Single step concomitant concentration, purification and characterization of two families of lipopeptides of marine origin.

The extracellular biosurfactant product secreted by a marine bacterium was concentrated and purified directly from the fermentation broth in a single step by ultrafiltration (UF) employing YM 30 kDa (UF-I) and Omega 10 kDa (UF-II) polyethersulfone membranes. The optimum operating pressure required for both membranes, UF-I and UF-II, were found to be 30 and 35 psi, respectively. The biosurfactant from the fermentation broth was recovered in higher amounts using UF-II (89%) than using UF-I (73%). An analysis of the critical micelle concentrations (CMC) of the recovered lipopeptides showed a lower CMC value of 15 mg L⁻¹ for the UF-II product, indicating higher degree of purity (83%) when compared to that of the UF-I product (78%). The ultrafiltered products were characterized using Fourier transformed infrared spectroscopy and matrix-assisted laser desorption ionization time of flight mass spectral analysis, which demonstrated the presence of two families of lipopeptides.

Biosurfactant production and growth kinetics of bacteria in a designer marine medium: improved physiochemical properties.

A combinatorial screening strategy was adopted for the development of a suitable medium for enhanced biosurfactant production by a marine strain. As a result, a modified marine medium (MMM) was developed, which contained urea and strontium chloride besides other salts important for the growth of marine bacteria. This medium supported growth, evident from a higher maximum growth rate value of 0.42 h(-1) and an enhanced biosurfactant production of 2.58 g/L. The critical micelle concentration (CMC) was determined for the biosurfactants obtained from all tested media combinations. The biosurfactant produced with this medium was stable at high temperature (100 °C), a wide range of pH (5-11) and salt concentration of 5-35%. The emulsifying activity and stability of the biosurfactant obtained using MMM was better than the biosurfactant obtained using conventional media. This biosurfactant with improved physiochemical properties is suitable for a wide range of applications in industry and for marine environmental cleaning.

Microbial surfactants of marine origin: potentials and prospects.

Marine environment occupies the vast majority of the earth's surface and is a rich source of highly potent and active compounds. In recent years, microbial surfactants and emulsifiers have been reported from marine microflora. Surfactant and emulsifier molecules having diverse chemical nature such as exopolysaccharides, carbohydrate-lipid-protein complexes or glycolipopeptide, glycolipids, lipopeptides, phospholipids and ornithine lipids have been reported from various marine bacteria. These surface-active agents have been found to possess good emulsification and stabilization potentials for various lipophilic compounds such as aliphatic, aromatic and polyaromatic hydrocarbons and their uptake and degradation by the microorganisms. Few biosurfactant types such as glycolipids and lipopeptides have also been found to possess valuable biological activities. Surface-active agents from marine environments thus have tremendous potential to be used in industrial processes, for environmental remediation and as drugs.

Artificial neural network modeling and genetic algorithm based medium optimization for the improved production of marine biosurfactant.

A nonlinear model describing the relationship between the biosurfactant concentration as a process output and the critical medium components as the independent variables was developed by artificial neural network modeling. The model was optimized for the maximum biosurfactant production by using genetic algorithm. Based on a single-factor-at-a-time optimization strategy, the critical medium components were found to be glucose, urea, SrCl(2) and MgSO(4). The experimental results obtained from a statistical experimental design were used for the modeling and optimization by linking an artificial neural network (ANN) model with genetic algorithm (GA) in MATLAB. Using the optimized concentration of critical elements, the biosurfactant yield showed close agreement with the model prediction. An enhancement in biosurfactant production by approximately 70% was achieved by this optimization procedure.

High-performance liquid chromatography purification of biosurfactant isoforms produced by a marine bacterium.

A marine Bacillus strain produced biosurfactant during its growth in a defined glucose-containing medium. An efficient method for separation and purification of biosurfactant isoforms was developed and optimized in high-performance liquid chromatography (HPLC) by manipulating solvent gradient program and flow rates. Starting with an initial run time of 60 min, the final optimized method had a significantly reduced run time of 20 min. By using this method, all the surface-active isoforms (fractions A-D) were eluted within 12 min of elution with much shortened retention time of each component. The purity levels of the isoforms were enhanced using the optimized method as evident from their lower CMC values. Among the four surface-active fractions, antimicrobial action was solely displayed by HPLC fraction A. FTIR analysis revealed all the HPLC fractions to be lipopeptide in nature and MALDI-ToF mass spectral analysis showed that these belonged to the fengycin family containing C(15), C(16), and C(17) fengycins.

Antimicrobial biosurfactants from marine Bacillus circulans: extracellular synthesis and purification.

AIMS: To purify the biosurfactant produced by a marine Bacillus circulans strain and evaluate the improvement in surface and antimicrobial activities. METHODS AND RESULTS: The study of biosurfactant production by B. circulans was carried out in glucose mineral salts (GMS) medium using high performance thin layer chromatography (HPTLC) for quantitative estimation. The biosurfactant production by this strain was found to be growth-associated showing maximum biosurfactant accumulation at 26 h of fermentation. The crude biosurfactants were purified using gel filtration chromatography with Sephadex G-50 matrix. The purification attained by employing this technique was evident from UV-visible spectroscopy and TLC analysis of crude and purified biosurfactants. The purified biosurfactants showed an increase in surface activity and a decrease in critical micelle concentration values. The antimicrobial action of the biosurfactants was also enhanced after purification. CONCLUSIONS: The marine B. circulans used in this study produced biosurfactants in a growth-associated manner. High degree of purification could be obtained by using gel filtration chromatography. The purified biosurfactants showed enhanced surface and antimicrobial activities. SIGNIFICANCE AND IMPACT OF THE STUDY: The antimicrobial biosurfactant produced by B. circulans could be effectively purified using gel filtration and can serve as new potential drugs in antimicrobial chemotherapy.