Bioactive Naphthoquinone and Phenazine Analogs from the Endophytic Streptomyces sp. PH9030 as α-Glucosidase Inhibitors.

A talented endophytic Streptomyces sp. PH9030 is derived from the medicinal plant Kadsura coccinea (Lem.) A.C. Smith. The undescribed naphthoquinone naphthgeranine G (5) and seven previously identified compounds, 6-12, were obtained from Streptomyces sp. PH9030. The structure of 5 was identified by comprehensive examination of its HRESIMS, 1D NMR, 2D NMR and ECD data. The inhibitory activities of all the compounds toward α-glucosidase and their antibacterial properties were investigated. The α-glucosidase inhibitory activities of 5, 6, 7 and 9 were reported for the first time, with IC50 values ranging from 66.4 ± 6.7 to 185.9 ± 0.2 µM, as compared with acarbose (IC50 = 671.5 ± 0.2 µM). The molecular docking and molecular dynamics analysis of 5 with α-glucosidase further indicated that it may have a good binding ability with α-glucosidase. Both 9 and 12 exhibited moderate antibacterial activity against methicillin-resistant Staphylococcus aureus, with minimum inhibitory concentration (MIC) values of 16 µg/mL. These results indicate that 5, together with the naphthoquinone scaffold, has the potential to be further developed as a possible inhibitor of α-glucosidase.

Dermacozine N, the First Natural Linear Pentacyclic Oxazinophenazine with UV-Vis Absorption Maxima in the Near Infrared Region, along with Dermacozines O and P Isolated from the Mariana Trench Sediment Strain Dermacoccus abyssi MT 1.1T.

Three dermacozines, dermacozines N-P (1-3), were isolated from the piezotolerant Actinomycete strain Dermacoccus abyssi MT 1.1T, which was isolated from a Mariana Trench sediment in 2006. Herein, we report the elucidation of their structures using a combination of 1D/2D NMR, LC-HRESI-MSn, UV-Visible, and IR spectroscopy. Further confirmation of the structures was achieved through the analysis of data from density functional theory (DFT)-UV-Visible spectral calculations and statistical analysis such as two tailed t-test, linear regression-, and multiple linear regression analysis applied to either solely experimental or to experimental and calculated 13C-NMR chemical shift data. Dermacozine N (1) bears a novel linear pentacyclic phenoxazine framework that has never been reported as a natural product. Dermacozine O (2) is a constitutional isomer of the known dermacozine F while dermacozine P (3) is 8-benzoyl-6-carbamoylphenazine-1-carboxylic acid. Dermacozine N (1) is unique among phenoxazines due to its near infrared (NIR) absorption maxima, which would make this compound an excellent candidate for research in biosensing chemistry, photodynamic therapy (PDT), opto-electronic applications, and metabolic mapping at the cellular level. Furthermore, dermacozine N (1) possesses weak cytotoxic activity against melanoma (A2058) and hepatocellular carcinoma cells (HepG2) with IC50 values of 51 and 38 µM, respectively.

1-hydroxy-7-oxolavanducyanin and Δ7″,8″-6″-hydroxynaphthomevalin from Streptomyces sp. CPCC 203577.

Lavanducyanin is a bioactive phenazine-containing secondary metabolite, and naphthomevalin is an antibacterial polyketide secondary metabolite. Herein, new analogues of lavanducyanin (2) and of naphthomevalin (4), together with lavanducyanin (1) and naphthomevalin (3), were identified from Streptomyces sp. CPCC 203577, an actinomycete soil isolate. The structures of 2 and 4 were elucidated as 1-hydroxy-7-oxolavanducyanin and Δ7″,8″-6″-hydroxynaphthomevalin, respectively, by 1D and 2D NMR. Antibacterial assays revealed that 2 had significant but reduced anti-Gram-positive bacterial activity compared with 1, and 4 was devoid of anti-Gram-positive bacterial activity. This indicated that the phenazinone nucleus in lavanducyanin and the monoterpene side chain in naphthomevalin might be important for their anti-Gram-positive bacterial activity. Compounds 1-4 were all inactive against Gram-negative bacteria.

Isolation and Characterization of New Phenazine Metabolites with Antifungal Activity against Root-Rot Pathogens of Panax notoginseng from Streptomyces.

Three new phenazine metabolites, strepphenazine A-C (1-3), along with a known compound baraphenazine E 4 were isolated from the culture broth of a Streptomyces strain YIM PH20095. The structures were elucidated based on the spectral data. Compounds 1-4 showed different antifungal activity against Fusarium oxysporum, Plectosphaerella cucumerina, Alternaria panax, and Phoma herbarum, which caused root-rot disease of Panax notoginseng with minimal inhibitory concentrations (MICs) of 16-64 µg/mL; compared with compound 4, compounds 1-3 showed better antifungal activity against some of these pathogenic fungi with MICs of 16-32 µg/mL, while compound 4 showed antifungal activity against F. oxysporum, P. cucumerina, and A. panax with the same MICs of 64 µg/mL. Thus, strain YIM PH20095 provides new sources for the development of biological control agents to prevent the infection of pathogenic fungi of P. notoginseng.

Basic red 2 and methyl violet adsorption by date pits: adsorbent characterization, optimization by RSM and CCD, equilibrium and kinetic studies.

The potential of raw date pits as a natural, widely available and low-cost agricultural waste has been studied in order to adsorb cationic dyes from an aqueous solution. Date pits were characterized by FTIR, SEM, BET, and XRD analysis. To optimize removal of two industrial dyes, basic red 2 (BR2) and methyl violet (MV), from aqueous solution using date pits, response surface methodology (RSM) is employed. Tests were carried out as per central composite design (CCD) with four input parameters namely contact time, temperature, initial concentration of adsorbate, and pH. Second-order polynomial model better fits experimental data for BR2 and MV and optimum values were then determined. In the optimum conditions, kinetic study was conducted and the pseudo-second-order model was found the best fitted model compared to pseudo-first-order model. Moreover, it was shown that intraparticle diffusion was not the sole controlling step and could be associated with other transfer resistance. On other hand, equilibrium isotherms were obtained for BR2 and MV and their maximum adsorption capacities were 92 and 136 mg g-1 respectively. Two-parameter isotherm models like Langmuir, Temkin, Freundlich, Dubinin-Radushkevich, and Halsay were investigated to fit equilibrium data. Three error functions of residual root mean square error, chi-square statistic, and average relative error were used to comfort us in the selected models, which were actually Dubinin-Radushkevich and Langmuir for BR2 and Frendlich, Temkin, and Halsay for MV.

Facile preparation of nitrogen and sulfur co-doped graphene-based aerogel for simultaneous removal of Cd2+ and organic dyes.

The need in simultaneous removal of heavy metals and organic compounds dictates the development of synthetic adsorbents with tailor-made properties. A nitrogen (N) and sulfur (S) co-doped graphene-based aerogel (GBA) modified with 2,5-dithiobisurea was synthesized hydrothermally for simultaneous adsorption of Cd2+ and organic dyes-safranin-O (SO), crystal violet (CV), and methylene blue (MB). 2,5-Dithiobisurea was used as nitrogen and sulfur sources to introduce N and S-containing functional group onto graphene oxide. The adsorption mechanism of GBA towards Cd2+ and organic dyes was studied by Dumwald-Wagner models and the results showed that surface and intraparticle diffusion was the key factor in controlling the rate of adsorption. The maximum adsorption capacities of GBA towards Cd2+, SO, CV, and MB comprised 1.755, 0.949, 0.538, and 0.389 mmol/g in monocomponent system, respectively. Adsorption synergism was observed with respect to Cd2+ in presence of the dyes. The performance of GBA with respect to Cd2+ removal from binary solutions, Cd2+-SO, Cd2+-CV, and Cd2+-MB, was enhanced by the presence of the dyes significantly, while the adsorption capacities towards the dyes were not affected by the presence of Cd2+.

A unique indolizinium alkaloid streptopertusacin A and bioactive bafilomycins from marine-derived Streptomyces sp. HZP-2216E.

Streptopertusacin A, a unique indolizinium alkaloid existing as a zwitterion, and six bafilomycins including two previously undescribed ones of 21,22-en-bafilomycin D and 21,22-en-9-hydroxybafilomycin D were isolated from a culture of the seaweed-derived Streptomyces sp. HZP-2216E. Structures of these isolated compounds were determined based on extensive NMR spectroscopic analyses, HRESIMS and MS-MS data. The stereochemical assignments were achieved by NOE information, chemical degradation, Marfey's method, and electronic circular dichroism (ECD) calculation. Streptopertusacin A is the first example of this type of indolizinium alkaloid from microorganisms and showed moderate activity against the growth of methicillin-resistant Staphylococcus aureus (MRSA). 21,22-en-bafilomycin D and 21,22-en-9-hydroxybafilomycin D had potent activities in inhibiting the proliferation of glioma cells and the growth of MRSA.

Ultrasound assisted adsorptive removal of hazardous dye Safranin O from aqueous solution using crosslinked graphene oxide-chitosan (GOCH) composite and optimization by response surface methodology (RSM) approach.

Chitosan (CH) was crosslinked with graphene oxide (GO) by combining solutions of CH and GO. Characterisations by ATR-FTIR, FE-SEM and XRD confirmed the formation of the GOCH composite. Removal of the dye Safranin Orange (SO) by ultrasonic adsorption from aqueous solution was tested by the composite. The removal of the cationic dye was more favourable at pH values greater than 5.2 and the optimum pH was found to be 6.5. The adsorption kinetics followed a pseudo-first order model and the rate-limiting step was identified as boundary layer diffusion from the Intraparticle diffusion model. The sonication assisted adsorption kinetic data were compared with the non-sonicated one and it was found that sonication has a marked effect on the adsorption kinetics. The Redlich Peterson adsorption isotherm described the adsorption with more resemblance to the Langmuir Model than the Freundlich Model suggesting that monolayer adsorption predominated. From Response Surface Methodology it was noted that the combined effect of pH and initial concentration was antagonistic while that of sonication time was synergistic. The optimum parameters from the RSM model were found to be pH 6.82, initial SO concentration 425mgL-1 and sonication time 25min. This was in good agreement with the experimental results.

Recent developments in the isolation, biological function, biosynthesis, and synthesis of phenazine natural products.

Phenazines are natural products which are produced by bacteria or by archaeal Methanosarcina species. The tricyclic ring system enables redox processes, which producing organisms use for oxidation of NADH or for the generation of reactive oxygen species (ROS), giving them advantages over other microorganisms. In this review we summarize the progress in the field since 2005 regarding the isolation of new phenazine natural products, new insights in their biological function, and particularly the now almost completely understood biosynthesis. The review is complemented by a description of new synthetic methods and total syntheses of phenazines.

New streptophenazines from marine Streptomyces sp. 182SMLY.

Six phenazines including three new ones were isolated from the culture of a marine actinomycete Streptomyces sp. 182SMLY. Based on the analyses of NMR, HRESIMS, optical rotation value, and CD data, the structures of these isolated compounds were determined as new phenazines of (-)-streptophenazines M-O and known phenazines of 1-carbomethoxyphenazine and (-)-streptophenazines A and B. (-)-Streptophenazine B showed activity in suppressing the growth of methicillin-resistant Staphylococcus aureus with MIC value of 4.2 µg/mL.

Isolation of Bioactive Phenazine-1-Carboxamide from the Soil Bacterium Pantoea agglomerans and Study of Its Anticancer Potency on Different Cancer Cell Lines.

The study was designed to investigate the anticancer effect of phenazine-1-carboxamide (PCN) isolated from the bacterium Pantoea agglomerans naturally present in soil. PCN showed cytotoxicity in a dose-dependent manner, and inhibitory concentrations on the cancer cell lines A549, HeLa, and SW480 were between 32 and 40 µM. Significantly increased concentrations of lactate dehydrogenase were found with increasing concentrations of PCN, which resulted in increased destruction of the cancer cell membrane. A significantly increased p53 level was accompanied by the increased production of cytochrome c protein in all cancer cell lines studied. This condition in cells leads to the overexpression of caspase 3 and Bcl-2 family proteins. Upregulation and downregulation of proapoptotic and antiproapoptotic proteins were analyzed for their messenger RNA and protein expression. The activation of caspases and their cleavage compounds paves the way for the complete apoptosis process in cancer cells. We conclude that P. agglomerans-derived PCN acts as an effective anticancer drug or compound.

Parametric and energy consumption optimization of Basic Red 2 removal by electrocoagulation/egg shell adsorption coupling using response surface methodology in a batch system.

In this study, response surface methodology (RSM) model was applied for optimization of Basic Red 2 (BR2) removal using electrocoagulation/eggshell (ES) coupling process in a batch system. Central composite design was used to evaluate the effects and interactions of process parameters including current density, reaction time, initial pH and ES dosage on the BR2 removal efficiency and energy consumption. The analysis of variance revealed high R(2) values (≥85%) indicating that the predictions of RSM models are adequately applicable for both responses. The optimum conditions when the dye removal efficiency of 93.18% and energy consumption of 0.840 kWh/kg were observed were 11.40 mA/cm(2) current density, 5 min and 3 s reaction time, 6.5 initial pH and 10.91 g/L ES dosage.

Heterocyclic Aromatic N-Oxidation in the Biosynthesis of Phenazine Antibiotics from Lysobacter antibioticus.

Heterocyclic aromatic N-oxides often have potent biological activities, but the mechanism for aromatic N-oxidation is unclear. Six phenazine antibiotics were isolated from Lysobacter antibioticus OH13. A 10 gene cluster was identified for phenazine biosynthesis. Mutation of LaPhzNO1 abolished all N-oxides, while non-oxides markedly increased. LaPhzNO1 is homologous to Baeyer-Villiger flavoproteins but was shown to catazlye phenazine N-oxidation. LaPhzNO1 and LaPhzS together converted phenazine 1,6-dicarboxylic acid to 1,6-dihydroxyphenazine N5,N10-dioxide. LaPhzNO1 also catalyzed N-oxidation of 8-hydroxyquinoline.

The Phenazine 2-Hydroxy-Phenazine-1-Carboxylic Acid Promotes Extracellular DNA Release and Has Broad Transcriptomic Consequences in Pseudomonas chlororaphis 30-84.

Enhanced production of 2-hydroxy-phenazine-1-carboxylic acid (2-OH-PCA) by the biological control strain Pseudomonas chlororaphis 30-84 derivative 30-84O* was shown previously to promote cell adhesion and alter the three-dimensional structure of surface-attached biofilms compared to the wild type. The current study demonstrates that production of 2-OH-PCA promotes the release of extracellular DNA, which is correlated with the production of structured biofilm matrix. Moreover, the essential role of the extracellular DNA in maintaining the mass and structure of the 30-84 biofilm matrix is demonstrated. To better understand the role of different phenazines in biofilm matrix production and gene expression, transcriptomic analyses were conducted comparing gene expression patterns of populations of wild type, 30-84O* and a derivative of 30-84 producing only PCA (30-84PCA) to a phenazine defective mutant (30-84ZN) when grown in static cultures. RNA-Seq analyses identified a group of 802 genes that were differentially expressed by the phenazine producing derivatives compared to 30-84ZN, including 240 genes shared by the two 2-OH-PCA producing derivatives, the wild type and 30-84O*. A gene cluster encoding a bacteriophage-derived pyocin and its lysis cassette was upregulated in 2-OH-PCA producing derivatives. A holin encoded in this gene cluster was found to contribute to the release of eDNA in 30-84 biofilm matrices, demonstrating that the influence of 2-OH-PCA on eDNA production is due in part to cell autolysis as a result of pyocin production and release. The results expand the current understanding of the functions different phenazines play in the survival of bacteria in biofilm-forming communities.

Enhanced production of phenazine-like metabolite produced by Streptomyces aurantiogriseus VSMGT1014 against rice pathogen, Rhizoctonia solani.

The efficacy of a rhizobacterium Streptomyces aurantiogriseus VSMGT1014 for the production of bioactive metabolites with antifungal properties was evaluated under in vitro conditions. The production of bioactive metabolites by S. aurantiogriseus VSMGT1014 in International Streptomyces Project-2 (ISP-2) broth, supplemented with glucose and ammonium acetate was found to be the most suitable carbon and nitrogen sources for the maximum production of bioactive metabolites against rice pathogen, Rhizoctonia solani. The zone of inhibition range from 23.5 to 28.5 mm and 10.3 to 18.3 mm for glucose and ammonium acetate supplemented media, respectively. The culture filtrate of S. aurantiogriseus VSMGT1014 at pH 7.5, 37 °C at 120 rpm in 6 days of incubation showed the maximum production of bioactive metabolites with antagonistic potential. The crude metabolite was characterized by different spectral studies such as Ultraviolet spectrum, infrared-spectrum and based on the different analytical techniques, including thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) with the retention time 29.4 and the bioactive metabolite was identified as phenazine, which was confirmed by pure phenazine compound as positive control.

New phenazine analogues from Streptomyces sp. IFM 11694 with TRAIL resistance-overcoming activities.

Two new phenazine derivatives, aotaphenazine (1) and 5,10-dihydrophencomycin (2), were isolated from the ethyl acetate extract of Streptomyces sp. IFM 11694. In addition, the known 1-phenazinecarboxylic acid (3), phencomycin (4) and 1,6-phenazinedicarboxylic acid (5) were identified. The structures of the isolated compounds (1-5) were characterized by spectroscopic methods including NMR and mass spectrometry data. Compound 1 showed the ability to overcome tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) resistance at concentration of 12.5 µM. Aotaphenazine (1) enhanced the levels of apoptosis inducing proteins DR4, DR5, p53 and also decreased the levels of cell survival protein Bcl-2 in TRAIL-resistant human gastric adenocarcinoma (AGS) cells in a dose-dependent manner.

Pontemazines A and B, phenazine derivatives containing a methylamine linkage from Streptomyces sp. UT1123 and their protective effect to HT-22 neuronal cells.

New phenazine derivatives with a methylamine linker, Pontemazines A (1) and B (2), were isolated from the culture broth of Streptomyces sp. UT1123. The structures of compounds 1 and 2 were determined by NMR spectroscopy and high-resolution mass spectrometry. These compounds consist of a 9-mehoxyphenazine connected to a benzamide functional group by a unique methylamine linker instead of the more common methyl ether. Pontemazines A and B possess a neuronal cell protective effect on glutamate-induced mouse hippocampal HT-22 cell damage.

Synthesis, characterization, swelling and dye adsorption properties of starch incorporated acrylic gels.

Several hydrogels were prepared by a free radical polymerization of acrylic acid (AA), sodium acrylate (SA) and AA/hydroxy ethyl methacrylate (HEMA) in the presence of starch in water. These starch incorporated acrylic gels were prepared by varying the concentration of the initiator, monomer, crosslinker and the starch. The resulting gels were characterized by FTIR, SEM, XRD, DTA-TGA, pH at point zero charge (PZC), swelling and the diffusion in water. The gels showed high adsorption and removal% of Safranine T (ST) and Brilliant Cresyl Blue (BCB) dyes from water. The swelling and the adsorption data were fitted to different kinetic models and isotherms. Amongst the three kinds of gels, the starch incorporated sodium polyacrylate gel showed the highest adsorption of 9.7-85.3mg/L (97-61% removal) of BCB dye and 9.1-83mg/L (91-60% removal) of ST dye for a feed dye concentration of 10-140mg/L.

[Isolation, identification and characterization of rice rhizobacterium Pseudomonas aeruginosa PA1201 producing high level of biopesticide "Shenqinmycin" and phenazine-1-carboxamide].

OBJECTIVE: To identify bacterial strains with the inhibitory activity to rice pathogens, and to evaluate their potentials for the development of new biopesticides. METHODS: Rice rhizosphere Pseudomonas strains were isolated using 1-aminocyclopropane-1-carboxylic acid as the sole carbon source. Strain PA1201 was further identified through morphological analysis, biochemical characterization, 16S rDNA sequence analysis and phospholipid fatty acid profiling. Qualitative and quantitative analysis of the production of the green pesticide Shenqinmycin as well as phenazine-1-carboxamide produced by PA1201 was done by HPLC. Cytotoxicity of PA1201 was evaluated using human alveolar epithelial cell line A549 and Drosophila melanogaster as hosts. RESULTS: Strain PA1201 inhibited Rhizotonia solani Kuhn and Xanthomonas oryzae pv. oryzae, the causal agents of rice sheath blight and bacterial blight, respectively. It was further identified as Pseudomonas aeruginosa PA1201, which produces shenqinmycin and phenazine-1-carboxamide. The fermentation titer of shenqinmycin and phenazine-1-carboxamide in the PPM medium was 81.7 mg/L and 18. 1 mg/L, respectively. In the medium supplemented with soybean meal and corn steep liquor, the level of shenqinmycin and phenazine-1-carboxamide reached 926. 9 mg/L and 489. 5 mg/L. PA1201 also produced high level of extracellular protease and was toxic to human cell line and fruit fly. CONCLUSION: Strain PA1201 could be engineered for higher yield of Shenqinmycin or for a new biopesticide.

Response surface methodology approach for optimization of simultaneous dye and metal ion ultrasound-assisted adsorption onto Mn doped Fe3O4-NPs loaded on AC: kinetic and isothermal studies.

In the present work, the usefulness of ultrasonic power as a dispersion and mixing tool to accelerate the adsorption of Safranin O (SO), methylene blue (MB), Pb(2+) ions and Cr(3+) ions onto the novel composite Fe3O4-NPs-AC adsorbent was investigated. This new material was extensively characterized and analyzed by different techniques such as XRD, FESEM, Raman spectroscopy and FT-IR. Central composite design (CCD) based on designed runs revealed that adsorbent mass, sonication time, MB concentration, SO concentration, Pb(2+) ion and Cr(3+) ion concentration and some of their interactions have significant contributions to the target compounds removal percentages. A combination of response surface methodology and Design-Expert software was used to qualify and estimate the influence and magnitude of each terms contribution to the response. An optimization study using the following investigated increments of the effective variables, adsorbent mass (0.01-0.03 g), sonication time (2-6 min), initial dye concentration (5-25 mg L(-1)), and initial metal ion concentration (20-60 mg L(-1)), revealed that fixing the experimental variables at 0.025 g of Mn-Fe3O4-NPs-AC, with a 3 min sonication time, and 20 mg L(-1) of MB, 10 mg L(-1) of SO, 38 mg L(-1) of Pb(2+) ions and 42 mg L(-1) of Cr(3+) ions at room temperature lead to the achievement of the best characteristics and performance. Conduction of 32 experiments according to the limitations of CCD and a subsequent analysis of variance (ANOVA) gave useful information about the significant and also approximate contributions of each term (main and interaction of variables) in an empirical equation for the expected response. The results indicate that the R(2) values are more than 0.988 and the adjusted R(2) values are in reasonable agreement with R(2). Under the optimal conditions, the MB, SO, Pb(2+) ion and Cr(3+) ion removal efficiencies reached 99.54%, 98.87%, 80.25% and 99.54% after 3 min, while their equilibrium data with high performance can be represented by Langmuir isotherms and a pseudo second-order kinetic model. The maximum adsorption capacities for the single component system, 229.4 mg g(-1) for MB, 159.7 mg g(-1) for SO, 139.5 mg g(-1) for Pb(2+) ions and 267.4 mg g(-1) for Cr(3+) ions, support the high efficiency of Mn-Fe3O4-NPs-AC as a new adsorbent.