Search

Seaweed-associated bacterial symbionts are sources of potential pharmacological properties. The present study resulted in the culture-dependent isolation of bioactive heterotrophs belonging to the bacterial phylum Firmicutes, which were dominated more than 30% of the 127 cultivable isolates, amongst which 23 of them showed potential antimicrobial activities against a wide range of pathogens. The symbionts isolated from the seaweed Sargassum wightii showed significant bioactivity. Those were characterized as Bacillus safensis MTCC13040, B. valismortis MTCC13041, B. velezensis MTCC13044, B. methylotrophicus MTCC13042, Oceanobacillus profundus MTCC13045, B. tequilensis MTCC13043, and B. altitudinis MTCC13046. The organic extracts of the studied isolates showed potential antimicrobial properties against methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococci (minimum inhibitory concentration 6·25-12·5 μg ml -1 ). The organic extract of B. altitudinis MTCC13046 displayed significantly greater radical quenching ability (IC 90 133 μg ml -1 , P < 0·05) other than attenuating hydroxymethyl glutaryl coenzyme A reductase (IC 90 10·21 μg ml -1 , P < 0·05) and angiotensin-converting enzyme-1 (IC 90 498 μg ml -1 , P < 0·05) relative to other studied heterotrophs. The organic extract of B. tequilensis MTCC13043 displayed significantly greater attenuation potential against pro-inflammatory 5-lipooxygenase (IC 90 5·94 μg ml -1 , P < 0·05) and dipeptidyl peptidase-4 (IC 90 271 μg ml -1 , P < 0·05). The seaweed-associated B. altitudinis MTCC13046 and B. tequilensis MTCC13043 could be used to develop promising pharmacological leads., (© 2022 Society for Applied Microbiology.)

The inevitability to develop novel antimicrobial agents has considerably increased because of mounting alarms concerning multidrug-resistant microbial strains. The present study evaluated an antibacterial and wound healing topical formulation prepared with the ethyl acetate extract of marine symbiotic Bacillus amyloliquefaciens MTCC 12716 as the basic ingredient and the grafted macroalgal polysaccharide as the gel base with an appropriate proportion of natural stabilizing agents. The formulation exhibited potent antibacterial activities against clinical isolates of Staphylococcus aureus (18 mm inhibition zone) and Pseudomonas aeruginosa (19 mm) causing infection when compared with commercially available antimicrobial cream clindamycin. The in-vitro results indicated that the organic extract of B. amyloliquefaciens MTCC 12716 at its MIC and the formulation sealed the wound by 78 and 94%, respectively, at 48 h in the scratch-induced L929 cells, compared to 84% exhibited by clindamycin. The topical formulation of marine symbiotic Bacillus induced greater than 80% viability of the normal fibroblasts compared to 78% exhibited by clindamycin, when administered at a dose of 25 μg mL -1 . The studied antibacterial formulation could accelerate the wound healing by prompting the migration of fibroblasts towards the artificially created wound resulting in rapid wound closure, and at an even higher concentration of formulation, it displayed no cytotoxicity on L929 cells. The stability studies showed that the formulation maintained its physicochemical characteristics and minimal growth (<10 cfu g -1 ) of bacteria on the plates throughout the time period of 18 months at 30 °C and 65% relative humidity. This study has established the antibacterial and wound healing potential of a topical formulation of marine symbiotic B. amyloliquefaciens., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Emergence of multidrug-resistant microorganisms and requirements for novel antimicrobial compounds necessitate exploring newer habitats to develop potential bioactive leads. Culture-contingent analysis of heterotrophic bacterial flora from the seaweeds led to the isolation of bioactive strains possessing potential antibacterial properties against wide-ranging clinical pathogens viz., methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus faecalis (VREfs). Seven of the most active strains belonging to the phylum Firmicutes isolated from a brown seaweed (Phaeophyceae) Sargassum wightii exhibited spot-over-lawn assay guided inhibition zone of larger than 30 mm. Integrated phenotypic and genotypic studies have led to the characterization of the seaweed-associated bacteria particularly belonging to the phylum Firmicutes. The organic extracts of the studied bacteria exhibited promising antibacterial properties against MRSA and VREfs with minimum inhibitory concentration ranging between 6.25 and 12.50 μg/mL. Time-kill kinetic profiles of those bacteria displayed rapid bactericidal activity against both E. coli and MRSA, showing a ≥ 3log 10 reduction in viable cell count than the initial. Among the studied bioactive Bacillus spp, B. tequilensis MTCC13043 and B. altiitudinis MTCC13046 were found to possess functional polyketide synthase (pks) gene (MW027664 and MW027660) that could be amplified. The outcome of amplified genes encrypting for polyketide synthase in conjunction with antibacterial activities unveiled the broad-spectrum antimicrobial activities of the marine heterotrophic Firmicutes, which could be further used against the emergent problem of antibiotic-resistant bacterial pathogens., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Appearance of drug-resistant microorganisms prompted researchers to unravel new environments for development of novel antimicrobial agents. Culture-supported analysis of heterotrophic bacteria associated with seaweeds yielded 152 strains, in that larger share of the isolates was embodied by Bacillus atrophaeus SHB2097 (54%), B. velezensis SHB2098 (24%), B. subtilis SHB2099 (12%), and B. amyloliquefaciens SHB20910 (10%). One of the most active strains characterized as B. atrophaeus SHB2097 (MW821482) with an inhibition zone more than 30 mm on spot-over-lawn experiment, was isolated from a seaweed Sargassum wightii, was selected for bioprospecting studies. Significant antibacterial potential was displayed by bacterial organic extract against vancomycin-resistant Enterococcus faecalis, Pseudomonas aeruginosa, methicillin-resistant Staphylococcus aureus, and Klebsiella pneumonia with minimum inhibitory concentration 6.25 µg/mL and comparable to the antibiotics ampicillin and chloramphenicol. The genes of type 1 pks (MZ222383, 700 bp) and hybrid nrps/pks (MZ222389, 1000-1400 bp) of B. atrophaeus MW821482 could be amplified. The bacterium displayed susceptibility to the commercially available antibiotic agents, and was negative for the pore-forming non-hemolytic hemolysin BL (hbl) and enterotoxin (nhe) genes, and therefore, was not pathogenic. The bacterium was found to possess genes (1000-1400 bp) involved in the biosynthesis of siderophore-class of compounds (MZ222387 and MZ222388) that showed 99% of similarity in BLAST search, and showed production of siderophore. Noteworthy antibacterial activities against clinically important pathogenic bacteria in conjunction with occurrence of genes coding for antimicrobial metabolites inferred that the marine heterotrophic bacterium B. atrophaeus SHB2097 could be used for the development of antibacterial agents against the emerging antibiotic resistance., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Increased prevalence of microbial resistance and development of drug-resistant pathogens have triggered an urge among researchers to discover potential antimicrobial compounds, particularly from the marine habitat. The present study highlights the cultivable diversity and bioactivities of heterotrophic bacteria associated with marine macroalgae of southeast Indian coastal region. Culture-dependent isolation method resulted in 40 isolates, in which greater part of the isolates represented Gammaproteobacteria (62%) followed by that comprised of the phylum Firmicutes. One of the most active strains isolated from a macroalga, Laurencia papillosa, was characterized based on the integrated phenotypic and genotypic analysis as Bacillus velezensis MBTDLP1 MTCC 13048, with an inhibition zone of about 35 mm against methicillin-resistant Staphylococcus aureus (MRSA), was selected for bioprospecting studies. Type-I pks gene (MT394492) of 700 bp could be amplified from the heterotrophic B. velezensis. The bacterium exhibited siderophore production and possessed genes implicated in the biosynthesis of siderophore type of metabolites exhibiting 99% similarity with other GenBank sequences in BLAST search. B. velezensis exhibited promising anti-infective properties against methicillin-resistant Staphylococcus aureus (minimum inhibitory concentration 15 µg/mL), and the activities were positively correlated (r 2  > 0.9) with iron-chelating activities. Chemical investigation of the organic extract of B. velezensis MBTDLP1 characterized a macrocyclic polyketide exhibiting prospective antibacterial potential against methicillin-resistant S. aureus (MIC 0.38 µg/mL), than that exhibited by positive control chloramphenicol (6.25 µg/mL). Significant antibacterial activity against drug-resistant bacteria combined with the presence of genes coding for bioactive secondary metabolites revealed that this marine symbiotic bacterium could be used against emerging antibiotic resistance., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The rise in antibiotic-resistant bacterial strains prompting nosocomial infections drives the search for new bioactive substances of promising antibacterial properties. The surfaces of seaweeds are rich in heterotrophic bacteria with prospective antimicrobial substances. This study aimed to isolate antibacterial leads from a seaweed-associated bacterium. Heterotrophic Bacillus amyloliquefaciens MTCC 12716 associated with the seaweed Hypnea valentiae, was isolated and screened for antimicrobial properties against drug-resistant pathogens. The bacterial crude extract was purified and three novel amicoumacin-class of isocoumarin analogues, 11'-butyl acetate amicoumacin C (amylomacin A), 4'-hydroxy-11'-methoxyethyl carboxylate amicoumacin C (amylomacin B) and 11'-butyl amicoumacin C (amylomacin C) were isolated to homogeneity. The studied amylomacins possessed potential activities against Pseudomonas aeruginosa, vancomycin-resistant Enterococcus faecalis, Klebsiella pneumoniae, methicillin-resistant Staphylococcus aureus, and Shigella flexneri with a range of minimum inhibitory concentration values from 0.78 to 3.12 µg/mL, although standard antibiotics ampicillin and chloramphenicol were active at 6.25-25 µg/mL. Noticeably, the amylomacin compound encompassing 4'-hydroxy-11'-methoxyethyl carboxylate amicoumacin C functionality (amylomacin B), displayed considerably greater antagonistic activities against methicillin-resistant S. aureus, vancomycin-resistant E. faecalis, Vibrio parahaemolyticus, Escherichia coli, and K. pneumoniae (minimum inhibitory concentration 0.78 μg/mL) compared to the positive controls and other amylomacin analogues. Antimicrobial properties of the amylomacins, coupled with the presence of polyketide synthase-I/non-ribosomal peptide synthetase hybrid gene attributed the bacterium as a promising source of antimicrobial compounds with pharmaceutical and biotechnological applications., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

Ever since the development of the first antibiotic compound with anticancer potential, researchers focused on isolation and characterization of prospective microbial natural products with potential anti-infective and anticancer activities. The present work describes the production of bioactive metabolites by heterotrophic bacteria associated with intertidal seaweeds with potential anti-infective and anticancer activities. The bacteria were isolated in a culture-dependent method and were identified as Shewanella algae MTCC 12715 (KX272635) and Bacillus amyloliquefaciens MTCC 12716 (KX272634) based on combined phenotypic and genotypic methods. Further, the bacteria were screened for their ability to inhibit drug-resistant infectious pathogens and prevent cell proliferation of human liver carcinoma (HepG2) and breast cancer (MCF7) cell lines, without affecting the normal cells. Significant anti-infective activity was observed with bacterial cells and their organic extracts against broad-spectrum multidrug-resistant pathogens, such as vancomycin-resistant Enterococcus faecalis, methicillin-resistant Staphylococcus aureus, Klebsiella pneumonia and Pseudomonas aeruginosa with minimum inhibitory concentration ≤ 3.0 µg mL -1 as compared to the antibiotic agents' chloramphenicol and ampicillin, which were active at ≥ 6.25 mg mL -1 . The extracts also exhibited anticancer activity in a dose-responsive pattern against HepG2 (with IC 50 , half maximal inhibitory concentration ~ 78-83 µg mL -1 ) and MCF7 (IC 50  ~ 45-48 µg mL -1 ) on tetrazolium bromide screening assay with lesser cytotoxic effects on normal fibroblast (L929) cell lines (IC 50  > 100 µg mL -1 ). The results revealed that seaweed-associated heterotrophic bacteria could occupy a predominant role for a paradigm shift towards the development of prospective anti-infective and anticancer agents.

Three homologous oxygenated elansolid-type of polyketide spanned macrolides were isolated from a heterotrophic marine bacterium, Bacillus amyloliquefaciens MTCC 12716, associated with an intertidal red alga Hypnea valentiae. The complete genome of the bacterium was sequenced and all detectable natural product gene clusters were analysed. The B. amyloliquefaciens MTCC 12716 genome features polyketide synthase (pks) systems of every known formally classified family, nonribosomal peptide synthetases and hybrid clusters. Comprehensive spectroscopic studies revealed the compounds to possess isobenzofuranyl benzoate and 1H-furopyrano[2,3-c]oxacyclononadecine-6-carboxylate moieties. The identified compounds displayed broad-spectrum bactericidal activity against methicillin-resistant Staphylococcus aureus, vancomycin-resistant Enterococcus faecalis, and drug-resistant strains of Pseudomonas aeruginosa and Klebsiella pneumoniae with minimum inhibitory concentrations (MICs) of ≤1.0 µg/mL, whereas the standard antibiotics ampicillin and chloramphenicol were active only at concentrations of ≥6.25 µg/mL. The plausible mechanism of elansolid-type macrolide biosynthesis by trans-AT polyketide synthases through the pks starter unit para-hydroxybenzoic acid was hypothesised, and the structures were correlated with the gene organisation, with the predicted gene cluster comprising 16 genes (~81 kb in size). The best binding poses for each compound with the peptide deformylase (PDF) protein of S. aureus revealed docking scores (>11.30 kcal/mol) greater than actinonin (6.96 kcal/mol), a natural PDF inhibitor. The higher electronic values along with optimum lipophilic parameters support the potential anti-infective properties of the studied macrolides. These antibacterial elansolid-type of polyketide spanned macrolides in marine symbiotic B. amyloliquefaciens could be potential leads for biotechnological and pharmaceutical applications against emerging multidrug-resistant pathogens., (Copyright © 2020 Elsevier B.V. and International Society of Chemotherapy. All rights reserved.)

Aims: Microbiological, biotechnological and chemical characterization of a previously undescribed aryl-crowned polyketide from Bacillus subtilis MTCC 10403 isolated from brown seaweed Anthophycus longifolius with activity against opportunistic Gram-negative food-borne pathogenic bacterial strains., Methods and Results: A culture-dependent method was used to isolate heterotrophic B. subtilis associated with A. longifolius and assessed for its antimicrobial properties. Minimum inhibitory concentration (MIC) of the title compound against the test pathogens was analysed by microtube dilution coupled with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide-based colorimetric endpoint detection. Bacillus subtilis MTCC 10403 was found to be antagonistic against Gram-negative food-borne pathogenic Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Salmonella enterica serotype Typhi, Aeromonas hydrophila and Vibrio sp. (diameter of zone of growth inhibition 13-22 mm). Bacillus subtilis was assessed for the presence of secondary metabolite coding polyketide synthase (pks) gene (KC589397, 700-bp gene product) and carboxylate siderophore framework in the aryl-crowned polyketide designated as 7-O-6'-(2″-acetylphenyl)-5'-hydroxyhexanoate-macrolactin by exhaustive spectroscopic techniques. The MIC assay showed that the reference antibiotics tetracycline and ampicillin were active at 25 μg ml -1 against the test pathogens, whereas the newly isolated polyketide displayed anti-infective properties against E. coli, A. hydrophilla, P. aeruginosa and Vibrio sp. at a lower concentration (MIC <13 μg ml -1 ). The MIC of the aryl macrolactin against K. pneumoniae was comparable with that of the referral antibiotics (~25 μg ml -1 ). The mode of antimicrobial action of acryl-crowned macrolactin was found to be iron chelating similar to siderophores. Putative biosynthetic pathway of the pks gene product further validated its molecular attributions., Conclusions: This study recognized new variant of antimicrobial aryl-crowned polyketide bearing methyl 6'-(2″-acetylphenyl)-5'-hydroxyhexanoate moiety at the C-7 position of the macrolactin system from A. longifolius-associated bacterium B. subtilis., Significance and Impact of the Study: This study revealed seaweed-associated micro-organisms as promising biological strata to produce new-generation anti-infective agents., (© 2017 The Society for Applied Microbiology.)

Brown seaweed Anthophycus longifolius (Turner) Kützing (family Sargassaceae) associated heterotrophic bacterium Bacillus subtilis MTCC 10403 was found to be a potent isolate with broad range of antibacterial activity against important perceptive food pathogens Vibrio parahaemolyticus, V. vulnificus, and Aeromonas hydrophila. This bacterium was positive for polyketide synthetase gene (KC589397), and therefore, was selected to bioprospect specialized metabolites bearing polyketide backbone. Bioactivity-guided chromatographic fractionation of the ethyl acetate extract of the seaweed-associated bacterium segregated four homologous polyketide furanoterpenoids with potential antibacterial activities against clinically important pathogens. The minimum inhibitory concentration (MIC) assay showed that the referral antibiotics tetracycline and ampicillin were active at 25 μg/mL against the test pathogens, whereas the previously undescribed (4E)-methyl 13-((16-(furan-2-yl) ethyl)-octahydro-7-hydroxy-4-((E)-23-methylbut-21-enyl)-2H-chromen-6-yl)-4-methylpent-4-enoate (compound 1) and methyl 3-(hexahydro-9-((E)-3-methylpent-1-enyl)-4H-furo[3,2-g]isochromen-6-yl) propanoate (compound 3) displayed antibacterial activities against the test pathogens at a lesser concentration (MIC < 7 μg/mL). The title compounds were characterized by comprehensive nuclear magnetic resonance and mass spectroscopic experiments. Polyketide synthase catalyzed putative biosynthetic mechanism additionally corroborated the structural ascriptions of the hitherto undescribed furanoterpenoids from seaweed-associated bacterial symbiont. The electronic and hydrophobic parameters appeared to hold a conspicuous part in directing the antibacterial properties of the compounds. Seaweed-associated B. subtilis MTCC 10403 demonstrated to represent a potential source of antimicrobial polyketides for pharmaceutical applications., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

In this study, 234 bacterial strains were isolated from 7 seaweed species in the Gulf of Mannar on the southeast coast of India. The strains having consistent antimicrobial activity were chosen for further studies, and this constituted about 9.8% of the active strains isolated. Phylogenetic analysis using 16S rDNA sequencing with the help of classical biochemical identification indicated the existence of 2 major phyla, Firmicutes and Proteobacteria. Antimicrobial activity analysis combined with the results of amplifying genes encoding for polyketide synthetase and nonribosomal peptide synthetase showed that seaweed-associated bacteria had broad-spectrum antimicrobial activity. These epibionts might be beneficial to seaweeds by limiting or preventing the development of competing or fouling bacteria. Phylogenetic analysis of ketosynthase (KS) regions with respect to the diverse range of KS domains showed that the KS domains from the candidate isolates were of Type I. The bacterial cultures retained their antimicrobial activities after plasmid curing, which further suggested that the antimicrobial activity of these isolates was not encoded by plasmid, and the genes encoding the antimicrobial product might be present within the genome. Seaweed-associated bacteria with potential antimicrobial activity suggested that the seaweed species are an ideal ecological niche harboring specific bacterial diversity representing a largely underexplored source of antimicrobial secondary metabolites.