0000000000099655

AUTHOR

T. Faddetta

The microbial community of the coelomic fluid of the sea urchin Paracentrotus lividus

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Metabolic pathways in Microbispora sp. ATCC-PTA 5024, producer of NAI-107 lantibiotic

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Bioactive molecules from soil and marine bacteria: new potential applications

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Elucidating the molecular physiology of lantibiotic NAI-107 production in Microbispora ATCC-PTA-5024.

Background The filamentous actinomycete Microbispora ATCC-PTA-5024 produces the lantibiotic NAI-107, which is an antibiotic peptide effective against multidrug-resistant Gram-positive bacteria. In actinomycetes, antibiotic production is often associated with a physiological differentiation program controlled by a complex regulatory and metabolic network that may be elucidated by the integration of genomic, proteomic and bioinformatic tools. Accordingly, an extensive evaluation of the proteomic changes associated with NAI-107 production was performed on Microbispora ATCC-PTA-5024 by combining two-dimensional difference in gel electrophoresis, mass spectrometry and gene ontology approaches. R…

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Novel Sortase A Inhibitors to Counteract Gram-Positive Bacterial Biofilms

Sortase A (SrtA) is a membrane enzyme responsible for the covalent anchoring of surface proteins on the cell wall of Gram-positive bacteria. Nowadays it is considered an interesting target for the development of new anti-infective drugs which aim to interfere with important Gram-positive virulence mechanisms. Along the years, we studied the anti-staphylococcal and anti-biofilm activity of some natural and synthetic polyhalogenated pyrrolic compounds, called pyrrolomycins. Some of them were active on Gram-positive pathogens at a μg/mL range of concentration (1.5-0.045 μg/mL) and showed a biofilm inhibition in the range of 50-80%. [1-3] In light of these encouraging results, herein we present…

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Streptomyces coelicolor Vesicles: Many Molecules To Be Delivered

ABSTRACT Streptomyces coelicolor is a model organism for the study of Streptomyces, a genus of Gram-positive bacteria that undergoes a complex life cycle and produces a broad repertoire of bioactive metabolites and extracellular enzymes. This study investigated the production and characterization of membrane vesicles (MVs) in liquid cultures of S. coelicolor M145 from a structural and biochemical point of view; this was achieved by combining microscopic, physical and -omics analyses. Two main populations of MVs, with different sizes and cargos, were isolated and purified. S. coelicolor MV cargo was determined to be complex, containing different kinds of proteins and metabolites. In particul…

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Characterization of the microbiota from coelomic fluid of the sea urchin Paracentrotus lividus

The study of the microbiota is a subject of considerable and growing interest since it is drawing new important perspectives in the life sciences concerning the functional relationships between metazoans and microbial cells. In fact, it has already shown that the endogenous microbial community affects various physiological activities of multicellular organisms. The coelomic cavity of echinoderms contains a fluid in which coelomocytes are reported to exert immune functions like phagocytosis, opsonization and production of antimicrobial agents against marine bacteria [1, 2]. However, up to day nothing is known about the endogenous bacterial population of coelomic fluid. We focused on this iss…

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Thymus vulgaris Essential Oil and Hydro-Alcoholic Solutions to Counteract Wooden Artwork Microbial Colonization

Aromatic plants represent a source of natural products with medicinal properties, and are also utilized in the food and pharmaceutical industries. Recently, the need for eco-compatible and non-toxic products, safe for both the environment and human health, have been proposed for the sustainable conservation of historic–artistic artifacts. In this study, in order to counteract microbial colonization (Aspergillus sp., Streptomyces sp., Micrococcus sp.) on wooden artwork surfaces, Thymus vulgaris L. (Lamiaceae) essential oil (EO) and hydro-alcoholic (HA) solutions were applied in a polyphasic approach. The antimicrobial activities of EO and HA solutions were preliminarily assessed by agar disc…

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Endophytes from medicinal plants’ seeds: exploring new reservoirs of bioactive molecules

Plant-associated microorganisms have recently gained more attention for their influence on plant health and biotechnological relevance. Bioactive molecules have been already isolated from plant-associated bacteria. Seed-borne bacterial endophytes have not been much explored though. Such endophytes are particularly important, since they can influence germination and be transmitted from generation to generation. Interestingly, seed endophytes from medicinal plants could influence the production of molecules with therapeutic properties. Bacterial endophytic strains were extracted from surface-sterilized seeds of the medicinal plant Echinacea purpurea and their 16S rRNA genes were sequenced. Th…

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The seed endophytic microbiota of Citrus limon L. Burm. F

Background Plant seeds possess a complex microbiota which may play a crucial rolein a plethora of mechanisms, such as preservation, germination, seedling development, plant growth and health. In particular, seed endophytic microbiota is gaining more and more consideration due to the fact that it may be vertically transmitted to ensure individual competitive advantages. The Citrusspecies, including C.limon,areone of the most economically important evergreen fruit crops in the world. Objectives i)Identification and characterization ofmicrobial isolates from Citrus limon L. Burm. F. seed endosphere. ii) Structure of Citrus limon L. Burm. F. seed endosphytic microbiota. Methods -Surface-sterili…

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Omics approaches to elucidate the molecular physiology of lantibiotc NAI-107 production in Microbispora ATCC-PTA-5024

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The Streptomyces coelicolor Small ORF trpM Stimulates Growth and Morphological Development and Exerts Opposite Effects on Actinorhodin and Calcium-Dependent Antibiotic Production

In actinomycetes, antibiotic production is often associated with a morpho-physiological differentiation program that is regulated by complex molecular and metabolic networks. Many aspects of these regulatory circuits have been already elucidated and many others still deserve further investigations. In this regard, the possible role of many small open reading frames (smORFs) in actinomycete morpho-physiological differentiation is still elusive. In Streptomyces coelicolor, inactivation of the smORF trpM (SCO2038) – whose product modulates L-tryptophan biosynthesis – impairs production of antibiotics and morphological differentiation. Indeed, it was demonstrated that TrpM is able to interact w…

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The endophytic microbiota of Citrus limon is transmitted from seed to shoot highlighting differences of bacterial and fungal community structures

AbstractCitrus limon (L.) Burm. F. is an important evergreen fruit crop whose rhizosphere and phyllosphere microbiota  have been characterized, while seed microbiota is still unknown. Bacterial and fungal endophytes were isolated from C. limon surface-sterilized seeds. The isolated fungi—belonging to Aspergillus, Quambalaria and Bjerkandera genera—and bacteria—belonging to Staphylococcus genus—were characterized for indoleacetic acid production and phosphate solubilization. Next Generation Sequencing based approaches were then used to characterize the endophytic bacterial and fungal microbiota structures of surface-sterilized C. limon seeds and of shoots obtained under aseptic conditions fr…

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Pyrrolomycins as antimicrobial agents. Microwave-assisted organic synthesis and insights into their antimicrobial mechanism of action

Abstract New compounds able to counteract staphylococcal biofilm formation are needed. In this study we investigate the mechanism of action of pyrrolomycins, whose potential as antimicrobial agents has been demonstrated. We performed a new efficient and easy method to use microwave organic synthesis suitable for obtaining pyrrolomycins in good yields and in suitable amount for their in vitro in-depth investigation. We evaluate the inhibitory activity towards Sortase A (SrtA), a transpeptidase responsible for covalent anchoring in Gram-positive peptidoglycan of many surface proteins involved in adhesion and in biofilm formation. All compounds show a good inhibitory activity toward SrtA, havi…

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An integrated proteomic and metabolomic study to evaluate the effect of nucleus-cytoplasm interaction in a diploid citrus cybrid between sweet orange and lemon.

Key message: Our results provide a comprehensive overview how the alloplasmic condition might lead to a significant improvement in citrus plant breeding, developing varieties more adaptable to a wide range of conditions. Abstract: Citrus cybrids resulting from somatic hybridization hold great potential in plant improvement. They represent effective products resulting from the transfer of organelle-encoded traits into cultivated varieties. In these cases, the plant coordinated array of physiological, biochemical, and molecular functions remains the result of integration among different signals, which derive from the compartmentalized genomes of nucleus, plastids and mitochondria. To dissect …

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Composition and geographic variation of the bacterial microbiota associated with the coelomic fluid of the sea urchin Paracentrotus lividus

AbstractIn the present work, culture-based and culture-independent investigations were performed to determine the microbiota structure of the coelomic fluid of Mediterranean sea urchin Paracentrotus lividus individuals collected from two distinct geographical sites neighboring a high-density population bay and a nature reserve, respectively. Next Generation Sequencing analysis of 16S rRNA gene (rDNA) showed that members of the Proteobacteria, Bacteroidetes and Fusobacteria phyla, which have been previously reported to be commonly retrieved from marine invertebrates, dominate the overall population of microorganisms colonizing this liquid tissue, with minority bacterial genera exhibiting rem…

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The cypsela (achene) of Echinacea purpurea as a diffusion unit of a community of microorganisms

AbstractEchinacea purpurea is a plant cultivated worldwide for its pharmaceutical properties, mainly related to the stimulation of the immune system in the treatment of respiratory infections. The cypselas (fruits) of E. purpurea were examined in order to investigate the presence, localization and potential function(s) of endophytic microorganisms. Electron and confocal microscopy observations showed that three different components of microorganisms were associated to cypselas of E. purpurea: (i) one endocellular bacterial component in the cotyledons, enclosed within the host membrane; (ii) another more generic bacterial component adhering to the external side of the perianth; and (iii) a f…

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Proteomics to elucidate the molecular physiology of Microbispora sp. ATCC-PTA-5024, the producer of NAI-107, a very promising lantibiotic

The filamentous actinobacterium Microbispora sp. ATCC-PTA-5024 produces the lantibiotic NAI-107 (Maffioli et al., 2014), which is effective against multidrug-resistant Gram-positive pathogens (Jabés et al., 2011). In actinomycetes, the biosynthesis of antibiotics is generally elicited as a physiological response controlled by a complex regulatory network involving global regulators, playing pleiotropic roles, and pathway-specific regulators, which activate the biosynthesis of biologically active molecules (Bibb, 2005)

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