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Biosurfactant production from marine bacteria associated with sponge Callyspongia diffusa.

ABSTRACT: Marine-derived biosurfactants have gained significant attention due to their structural and functional diversity. Biosurfactant production was performed using bacteria associated with Callyspongia diffusa, a marine sponge inhabiting the southern coast of India. A total of 101 sponge-associated bacteria were isolated on different media, of which 29 isolates showed positive result for biosurfactant production. Among the 29 positive isolates, four were selected based on highest emusification activity and were identified based on 16S rDNA sequence analysis. These isolates were identified as Bacillus subtilis MB-7, Bacillus amyloliquefaciens MB-101, Halomonas sp. MB-30 and Alcaligenes sp. MB-I9. The 16S rDNA nucleotide sequences were deposited in GenBank with accession numbers KF493730, KJ540939, KJ414418 and KJ540940, respectively. Based on the highest oil displacement activity and effective surface tension reduction potential, the isolate B. amyloliquefaciens MB-101 was selected for further optimization and structural delineation. The production of biosurfactant by the isolate was significantly enhanced up to 6.76 g/l with optimal concentration values of 2.83 % for glycerol, 2.65 % for peptone, 20.11 mM for ferrous sulfate and 74 h of incubation by employing factorial design. The structural features of the purified biosurfactant from B. amyloliquefaciens MB-101 showed similarity with lipopeptide class of biosurfactant. In conclusion, the present study emphasizes the utilization of marine sponge-associated bacteria for the production of biosurfactant that may find various applications.

SUBMITTER: Dhasayan A

PROVIDER: S-EPMC4522725 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Four sponge-associated Antarctic bacteria (i.e., Winogradskyella sp. strains CAL384 and CAL396, Colwellia sp. strain GW185, and Shewanella sp. strain CAL606) were selected for the highly mucous appearance of their colonies on agar plates. The production of extracellular polymeric substances (EPSs) was enhanced by a step-by-step approach, varying the carbon source, substrate and NaCl concentrations, temperature, and pH. The EPSs produced under optimal conditions were chemically characterized, resulting in a moderate carbohydrate content (range, 15 to 28%) and the presence of proteins (range, 3 to 24%) and uronic acids (range, 3.2 to 11.9%). Chemical hydrolysis of the carbohydrate portion revealed galactose, glucose, galactosamine, and mannose as the principal constituents. The potential biotechnological applications of the EPSs were also investigated. The high protein content in the EPSs from Winogradskyella sp. CAL384 was probably responsible for the excellent emulsifying activity toward tested hydrocarbons, with a stable emulsification index (E24) higher than those recorded for synthetic surfactants. All the EPSs tested in this work improved the freeze-thaw survival ratio of the isolates, suggesting that they may be exploited as cryoprotection agents. The addition of a sugar in the culture medium, by stimulating EPS production, also allowed isolates to grow in the presence of higher concentrations of mercury and cadmium. This finding was probably dependent on the presence of uronic acids and sulfate groups, which can act as ligands for cations, in the extracted EPSs.IMPORTANCE To date, biological matrices have never been employed for the investigation of EPS production by Antarctic psychrotolerant marine bacteria. The biotechnological potential of extracellular polymeric substances produced by Antarctic bacteria is very broad and comprises many advantages, due to their biodegradability, high selectivity, and specific action compared to synthetic molecules. Here, several interesting EPS properties have been highlighted, such as emulsifying activity, cryoprotection, biofilm formation, and heavy metal chelation, suggesting their potential applications in cosmetic, environmental, and food biotechnological fields as valid alternatives to the commercial polymers currently used.

Project description:Three methods were examined to cultivate bacteria associated with the marine sponge Haliclona (gellius) sp.: agar plate cultures, liquid cultures, and floating filter cultures. A variety of oligotrophic media were employed, including media with aqueous and organic sponge extracts, bacterial signal molecules, and siderophores. More than 3,900 isolates were analyzed, and 205 operational taxonomic units (OTUs) were identified. Media containing low concentrations of mucin or a mixture of peptone and starch were most successful for the isolation of diversity, while the commonly used marine broth did not result in a high diversity among isolates. The addition of antibiotics generally led to a reduced diversity on plates but yielded different bacteria than other media. In addition, diversity patterns of isolates from agar plates, liquid cultures, and floating filters were significantly different. Almost 89% of all isolates were Alphaproteobacteria; however, members of phyla that are less commonly encountered in cultivation studies, such as Planctomycetes, Verrucomicrobia, and Deltaproteobacteria, were isolated as well. The sponge-associated bacteria were categorized into three different groups. The first group represented OTUs that were also obtained in a clone library from previously analyzed sponge tissue (group 1). Furthermore, we distinguished OTUs that were obtained from sponge tissue (in a previous study) but not from sponge isolates (group 2), and there were also OTUs that were not obtained from sponge tissue but were obtained from sponge isolates (group 3). The 17 OTUs categorized into group 1 represented 10 to 14% of all bacterial OTUs that were present in a large clone library previously generated from Haliclona (gellius) sp. sponge tissue, which is higher than previously reported cultivability scores for sponge-associated bacteria. Six of these 17 OTUs were not obtained from agar plates, which underlines that the use of multiple cultivation methods is worthwhile to increase the diversity of the cultivable microorganisms from sponges.

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Biosurfactant production from marine bacteria associated with sponge Callyspongia diffusa.

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