Cytotoxicity of di-rhamnolipids produced by Pseudomonas aeruginosa RA5 against human cancerous cell lines.

Rhamnolipid biosurfactant produced by Pseudomonas aeruginosa, possesses non-toxicity, environmental compatibility, a wide range of pH (4-8), temperature (4-100 °C), and salinity (1-10%) stability. The application of RLs is worldwide accepted in the pharmaceutical, medicinal, and food industries. It has been used for cytotoxicity efficacy analysis with a limited number of cancerous cell lines. To widen the scope of rhamnolipid application as an anticancer agent, we have studied Di-RLs homolog, 'Rha-Rha-C10-C10' produced by Pseudomonas aeruginosa RA5 against human cancerous cell lines including breast cancer (MCF-7), leukemia (K-562), cervical cancer (HeLa), Lung cancer (HOP-62), and colon cancer (HT-29) in a dose-dependent way. It was purified with silica gel chromatography followed by TLC and mass spectroscopy prior to cytotoxicity analysis. With a tensiometer, critical micelle concentration of Di-RLs was estimated to be 33.92 ± 2 mN/m at 0.2%. Cytotoxicity analysis of Di-RLs on K-562 cell line demonstrated inhibition with GI50 and TGI at < 10 µg/mL and 66.6 µg/mL, after 48 h of application. The morphology of human cancerous cell lines was observed under a laser confocal microscope with the SRB staining method. Further research is recommended to comprehend the Di-RLs as a potential anti-cancer agent.

Development of miniaturized agar based assays in 96-well microplates applicable to high-throughput screening of industrially valuable microorganisms.

High-throughput screening (HTS) is a present-day approach for assaying thousands of cultures in parallel. This miniaturization allows rapid screening of large number of microorganims capable of producing bio-based materials thereby meeting the demands of the ever evolving food, pharmaceutical and cosmetic industry. In this study, agar-based assays for phosphate solubilization, cellulose degradation and lactic acid production were developed in 96-well microplates using Biomek FXP Automated Liquid Handling system. Techno-economic analysis from this study reveals the lower overall cost per assay using HTS as compared to conventional Petri plate assays. Though automated liquid handling workstations have been used to perform liquid-based assays, there are very few studies which report their use for agar-based microplate assays. These findings thus corroborate the establishment of rapid and efficient miniaturized, qualitative agar-based screening methods for identifying microorganisms with potential for commercial application.

Genomic insights into biocontrol potential of edible seaweed-associated Bacillus velezensis MTCC 10456 from Gulf of Mannar.

Bacillus velezensis MTCC 10456, is a marine mesophilic heterotrophic bacterium, isolated from edible red seaweed, Laurenciae papillosa, with a potential for plant growth promotion and biocontrol activity. We report the genome sequence analysis of strain MTCC 10456, which has a genome size of 4.19 Mb with an average G + C content of 45.9%, 4077 coding sequences and 94 RNAs. Comparative genome analysis of MTCC 10456 with 76 other land plant-based strains with complete information on the source of their isolation, was carried out. This study provided evidence that multiple gene clusters that contributed to the seaweed colonization, growth promotion, immunity and hyperosmotic stress tolerance are conserved in different B. velezensis strains. A unique methyltransferase gene that can cause global alterations in DNA methylation patterns affecting gene expression and regulation of transcription in MTCC 10456 genome was identified. The genome provides insights into multiple gene clusters encoding antagonistic metabolites, such as non-ribosomal peptide synthetases (NRPSs) and polyketide synthetases (PKSs), unidentified metabolites and genes for antimicrobial resistance. Overall, findings from this study indicate the application of B. velezensis MTCC 10456 in the aquaculture industry as a biocontrol agent and may also contribute in understanding the ecological adaptations of plant-associated B. velezensis strains in different habitats.

Antagonistic Activity of Antimicrobial Metabolites Produced from Seaweed-Associated Bacillus amyloliquefaciens MTCC 10456 Against Malassezia spp.

Members of the genus Malassezia are known to be opportunistic pathogens responsible for causing skin disorders such as seborrheic dermatitis or dandruff, pityriasis versicolor, folliculitis, atopic dermatitis, and psoriasis. Due to the side effects caused by prolonged use of current topical antifungal agents, development of an alternative treatment is necessary. Fermentative production of antimicrobial metabolites from Bacillus amyloliquefaciens MTCC 10456 was carried out, and their antagonistic activity against Malassezia furfur and Malassezia globosa was evaluated. The antifungal metabolites were isolated by acid precipitation, and bioassay-guided simultaneous separation of the antimicrobial compounds was done by reversed-phase high-performance liquid chromatography (RP-HPLC). The fraction which demonstrated antifungal activity consisted of bacilysin, homologues of bacillomycin D, and members of the macrolactin family. The presence of bacilysin was detected using specific inhibitor assays and homologues of bacillomycin D, and macrolactins were identified using liquid chromatography/high-resolution electrospray ionization-mass spectrometry (LC/HRESI-MS/MS) analysis. Synergism among the identified compounds was observed which enhanced the antagonistic activity against Malassezia spp. To our knowledge, this is the first study to report the co-production and separation of members of macrolactin antibiotics, lipopeptides such as bacillomycin D and dipeptide antibiotic bacilysin, by any Bacillus strain from marine environment. Activity of individual compounds against Malassezia has been reported which may facilitate their application in the field of dermatology and in cosmetic products.

Antibacterial Co(II), Ni(II), Cu(II) and Zn(II) complexes of Schiff bases derived from fluorobenzaldehyde and triazoles.

Antibacterial Schiff bases derived from 1,2,4-triazoles as well as their metal complexes incorporating cobalt(II), nickel(II), copper(II) and zinc(II) have been synthesized and characterized. Physico-chemical studies suggest that an octahedral geometry for the cobalt(II), nickel(II) and zinc(II)and square-planer geometry for the copper(II) complexes. These complexes have been screened for antibacterial activity against three Gram-positive (Staphylococcus aureus, Staphylococcus epidermidis and Bacillus subtilis) and two Gram-negative (Salmonella typhi and Pseudomonas aeruginosa) bacterial strains, and results compared with the activity of the free ligands. The metal complexes were found to be more potent against one or more bacterial strains than the free ligands.

Some bivalent metal complexes of Schiff bases containing N and S donor atoms.

Schiff bases have been synthesized by the reaction of p-nitrobenzaldehyde, o-nitrobenzaldehyde and p-toluyaldehyde with 4-amino-5-mercapto-1,2,4-triazole. The ligands react with Co(II), Ni(II) and Zn(II) metals to yield (1:1) and (1:2) [metal:ligand] complexes. Elemental analyses, IR, 1H NMR, electronic spectral data, magnetic susceptibility measurements, molar conductivity measurements and thermal studies have investigated the structure of the ligands and their metal complexes. The electronic spectral data suggests octahedral geometry for Co(II), Ni(II) and Zn(II). The antibacterial activities of the ligands and their metal complexes have been screened in vitro against three Gram-positive (Staphylococcus aureus, Staphylococcus epidermidis and Bacillus subtilis) and two Gram-negative (Salmonella typhi and Pseudomonas aeruginosa) organisms. The coordination of the metal ion had a pronounced effect on the microbial activities of the ligands and the metal complexes have higher antimicrobial effect than the free ligands.

Hypervalent iodine mediated synthesis of 1-aryl/hetryl-1,2,4-triazolo[4,3-a] pyridines and 1-aryl/hetryl 5-methyl-1,2,4-triazolo[4,3-a]quinolines as antibacterial agents.

Oxidation of 2-pyridyl and 2-quinylhydrazones with iodobenzene diacetate (IBD) in dichloromethane yield 1-aryl/hetryl-1,2,4-trizolo-[4,3-a] pyridines (3a-f) and 1-aryl/hetryl-5-methyl-1,2,4-triazolo[4,3-a] quinolines (6a-f). Seven compounds were tested in vitro for their antibacterial activity. 1-(5'-Nitro-2-furyl)-5-methyl-1,2,4-triazolo[4,3-a]quinoline (6e) was associated with substantially higher antibacterial activity than some commercial antibiotics against Salmonella typhi at MIC i.e. 10 microg mL(-1).