Co-factor independent oxidases ncnN and actVA-3 are involved in the dimerization of benzoisochromanequinone antibiotics in naphthocyclinone and actinorhodin biosynthesis.

Streptomyces produce complex bioactive secondary metabolites with remarkable chemical diversity. Benzoisochromanequinone polyketides actinorhodin and naphthocyclinone are formed through dimerization of half-molecules via single or double carbon-carbon bonds, respectively. Here we sequenced the genome of S. arenae DSM40737 to identify the naphthocyclinone gene cluster and established heterologous production in S. albus J1074 by utilizing direct cluster capture techniques. Comparative sequence analysis uncovered ncnN and ncnM gene products as putative enzymes responsible for dimerization. Inactivation of ncnN that is homologous to atypical co-factor independent oxidases resulted in the accumulation of fogacin, which is likely a reduced shunt product of the true substrate for naphthocyclinone dimerization. In agreement, inactivation of the homologous actVA-3 in S. coelicolor M145 also led to significantly reduced production of actinorhodin. Previous work has identified the NAD(P)H-dependent reductase ActVA-4 as the key enzyme in actinorhodin dimerization, but surprisingly inactivation of the homologous ncnM did not abolish naphthocyclinone formation and the mutation may have been complemented by an endogenous gene product. Our data suggests that dimerization of benzoisochromanequinone polyketides require two-component reductase-oxidase systems.

Enhanced sunlight photocatalytic activity and biosafety of marine-driven synthesized cerium oxide nanoparticles.

This contribution presents the biosynthesis, physiochemical properties, toxicity and photocatalytic activity of biogenic CeO2 NPs using, for the first time, marine oyster extract as an effective and rich source of bioreducing and capping/stabilizing agents in a one-pot recipe. CeO2 NPs formation was initially confirmed through the color change from light green to pale yellow and subsequently, their corresponding absorption peak was spectroscopically determined at 310 nm with an optical band-gap of 4.67 eV using the DR-UV technique. Further, XRD and Raman analyses indicated that nanoceria possessed face-centered cubic arrangements without any impurities, having an average crystallite size of 10 nm. TEM and SEM results revealed that biogenic CeO2 NPs was approximately spherical in shape with a median particle size of 15 ± 1 nm. The presence of various bioorganic substances on the surface of nanoparticles was deduced by FTIR and TGA results. It is found that marine-based nanoceria shows no cytotoxic effect on the normal cell, thus indicating their enhanced biocompatibility and biosafety to living organisms. Environmentally, due to energy band gap, visible light-activated CeO2 nanocatalyst revealed superior photocatalytic performance on degradation of methylene blue pollutant with removal rate of 99%. Owing to the simplicity, cost-effectiveness, and environmentally friendly nature, this novel marine biosynthetic route paves the way for prospective applications of nanoparticles in various areas.

Synergistic effects of combinatorial chitosan and polyphenol biomolecules on enhanced antibacterial activity of biofunctionalaized silver nanoparticles.

The present study reports the synergistic antibacterial activity of biosynthesized silver nanoparticles (AgNPs) with the aid of a combination of chitosan and seaweed-derived polyphenols as a green synthetic route. Under optimum synthesis conditions, the rapid color change from yellowish to dark brown and UV-visible absorption peak at 425 confirmed the initial formation of AgNPs. DLS, TEM, XRD, and EDX analyses revealed the spherical shape of pure biogenic AgNPs with a mean diameter size of 12 nm ± 1.5 nm, and a face-centered cubic crystal structure, respectively. FTIR and TGA results indicated the significant contribution of chitosan and polyphenol components into silver ions bioreduction and thermal stability of freshly formed AgNPs. Long-term colloidal stability of AgNPs was obtained after 6-month storage at room temperature. The bio-prepared AgNPs possessed a negative surface charge with a zeta potential value of - 27 mV. In contrast to naked chemical silver nanoparticles, the green Ag nanosamples demonstrated the distinct synergistic antibacterial in vitro toward all selected human pathogens presumably due to the presence of high content of biomolecules on their surface. The results show that synergy between chitosan and polyphenol results in the enhancement of bactericidal properties of biogenic AgNPs. We also highlighted the underlying mechanism involved in AgNPs formation based on nucleophile-electrophile interaction.

Genotyping-Guided Discovery of Persiamycin A From Sponge-Associated Halophilic Streptomonospora sp. PA3.

Microbial natural products have been a cornerstone of the pharmaceutical industry, but the supply of novel bioactive secondary metabolites has diminished due to extensive exploration of the most easily accessible sources, namely terrestrial Streptomyces species. The Persian Gulf is a unique habitat for marine sponges, which contain diverse communities of microorganisms including marine Actinobacteria. These exotic ecosystems may cradle rare actinomycetes with high potential to produce novel secondary metabolites. In this study, we harvested 12 different species of sponges from two locations in the Persian Gulf and isolated 45 symbiotic actinomycetes to assess their biodiversity and sponge-microbe relationships. The isolates were classified into Nocardiopsis (24 isolates), Streptomyces (17 isolates) and rare genera (4 isolates) by 16S rRNA sequencing. Antibiotic activity tests revealed that culture extracts from half of the isolates displayed growth inhibitory effects against seven pathogenic bacteria. Next, we identified five strains with the genetic potential to produce aromatic polyketides by genotyping ketosynthase genes responsible for synthesis of carbon scaffolds. The combined data led us to focus on Streptomonospora sp. PA3, since the genus has rarely been examined for its capacity to produce secondary metabolites. Analysis of culture extracts led to the discovery of a new bioactive aromatic polyketide denoted persiamycin A and 1-hydroxy-4-methoxy-2-naphthoic acid. The genome harbored seven gene clusters involved in secondary metabolism, including a tetracenomycin-type polyketide synthase pathway likely involved in persiamycin formation. The work demonstrates the use of multivariate data and underexplored ecological niches to guide the drug discovery process for antibiotics and anticancer agents.

Interactions of oxidative DNA damage and CYP1A gene expression with the liver enzymes in Klunzinger's mullet exposed to benzo[a]pyrene.

Benzo[a]pyrene (B[a]P) is an important contaminant whose liver biotransformation is dependent on the species, the route of exposure and the concentration. The goal of this study was to assess the interactions of oxidative DNA damage and CYP1A gene expression with the liver enzymes in Klunzinger's mullet (Liza klunzingeri) exposed to benzo[a]pyrene. Sublethal doses of B[a]P (5, 10 and 50 mg/kg) were intraperitoneally administered to the fish for 14 days. The alterations in antioxidant enzymes' activity (SOD, CAT, and GPX), hepatic enzymes' activity (ALT, AST and ALP), DNA damage (measured by comet assay and cellProfiler software) and CYP1A gene expression in the fish liver were studied on the 1st, 3rd, 7th and 14th days. The determination of these parameters in the liver showed that most of these parameters significantly increased mostly in a time-dependent manner. Multiple regression analysis showed that DNA damage and CYP1A gene expression had positive correlations with the liver enzymes in this fish species intraperitoneally exposed to these concentrations. Moreover, these interactions indicated that theses parameters are sensitive biomarkers for the exposure to B[a]P in Klunzinger's mullet. However, other possible factors and B[a]P metabolites should be considered in future studies for better elucidating the biotransformation mechanisms and introducing better biomarkers of B[a]P.

Short-term induction of vitellogenesis in the immature male yellowfin seabream ( Acanthopagrus latus) exposed to bisphenol A and 17 ß-estradiol.

Bisphenol A (BPA) is a known environmental endocrine-disrupting chemical that is widely used in plastics manufacturing. BPA enters in the aquatic environment mainly through urban and industrial sewage effluents, thereby posing a potential threat to organisms living in these ecosystems. This study was conducted to investigate the effect of BPA on VTG production with direct (sodium dodecyl sulfate-polyarylamide gel electrophoresis) and indirect (alkali-labile phosphate (ALP), total plasma calcium and protein) methods in immature male yellowfin seabream ( Acanthopagrus latus) as a marine fish model. Fish were randomly distributed into seven groups that were administered 1, 10, 50, and 100 µg g-1 week-1 of BPA and 2 µg g-1week-1 of 17ß-estradiol (E2) over a period of 2 weeks. Solvent controls received olive oil, whereas controls were not injected. The fish were sampled on days 0, 7, and 14, and their blood plasma and liver were obtained. The results showed that the hepatosomatic index of all treated fish was elevated in comparison with controls. Direct and indirect indicators showed that fish VTG protein was induced by BPA and E2 exposure. The protein was found to have two bands with molecular weights around 210 and 190 KDa. ALP, total plasma calcium and protein levels were increased in dose- and time-dependent manners. The results of this study demonstrated that short-term exposure of yellowfin seabream to BPA induced adverse effects in the reproductive system of hermaphrodite fish.

Common features of tuberculosis and sarcoidosis.

Tuberculosis (TB) is a disease caused by Mycobacterium tuberculosis. Despite the availability of novel therapeutic approaches, TB is considered as one of the leading causes of death due to infectious diseases worldwide. Alveolar macrophages are the first line of defense against M. tuberculosis; they ingest and sequester the bacilli within granulomatous structures. Control and resolution of the infection requires activated T lymphocytes as well as Th1 cytokines. There are two forms of TB: active TB and latent TB. Latent TB is a state in which M. tuberculosis survives in the body without causing overt signs and symptoms. People with latent TB are noncontagious. However, M. tuberculosis can become active in the body, multiply, and cause overt TB. Sarcoidosis, on the other hand, is an autoimmune disease of unknown etiology which can affect multiple systems of the body. Nonspecific constitutional symptoms, such as fever, fatigue, malaise, and weight loss, are present in approximately one-third of patients. Chest X-ray usually shows hilar and mediastinal lymphadenopathy. Although the lungs are the most common sites of inflammation, sarcoidosis can also involve other organs, such as the eyes (intraocular and adnexal), skin, lymph nodes, salivary glands, heart, spleen, liver, and the nervous system. Recent investigations have provided further insights into the genetic basis of sarcoidosis and the way genotype determines the clinical presentation and phenotype of patients. Histopathologic features are usually insufficient for diagnosis of sarcoidosis. Diagnosis of sarcoidosis in endemic areas for TB can become a great challenge. Both TB and sarcoidosis are granulomatous diseases; TB is characterized by caseating granulomas, whereas sarcoidosis is characterized by noncaseating granulomas. New cases of sarcoidosis are increasingly being diagnosed in areas endemic for TB due to increased orientation of physicians and availability of diagnostic modalities. However, it is often difficult to differentiate sarcoidosis from TB, especially when caseous necrosis is not seen and acid-fast staining is negative in the biopsy specimen of patient with TB. Granulomatous inflammation in sarcoidosis is believed to be caused by the presence of a persistent poorly degradable unknown antigen in combination with a nonresolving host response. M. tuberculosis has been extensively studied as a possible cause of sarcoidosis. Results suggest that granulomas form in the lungs as a result of the immune response to inhaled M. tuberculosis and serve as the central site of host-pathogen interaction during M. tuberculosis infection. M. tuberculosis DNA detection in sarcoidosis samples by traditional polymerase chain reaction (PCR) has been used for the pathological study of sarcoidosis; however, it is likely that real time quantitative PCR analysis of specific mRNAs and microRNAs will be necessary as a sensitive, precise, and rapid diagnostic test for detecting trace of TB in Sarcoidosis. In conclusion, diagnosis of sarcoidosis in areas with a high burden of TB poses a significant challenge. Improved diagnostic tests including genetic tests can improve our knowledge and help in distinguishing these two diseases.

Physicochemical study of a novel chimeric chitinase with enhanced binding ability.

Chitinases are slow-reacting but important enzymes as they are anticipated to have diverse applications. The role of a chitin-binding domain (ChBD) in enhancing the quality of binding is essential information for purposeful engineering of chitinases. The idea of making hybrid chitinases by fusing a known ChBD to a chitinase, which naturally lacks ChBD is of interest especially for bio-controlling purposes. Therefore, in the present study, the ChBD of Serratia marcescens chitinase B was selected and fused to the fungal chitinase, Trichoderma atroviride Chit42. Both Chit42 and chemric Chit42 (ChC) showed similar activity towards colloidal chitin with specificity constants of 0.83 and 1.07 min(-1), respectively, same optimum temperatures (40°C), and similar optimum pH (4 and 4.5, respectively). In the presence of insoluble chitin, ChC showed higher activity (70%) and obtained a remarkably higher binding constant (700 times). Spectroscopic studies indicated that chimerization of Chit42 caused some structural changes, which resulted in a reduction of α-helix in ChC structure. Chemical and thermal stability studies suggested that ChC had a more stable structure than Chit42. Hill analysis of the binding data revealed mixed-cooperativity with positive cooperativity governing at ChC concentrations below 0.5 and above 2 µM in the presence of insoluble chitin. It is suggested that the addition of the ChBD to Chit42 affords structural changes which enhance the binding ability of ChC to insoluble chitin, improving its catalytic efficiency and increasing its thermal and chemical stability.

Designing a new chitinase with more chitin binding and antifungal activity.

Chitinases have the ability of chitin digestion that constitutes a main compound of the cell wall in many of the phytopathogens such as fungi. Chitinase Chit42 from Trichoderma atroviride PTCC5220 is considered to play an important role in the biocontrol activity of this fungus against plant pathogens. Chit42 lacks a chitin binding domain (ChBD). We have produced a chimeric chitinase with stronger chitin-binding capacity by fusing to Chit42 a ChBD from Serratia marcescens Chitinase B. The fusion of ChBD improved the affinity to crystalline and colloidal chitin and also the enzyme activity of the chimeric chitinase when compared with the native Chit42. The chimeric chitinase showed higher antifungal activity toward phytopathogenic fungi.