Several bacterial cultures were isolated from sponge Halichondria sp., collected from the Gujarat coast of the Indo Pacific region. These bacterial cultures were fermented in the laboratory (100 mL) and the culture filtrate was assayed for antibiotic activity against 16 strains of clinical pathogens. Bacillus sp. (SAB1), the most potent of them and antagonistic to several clinically pathogenic Gram-positive, Gram-negative bacteria and the fungus Aspergillus fumigatus was chosen for further investigation. Analysis of the nucleotide sequence of the 16S rDNA gene of Bacillus sp. SAB1 showed a strong similarity (100%) with the 16S rDNA gene of Bacillus licheniformis HNL09. The bioactive compounds produced by Bacillus licheniformis SAB1 (GenBank accession number: DQ071568) were identified as indole (1), 3-phenylpropionic acid (2) and a dimer 4,4′-oxybis[3-phenylpropionic acid] (3) on the basis of their Fourier Transform Infrared (FTIR), Nuclear Magnetic Resonance (NMR) and Electrospray Ionization Mass Spectrometer (ESI-MS) data. There is a single reference on the natural occurrence of compound 3 from the leaves of a terrestrial herb Aptenia cordifolia in the literature, so to the best of our knowledge, this is a first report of its natural occurrence from a marine source. The recovery of bacterial strains with antimicrobial activity suggests that marine-invertebrates remain a rich source for the isolation of culturable isolates capable of producing novel bioactive secondary metabolites.
Melanins are commonly produced by bacteria, fungi, plants and animals, where they play a role in many biological functions. They protect organisms against UV and ionizing radiations. Their potential applications in biotechnological industries such as cosmetics and paints, where UV protection is required, are hampered by the lack of suitable organisms or methods to produce them abundantly. We report here the production of high amounts of extracellular melanin by the fungus Gliocephalotrichum simplex in cultures supplemented with tyrosine. Their typical UV-absorbance, as well as i.r., (13)C solid-state and (1)H NMR spectra indicated that the melanin is a eumelanin, being a copolymer of dihydroxyindole carboxylic acid and dihydroxyindole, associated with some carbohydrates and proteinaceous matter. Optimal culture conditions established by a Plackett-Burman experiment, followed by a full factorial experiment based on tyrosine and peptone yielded a maximum of up to 6.6 g melanin l(-1). The high yields of extracellular melanin from G. simplex enables its use in biotechnology.
Significance and Impact of the Study: This is the first report not only on marine sponge-associated Streptomyces for anti-QS in Ps. aeruginosa but also on cinnamic acid and proline-derived linear dipeptides proline-glycine as QS inhibitors. The results reveal that marine-derived actinomycetes may not only play a role in the defensive mechanism of their host but also lead to new molecules useful in the development of novel antivirulence drugs. AbstractThe study aimed to discover quorum sensing (QS) inhibitors from marine sponge-derived actinomycetes and analyse its inhibitory activities against QSmediated virulence factors in Pseudomonas aeruginosa. Seventy-two actinomycetes isolated from marine invertebrates collected from the western coast of India were screened against the QS indicator strain Chromobacterium violaceum CV12472. Methanol extracts of 12 actinomycetes showing inhibition of violacein production were accessed for downregulation of QS-mediated virulence factors like swarming, biofilm formation, pyocyanin, rhamnolipid and LasA production in Ps. aeruginosa ATCC 27853. The isolates NIO 10068, NIO 10058 and NIO 10090 exhibited very good anti-QS activity, with NIO 10068 being the most promising one. Mass spectrometric analysis of NIO 10068 methanol extract revealed the presence of cinnamic acid and linear dipeptides proline-glycine and N-amido-a-proline in the active extract. Detailed investigation suggested that although linear dipeptide Pro-Gly is to some extent responsible for the observed biological activity, cinnamic acid seems to be the main compound responsible for it. Marine-derived actinomycetes are a potential storehouse for QS inhibitors.
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