrienoic acid, methyl ester, (Z,Z,Z) as the most abundant bioactive compound. On the other hand, the extracellular ethyl acetate extract of L. aestuarii K97 was rich in alkanes and alkenes with 1-hexyl-2-nitrocyclohexane as the most predominant compound. Extracts of Lyngbya spp. rich in novel secondary metabolites such as PUFAs, alkanes, and alkenes can be further explored as an alternative and low-cost antioxidant and potential apoptogens for cancer therapy.

Promising bioactive metabolites of mangrove inhabitant Streptomyces tauricus and prostate cancer PC3 cell inhibition by antimicrobial peptides.

Streptomyces is a group of microbes known for antibiotic production and has contributed to more than 70% of present commercially available antibiotics. These antibiotics are important in the management, protection, and treatment of chronic illnesses. In the present study, the isolated S. tauricus strain from mangrove soil in Mangalore, India (GenBank accession number: MW785875) was subjected for differential cultural characterization, phenotype involving brown pigmentation, filamentous mycelia, and ash-colored spore production was observed using field emission scanning electron microscopy (FESEM) analysis revealing filamentous mycelia possessing a straight spore chain. Spores were visualized as elongated, rod-shaped, smooth surfaces with curved edges. After optimized growth conditions for S. tauricus on starch-casein agar medium, the GC/MS analysis of S. tauricus intracellular extract detected bioactive compounds reported for pharmacological applications. Analyzed using the NIST library, most of the bioactive compounds identified in intracellular extract had molecular weights of less than 1 kDa. On the PC3 cell line, the Sephadex G-10 partially purified eluted peak protein fraction demonstrated significant anticancer activity. The LCMS analysis revealed the presence of Tryprostatin B, Fumonisin B1, Microcystin LR, and Surfactin C with molecular weights below 1 kDa. This study found that small molecular weight microbial compounds are more effective in a variety of biological applications.

Whole Genome Sequence of Lactiplantibacillus plantarum HOM3204 and Its Antioxidant Effect on D-Galactose-Induced Aging in Mice.

Lactiplantibacillus plantarum, previously named Lactobacillus plantarum, is a facultative, homofermentative lactic acid bacterium widely distributed in nature. Several Lpb. plantarum strains have been demonstrated to possess good probiotic properties, and Lpb. plantarum HOM3204 is a potential probiotic strain isolated from homemade pickled cabbage plants. In this study, whole-genome sequencing was performed to acquire genetic information and predict the function of HOM3204, which has a circular chromosome of 3,232,697 bp and two plasmids of 48,573 and 17,060 bp, respectively. Moreover, various oxidative stress-related genes were identified in the strain, and its antioxidant activity was evaluated in vitro and in vivo. Compared to reference strains, the intracellular cell-free extracts of Lpb. plantarum HOM3204 at a dose of 1010 colony-forming units (CFU)/ml in vitro exhibited stronger antioxidant properties, such as total antioxidant activity, 2,2-diphenyl-1-picrylhydrazyl radical scavenging rate, superoxide dismutase activity, and glutathione (GSH) content. Daily administration of 109 CFU Lpb. plantarum HOM3204 for 45 days significantly improved the antioxidant function by increasing the glutathione peroxidase activity in the whole blood and GSH concentration in the livers of D-galactose-induced aging mice. These results suggest that Lpb. plantarum HOM3204 can potentially be used as a food ingredient with good antioxidant properties.