DataSheet1_Synergistic Effect of Proteinase Activity by Purification and Identification of Toxic Protease From Nemopilema nomurai.docx

Scyphozoan Nemopilema nomurai envenomation is an unresolved threat to human health in Asian waters. Nemopilema nomurai venom metalloproteinases show important toxicities in skin damage and inflammation, but there is still no purified protein for further studies. In this study, high proteinase activity fractions in tentacle autolysis were isolated by ammonium sulfate precipitation, DEAE Sepharose Fast Flow, and Superdex 75 chromatography successively. Purification was guided by azocasein hydrolysis activity and SDS-PAGE. The final products were analyzed by LC-MS/MS. Four elution peaks purified by Superdex 75 chromatography had multiple protein bands but did not show proteinase activity. These fractions would recover proteinase activity after mixing again. Regulation mechanisms were speculated as binding metalloproteinase regulator or disaggregating metalloproteinase inhibitor by LC-MS/MS analysis. For the first time, a synergistic effect in N. nomurai proteinase activity was found in the purification process.</p

Identification of fungal strains isolated from the jellyfish samples based on morphological characteristics as well as DNA analysis of the internal transcribed spacer (ITS) region.

<p>The closest relatives in GenBank according toBLAST search were presented.</p

HPLC chromatograms of EtOAc extracts of (A) <i>Tilletiopsis</i> sp., (B)<i>Tilletiopsis albescens</i>, (C) <i>P</i>. <i>lilacinum</i>, (D) <i>Cladosporium</i> sp. cultivated with WMB, RM, ISP2 media detected at 210nm or 220nm.

<p>HPLC chromatograms of EtOAc extracts of (A) <i>Tilletiopsis</i> sp., (B)<i>Tilletiopsis albescens</i>, (C) <i>P</i>. <i>lilacinum</i>, (D) <i>Cladosporium</i> sp. cultivated with WMB, RM, ISP2 media detected at 210nm or 220nm.</p

Synthesis and Characterization of Slow-Release Chitosan Oligosaccharide Pyridine Schiff Base Copper Complexes with Antifungal Activity

Herein, a series of chitosan oligosaccharide copper complexes modified with pyridine groups (CPSx-Cu complexes) were successfully prepared via the Schiff base reaction and ion complexation reaction for slow-release fungicide. The structures of the synthesized derivatives were characterized via Fourier transform infrared spectroscopy and 1H and 13C nuclear magnetic resonance spectroscopy, and the unit configuration of the complexes was calculated using Gaussian software. The slow-release performance experiment demonstrated that the cumulative copper ion release rate of CPSx-Cu complexes was dependent on the type of substituents on the pyridine ring. Furthermore, the in vitro and in vivo antifungal activities of the CPSx-Cu complexes were investigated. At a concentration of 0.4 mg/mL, CPSx-Cu complexes completely inhibited the growth of Pythium vexans and Phytophthora capsici. Results indicated that CPSx-Cu complexes with slow-release ability exhibited better antifungal activity than thiodiazole-copper and copper sulfate basic. This study confirmed that combining chitosan oligosaccharide with bioactive pyridine groups and copper ions is an effective approach to further developing slow-release copper fungicides, providing new possibilities for the application of copper fungicides in green agriculture. This study lays the foundation for further studies on biogreen copper fungicides

Exploring the Antibacterial and Antifungal Potential of Jellyfish-Associated Marine Fungi by Cultivation-Dependent Approaches

<div><p>Fungi isolated from marine invertebrates are of considerable importance as new promising sources of unique secondary metabolites with significant biomedical potential. However, the cultivable fungal community harbored in jellyfish was less investigated. In this work, we seek to recover symbiotic fungi from different tissues of jellyfish <i>Nemopilema nomurai</i>. A total of seven morphotypes were isolated, which were assigned into four genera (<i>Aspergillus</i>, <i>Cladosporium</i>, <i>Purpureocillium</i>, and <i>Tilletiopsis</i>) from two phyla (Ascomycota and Basidiomycota) by comparing the rDNA-ITS sequences with the reference sequences in GenBank. The most fungi were found in the inner tissues of subumbrella. Two of the cultivation-independent procedures, changing media type and co-cultivation, were employed to maximize the complexity of metabolites. Thus, thirteen EtOAc gum were obtained and fingerprinted by High Performance Liquid Chromatography (HPLC) equipped with a photodiode array (PDA) detector. Antibacterial and antifungal activities of these complex mixtures were tested against a panel of bacterial and fungal pathogens. The antimicrobial results showed that all of the 13 EtOAc extracts displayed different levels of antibacterial activity, three of which exhibited strong to significant antibacterial activity to the bacterial pathogens <i>Staphylococcus aureus</i> and <i>Salmonella entrica</i>. Antifungal activity indicated that the EtOAc extracts from pure culture of <i>Aspergillus versicolor</i> and co-culture of <i>A</i>. <i>versicolor</i> and <i>Tilletiopsis</i> sp. in rice media were promising for searching new compounds, with the maximal mycelial growth inhibition of 82.32% ± 0.61% for <i>Rhizoctonia solani</i> and 48.41% ± 11.02% for <i>Botrytis cinerea</i> at 200 μg/ml, respectively. This study is the first report on the antibacterial and antifungal activity of jellyfish-associated fungi and allows the first sight into cultivable fungal community residing in jellyfish. Induced metabolites by cultivation-dependent approaches provides a new reservoir for drug discovery from jellyfish-derived fungi.</p></div

Results of antifungal activity of crude extracts from pure stain SmT07 and co-culture strains in liquid Wickerham’ medium and solid rice medium against three common phytopathogenic fungi.

<p>Bars indicate the standard deviation from four replicates. Different alphabets above the bars have mean values that are significantly different (p < 0.05).</p

Neighbor-joining phylogenetic tree from analysis of available rDNA-ITS sequences of all identified jellyfish-derived fungi.

<p>The ITS sequences obtained in this work were indicated in bold. Bootstrap values were indicated at nodes of each branch based on a neighbor-joining analysis of 1000 replicates. Only values of > 50% were shown. Scale bar was equal to 0.1 substitutions per nucleotide position.</p

Results of agar diffusion assays for the antibiotic activity of ethyl acetate (EtOAc) extracts from OSMAC cultures of the giant jellyfish-derived fungi.

<p>Results of agar diffusion assays for the antibiotic activity of ethyl acetate (EtOAc) extracts from OSMAC cultures of the giant jellyfish-derived fungi.</p

Facile Template-Free Fabrication of Hollow Nestlike α‑Fe₂O₃ Nanostructures for Water Treatment

Hollow nestlike α-Fe₂O₃ spheres were successfully synthesized via a facile template-free, glycerol-mediated hydrothermal process employing microwave heating. The product was characterized using X-ray diffraction, field-emission scanning electron microscopy, transmission electron microscopy, and nitrogen adsorption measurements. The as-prepared product was found to consist of hierarchically nanostructured spheres assembled of nanorod subunits. The effect of the relative amount of glycerol in the reaction system on the composition and morphology of the products was systematically studied, and a possible formation mechanism of the hollow nestlike spheres was proposed. Because of their large surface area and unique mesoporous structure, we investigated the potential application of the hollow α-Fe₂O₃ spheres in water treatment. With maximum removal capacities of 75.3, 58.5, and 160 mg g^–1 for As­(V), Cr­(VI), and Congo red, respectively, these novel nanostructures have the potential to be used as low-cost and efficient adsorbent materials for the removal of toxic metal ions and organic pollutants from water