Schultzthygesen9325

This immune regulation was related to the NF-κB and MAPK pathways in RAW264.7 cells.A fosmid metagenomic library containing 9.7 × 104 clones was constructed. A novel esterase, XtjR8, was isolated through functional screening. XtjR8 shared the maximum amino acid identity (44%) with acetyl-hydrolase from Streptomyces hygroscopicus, and was classified into family IV esterase. XtjR8 exhibited the highest hydrolytic activity for p-nitrophenyl acetate at 40 °C and pH 8.0, and presented more than 40% activity from 20 °C to 80 °C. More importantly, XtjR8 displayed the ability to hydrolyze both phthalate monoesters and diesters, this feature is extremely rare among previously reported esterases. Site-directed mutagenesis experiments revealed that the catalytic triad residues were Ser152, Glu246, and His276. Among them, Ser152 formed a hydrogen bond with dibutyl phthalate (DBP) by molecular docking, Gly84, Gly85, and Leu248 of conserved motifs formed hydrophobic interactions with DBP, respectively, which were important for the catalytic activity. Considering its wide range of temperature and hydrolytic potential toward phthalate esters, XtjR8 will be served as an interesting candidate for biodegradation and industrial applications.Fungi produce several toxins active against plants, animal or humans. Among them, ribotoxins are enzymes that specifically attack ribosomes irreparably compromising protein synthesis, useful as insecticides or as anticancer agents. Here, a novel ribotoxin from the edible mushroom Pleurotus ostreatus has been purified and characterized. This ribotoxin, named Ostreatin, is a specific ribonuclease releasing α-fragment when incubated with yeast or rabbit ribosomes. Ostreatin shows IC50 of 234 pM in rabbit reticulocyte lysate, and metal dependent endonuclease activity. Following the completion of Ostreatin primary structure, we ascertained that this toxin is homologous to Ageritin, the first ribotoxin-like protein from the basidiomycete Agrocybe aegerita, with which it shares 38.8% amino acid sequence identity. Ostreatin consists of 131 amino acid residues with an experimental molecular mass of 14,263.51 Da ([M+H+]+). Homology modeling revealed that Ostreatin and Ageritin share a similar fold in which the common catalytic triad is conserved. Purified Ostreatin lacks N-terminal and C-terminal peptides, which instead are present in the Ostreatin coding sequence. Such peptides are probably involved in protein sorting and for this they could be removed. Our findings confirm the presence of ribotoxin-like proteins in basidiomycetes edible mushrooms, that we propose as novel tool for biotechnological applications.Over the decades, several nanoparticles have been developed for biomedical applications, still facile green synthesis derived nanoparticles showed tremendous attraction due to avoid of toxic solvent, ease of synthesis and low cost. Here, facile one pot in situ green synthesis is reported to develop silver nanoparticles with the aid of natural polysaccharide presented in sweet lemon peel waste derived carbon dot (CD) acted as a reducing and stabilizing agent at room temperature. The synthesis of CD and CD based silver nanoparticles (CD@AgNPs) was characterized by FTIR, UV-vis spectroscopy, fluorescence spectrophotometer, XRD and TEM. CD@AgNPs exhibited excellent antimicrobial activity against E. coli at very low concentration of 5.0 μg/ml. Interestingly, CD showed selective cytotoxicity against MCF7 breast cancer cells with the IC50 of 10 μg/ml while CD@AgNPs demonstrated synergistic effect on cytotoxicity. It is found that the cells death of MCF7 cells mainly occurred through the up-regulation of intracellular reactive oxygen species (ROS). Therefore, the synthesized CD@AgNPs may show an efficient anticancer agent for targeted breast cancer therapy in future.Innate immunity driven by pattern recognition receptor (PRR) protects the host from invading pathogens. Aquatic animals like fish where the adaptive immunity is poorly developed majorly rely on their innate immunity modulated by PRRs like toll-like receptors (TLR) and NOD-like receptors (NLR). However, current development to improve the fish immunity via TLR/NLR signaling is affected by a poor understanding of its mechanistic and structural features. This review discusses the structure of fish TLRs/NLRs and its interaction with pathogen associated molecular patterns (PAMPs) and downstream signaling molecules. Over the past one decade, significant progress has been done in studying the structure of TLRs/NLRs in higher eukaryotes; however, structural studies on fish innate immune receptors are undermined. Several novel TLR genes are identified in fish that are absent in higher eukaryotes, but the function is still poorly understood. Unlike the fundamental progress achieved in developing antagonist/agonist to modulate human innate immunity, analogous studies in fish are nearly lacking due to structural inadequacy. This underlies the importance of exploring the structural and mechanistic details of fish TLRs/NLRs at an atomic and molecular level. This review outlined the mechanistic and structural basis of fish TLR and NLR activation.The intestinal epithelium is known as an important barrier to protect the body from harmful pathogens or toxic substance that may induce intestinal barrier injury. The aim of this study was to investigate the effects of polysaccharide from the seeds of Plantago asiatica L. (PLP) on nonylphenol (NP) induced intestinal barrier injury in vitro. Caco-2 cells were pretreated with PLP, or co-cultured with PLP and NP simultaneously, and cytotoxicity, LDH leakage, transepithelial electrical resistance (TEER), FITC-dextran flux and tight junction (TJ) proteins were conducted to evaluate the intestinal barrier function. DJ4 mouse The results suggested that PLP pretreatment or co-culture with NP could significantly attenuated NP induced Caco-2 cytotoxicity, suppressed LDH release, restored the TEER value and paracellular permeability of Caco-2 monolayers, which were attributed to enhancing the TJ protein expressions. In addition, PLP co-cultured with NP possessed better protective effects against NP induced cytotoxicity. This study indicated that PLP assuaged NP induced intestinal barrier injury by increasing TJ, and threw light on the development of a dietary supplementation for preventing exogenous toxic substances induced intestinal barrier injury or improving intestinal TJ barrier function.