Production and Properties of a Surface-Active Lipopeptide Produced by a New Marine Brevibacterium luteolum Strain

Microbial-derived surfactants are molecules of great interest due to their environmentally friendly nature and low toxicity; however, their production cost is not competitive when compared to synthetics. Marine microorganisms are exposed to extremes of pressure, temperature, and salinity; hence, they can produce stable compounds under such conditions that are useful for industrial applications. A screening program to select marine bacteria able to produce biosurfactant using low-cost substrates (mineral oil, sucrose, soybean oil, and glycerol) was conducted. The selected bacterial strain showed potential to synthesize biosurfactants using mineral oil as carbon source and was identified as Brevibacterium luteolum. The surface-active compound reduced the surface tension of water to 27 mN m^?1 and the interfacial tension (water/hexadecane) to 0.84 mN m^?1 and showed a critical micelle concentration of 40 mg L^?1. The biosurfactant was stable over a range of temperature, pH, and salt concentration and the emulsification index (E₂₄) with different hydrocarbons ranging from 60 to 79 %. Structural characterization revealed that the biosurfactant has a lipopeptide nature. Sand washing removed 83 % of crude oil demonstrating the potential of the biosurfactants (BS) for bioremediation purposes. The new marine B. luteolum strain showed potential to produce high surface-active and stable molecule using a low-cost substrate.     

Application of Biosurfactant from Sphingobacterium spiritivorum AS43 in the Biodegradation of Used Lubricating Oil

This study aimed at investigating the application of biosurfactant from Sphingobacterium spiritivorum AS43 using molasses as a substrate and fertilizer to enhance the biodegradation of used lubricating oil (ULO). The cell surface hydrophobicity of bacteria, the emulsification activity, and the biodegradation efficiency of ULO were measured. The bacterial adhesion in the hydrocarbon test was used to denote the cell surface hydrophobicity of the used bacterial species. The results indicate a strong correlation between cell surface hydrophobicity, emulsification activity, and the degree of ULO biodegradation. The maximum degradation of ULO (62 %) was observed when either 1.5 % (w/v) of biosurfactant or fertilizer was added. The results also revealed that biosurfactants alone are capable of promoting biodegradation to a large extent without added fertilizer. The data indicate the potential for biosurfactant production by using low-cost substrate for application in the bioremediation of soils contaminated with petroleum hydrocarbons or oils.     

Production of a Bioemulsifier with Potential Application in the Food Industry

Biosurfactants are of considerable interest due to their biodegradability, low degree of toxicity, and diverse applications. However, the high production costs involved in the acquisition of biosurfactants underscore the need for optimization of the production process to enable viable application on an industrial scale. The aims of the present study were to select a species of Candida that produces a biosurfactant with the greatest emulsifying potential and to investigate the influence of components of the production medium and cultivation conditions. Candida utilis achieved the lowest surface tension (35.53 mN/m), best emulsification index (73 %), and highest yield (12.52 g/l) in a medium containing waste canola frying oil as the carbon source and ammonium nitrate as the nitrogen source. The best combination of medium components and cultivation conditions was 6 % (w/v) glucose, 6 % (w/v) waste canola frying oil, 0.2 % (w/v) ammonium nitrate, 0.3 % (w/v) yeast extract, 150 rpm, 1 % inoculum (w/v), and 88 h of fermentation. The greatest biosurfactant production and the lowest surface tension were achieved in the first 24 h of production, and the maximum biomass production was recorded at 72 h. The biosurfactant produced from C. utilis under the conditions investigated in the present study has a potential to be a bioemulsifier for application in the food industry.     

Comparative study of biosurfactant produced by microorganisms isolated from formation water of petroleum reservoir

Biosurfactant produced by Pseudomonas aeruginosa, Bacillus subtilis and Rhodococcus erythropolis that isolated from the formation water of Chinese petroleum reservoir has been compared in surface abilities and oil recovery. Maximum biosurfactant production reached to about 2.66 g/l and the surface tension of liquid decreased from 71.2 to 22.56 mN/m using P. aeruginosa. Three strains exhibited a good ability to emulsify the crude oil, and biosurfactant of P. aeruginosa attained an emulsion index of 80% for crude oil which was greater than other strains. Stability studies were carried out under the extreme environmental conditions, such as high temperature, pH, salinity and metal ions. Results showed an excellent resistance of all biosurfactants to retain their surface-active properties at extreme conditions. It was found that the biosurfactants from three isolated bacteria showed a good stability above pH of 5, but at lower pH (from 1 to 5) they will harmfully be affected. They were able to support the condition up to 20 g/l salinity. P. aeruginosa biosurfactant was even stable at the higher salinity. Regarding temperature, all produced biosurfactants demonstrated a good stability in the temperature up to 120 °C. But stability of three biosurfactants was affected by monovalent and trivalent ions. Oil recovery experiments in physical simulation showed 7.2-14.3% recovery of residual oil after water flooding when the biosurfactant of three strains was added. These results suggest that biosurfactants of these indigenous isolated strains are appropriate candidates for enhanced oil recovery with a preference to biosurfactant of P. aeruginosa.     

Utilization of Banana Peel as a Novel Substrate for Biosurfactant Production by Halobacteriaceae archaeon AS65

In this study, biosurfactant-producing bacteria was evaluated for biosurfactant production by using banana peel as a sole carbon source. From the 71 strains screened, Halobacteriaceae archaeon AS65 produced the highest biosurfactant activity. The highest biosurfactant production (5.30 g/l) was obtained when the cells were grown on a minimal salt medium containing 35 % (w/v) banana peel and 1 g/l commercial monosodium glutamate at 30 °C and 200 rpm after 54 h of cultivation. The biosurfactant obtained by extraction with ethyl acetate showed high surface tension reduction (25.5 mN/m), a small critical micelle concentration value (10 mg/l), thermal and pH stability with respect to surface tension reduction and emulsification activity, and a high level of salt tolerance. The biosurfactant obtained was confirmed as a lipopeptide by using a biochemical test FT-IR, NMR, and mass spectrometry. The crude biosurfactant showed a broad spectrum of antimicrobial activity and had the ability to emulsify oil, enhance PAHs solubility, and oil bioremediation.     

Exopolysaccharides and Antimicrobial Biosurfactants Produced by Paenibacillus macerans TKU029

Paenibacillus macerans TKU029 can produce exopolysaccharides (EPSs; 3.46 g/L) and a biosurfactant (1.78 g/L) in a medium with 2 % (w/v) squid pen powder as the sole carbon/nitrogen source. The biosurfactant can reduce the surface tension of water from 72.30 to 35.34 mN/m at a concentration of 2.76 g/L and reach an emulsification index of 56 % after a 24-h reaction with machine oil. This biosurfactant is stable at 121 °C for 20 min, over a pH range from 3 to 11, and in <5 % salt solutions. It also shows significant antimicrobial activity, which remains active after treatment at 121 °C and at pH values from 4 to 10, against Escherichia coli BCRC13086, Staphylococcus aureus BCRC10780, Fusarium oxysporum BCRC32121 and Aspergillus fumigatus BCRC30099. Furthermore, human skin shows from 37.3 to 44.3 % hydration after being treated with TKU029 EPSs for 180 min. These results imply that EPSs and the biosurfactant from this strain have potential in cosmetics, for removal of oil contamination, and as antimicrobial agents.     

Antimicrobial and Antibiofilm Activity of Designed and Synthesized Antimicrobial Peptide, KABT-AMP

Lysine-rich peptide, designated as KABT-AMP, was designed and synthesized to supersede the irrational use of chemical antibiotics as standard therapy. KABT-AMP is a 22-amino acid helical cationic peptide (+10) and amphipathic in nature. The antimicrobial kinetics of the peptide was ascertained in the representative strains of gram-positive, gram-negative, and fungal strains, viz., Staphylococcus aureus MTCC 2940, Escherichia coli MTCC 2939, and Candida albicans MTCC 227, respectively. KABT-AMP was synthesized by solid-phase synthesis and purified using reverse-phase high-performance liquid chromatography which resulted in >95 % purity, and matrix-assisted laser desorption/ionization time of flight revealed the mass of the peptide to be 2.8 kDa. KABT-AMP showed significant broad-spectrum antimicrobial activity against the bacterial and fungal strains analyzed in the present study with survivability of 30.8, 30.6, and 31.7 % in E. coli, S. aureus, and C. albicans, respectively, at 6 h. KABT-AMP also demonstrated antibiofilm activity against the tested biofilm forming clinical isolate, Candida tropicalis. The putative membranolytic activity of the peptide was substantiated by electron microscopic analysis. Results reveal that KABT-AMP will exhibit noteworthy antimicrobial activity against multidrug-resistant bacteria and fungus at micromolar concentrations with minimal cytotoxicity and thus could be conceived for biomedical application.     

Microbial surfactant-enhanced mineral oil recovery under laboratory conditions

Microbial enhanced oil recovery (MEOR) is potentially useful to recover incremental oil from a reservoir being beyond primary and secondary recovery operations. Effort has been made to isolate and characterize natural biosurfactant produced by bacterial isolates collected from various oil fields of ONGC in Assam. Production of biosurfactant has been considered to be an effective major index for the purpose of enhanced oil recovery. On the basis of the index, four promising bacterial isolates: Pseudomonas aeruginosa (MTCC7815), P. aeruginosa (MTCC7814), P. aeruginosa (MTCC7812) and P. aeruginosa (MTCC8165) were selected for subsequent testing. Biosurfactant produced by the promising bacterial isolates have been found to be effective in the recovery of crude oil from saturated column under laboratory conditions. Two bacterial strains: P. aeruginosa (MTCC7815) and P. aeruginosa (MTCC7812) have been found to be the highest producer of biosurfactant. Tensiometer studies revealed that biosurfactants produced by these bacterial strains could reduce the surface tension (sigma) of the growth medium from 68 to 30 mN m(-1) after 96 h of growth. The bacterial biosurfactants were found to be functionally stable at varying pH (2.5-11) conditions and temperature of 100 degrees C. The treatment of biosurfactant containing, cell free culture broth in crude oil saturated sand pack column could release about 15% more crude oil at 90 degrees C than at room temperature and 10% more than at 70 degrees C under laboratory condition.     

Bacteriocin PJ4 Active Against Enteric Pathogen Produced by Lactobacillus helveticus PJ4 Isolated from Gut Microflora of Wistar Rat (Rattus norvegicus): Partial Purification and Characterization of Bacteriocin

The increase of multidrug-resistant pathogens and the restriction on the use antibiotics due to its side effects have drawn attention to the search for possible alternatives. Bacteriocins are small antimicrobial peptides produced by numerous bacteria. Much interest has been focused on bacteriocins because they exhibit inhibitory activity against pathogens. Lactic acid bacteria possess the ability to synthesize antimicrobial compounds (like bacteriocin) during their growth. In this study, an antibacterial substance (bacteriocin PJ4) produced by Lactobacillus helveticus PJ4, isolated from rat gut microflora, was identified as bacteriocin. It was effective against wide assay of both Gram-positive and Gram-negative bacteria involved in various diseases, including Escherichia coli, Bacillus subtilis, Pseudomonas aeruginosa, Enterococcus faecalis, and Staphylococcus aureus. The antimicrobial peptide was relatively heat-resistant and also active over a wide pH range of 2–10. It has been partially purified to homogeneity using ammonium sulfate precipitation and size exclusion chromatography and checked on reverse-phase high-performance liquid chromatography. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis of bacteriocin PJ4 purified through size exclusion chromatography resolved ~6.5 kDa protein with bacteriocin activity. The peptide is inactivated by proteolytic enzymes, trypsin, and lipase but not when treated with catalase, α-amylase, and pepsin. It showed a bactericidal mode of action against the indicator strains E. coli MTCC443, Lactobacillus casei MTCC1423, and E. faecalis DT48. Such characteristics indicate that this bacteriocin may be a potential candidate for alternative agents to control important pathogens.     

Evaluation of Cashew Apple Juice for Surfactin Production by Bacillus subtilis LAMI008

Bacillus subtilis LAMI008 strain isolated from the tank of Chlorination at the Wastewater Treatment Plant on Campus do Pici in Federal University of Ceará, Brazil has been screened for surfactin production in mineral medium containing clarified cashew apple juice (MM-CAJC). Results were compared with the ones obtained using mineral medium with glucose PA as carbon source. The influence on growth and surfactin production of culture medium supplementation with yeast extract was also studied. The substrate concentration analysis indicated that B. subtilis LAMI008 was able to degrade all carbon sources studied and produce biosurfactant. The highest reduction in surface tension was achieved with the fermentation of MM-CAJC, supplemented with yeast extract, which decreased from 58.95?±?0.10 to 38.10?±?0.81 dyn cm^?1. The biosurfactant produced was capable of emulsifying kerosene, achieving an emulsification index of 65%. Surfactin concentration of 3.5 mg L^?1 was obtained when MM-CAJC, supplemented with yeast extract, was used, thus indicating that it is feasible to produce surfactin from clarified cashew apple juice, a renewable and low-cost carbon source.     

Biotechnological Production of Phenyllactic Acid and Biosurfactants from Trimming Vine Shoot Hydrolyzates by Microbial Coculture Fermentation

Coculture fermentations show advantages for producing food additives from agroindustrial wastes, considering that different specified microbial strains are combined to improve the consumption of mixed sugars obtained by hydrolysis. This technology dovetails with both the growing interest of consumers towards the use of natural food additives and with stricter legislations and concern in developed countries towards the management of wastes. The use of this technology allows valorization of both cellulosic and hemicellulosic fractions of trimming vine shoots for the production of lactic acid (LA), phenyllactic acid (PLA), and biosurfactants (BS). This work compares the study of the potential of hemicellulosic and cellulosic fractions of trimming vine shoots as cheaper and renewable carbon sources for PLA and BS production by independent or coculture fermentations. The highest LA and PLA concentrations, 43.0 g/L and 1.58 mM, respectively, were obtained after 144 h during the fermentation of hemicellulosic sugars and simultaneous saccharification and fermentation (SSF) carried out by cocultures of Lactobacillus plantarum and Lactobacillus pentosus. Additionally, cell-bond BS decreased the surface tension (ST) in 17.2 U; meanwhile, cell-free supernatants (CFS) showed antimicrobial activity against Salmonella enterica and Listeria monocytogenes with inhibition halos of 12.1?±?0.6 mm and 11.5?±?0.9 mm, respectively.     

Evaluation of Anthocyanin Content,Antioxidant Potential and Antimicrobial Activity of Black,Purple and Blue Colored Wheat Flour and Wheat-Grass Juice against Common Human Pathogens

The present study aimed to analyze the antioxidant and antimicrobial activity of anthocyanins extracted from colored wheat flour and wheat-grass juice against human pathogens. The total anthocyanin content and antioxidant potential in colored wheat flour and wheat-grass juice extracts were significantly higher than white flour and wheat-grass juice extracts. Ultra-performance liquid chromatography showed the maximum number of anthocyanin peaks in black wheat, with delphinidin-3-o-galactoside chloride, delphinidin-3-o-glucoside chloride, and cyanindin-3-o-glucoside chloride as the major contributors. Among flour extracts, maximum zones of inhibition against Staphylococcus aureus (MTCC 1934), Pseudomonas aeruginosa (MTCC 1434), Escherichia coli, and Candida albicans (MTCC 227) were produced by black flour extract, having the highest anthocyanin content. It exhibited a minimum microbicidal concentration (MMC) of 200 mg/mL against E. coli and C. albicans; and 100 and 150 mg/mL against S. aureus and P. aeruginosa, respectively. Black and purple flour extracts exhibited a minimum inhibitory concentration (MIC) of 50 mg/mL against S. aureus and P. aeruginosa. White flour extracts did not show MMC against E. coli and C. albicans. Among wheat-grass juice extracts, black wheat-grass was most effective and showed an MIC of 100–150 mg/mL against all pathogens. It exhibited an MMC of 200 mg/mL against S. aureus and P. aeruginosa. Hence, anthocyanin-rich colored wheat could be of nutraceutical importance.     

Green synthesis of novel quinoline based imidazole derivatives and evaluation of their antimicrobial activity

We have described the conventional and microwave method for the synthesis of N-(4-((2-chloroquinolin-3-yl)methylene)-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)(aryl)amides 3a–l. It is observed that the solvent-free microwave thermolysis is a convenient, rapid, high-yielding, and environmental friendly protocol for the synthesis of quinoline based imidazole derivatives when compared with conventional reaction in a solution phase. Antimicrobial activity of the newly synthesized compounds is screened in vitro on the following microbial cultures: Escherichia coli (MTCC 443), Pseudomonas aeruginosa (MTCC 1688), Staphylococcus aureus (MTCC 96), Streptococcus pyogenes (MTCC 442), Candida albicans (MTCC 227), Aspergillus niger (MTCC 282), Aspergillus clavatus (MTCC 1323). All the synthesized bio-active molecules are tested for their in vitro antimicrobial activity by bioassay namely serial broth dilution. Among these compounds 3c, 3d, 3f, 3h and 3j show significant potency against different microbial strains. All the compounds have been characterized by IR, ¹H NMR, ¹³C NMR and mass spectral data. On the basis of statistical analysis, it is observed that these compounds give significant co-relation.     

Cytotoxic Effect of Microbial Biosurfactants Against Human Embryonic Kidney Cancerous Cell: HEK-293 and Their Possible Role in Apoptosis

Two different microbial biosurfactants S9BS and CHBS were isolated from Lysinibacillus fusiformis S9 and Bacillus tequilensis CH. Cytotoxicity effect of these biosurfactants on human embryonic kidney cancerous cell (HEK-293) were studied with the help of 3-(4,5-dimethylthiazol-2yl-)-2, 5-diphenyl tetrazolium bromide (MTT) assay and morphological changes were observed under inverted microscope. The biosurfactants exhibited positive cytotoxic effect on HEK-293 cell line. It was found that LC₅₀ of S9BS and CHBS were 75 and 100 μg ml^?1, respectively. Further cell cycle and apoptosis analysis of biosurfactant-treated HEK-293 cell line were done by FACS. In this study, cytotoxic effect of glycolipid biosurfactant against HEK-293 cell lines is reported for the first time. Mechanism towards increased membrane permeability of biosurfactant-treated cancer cell may be the incorporation of its lipid moiety into the plasma membrane leading to formation of pores and membrane disruption. Hence, these microbial biosurfactants can prove to be significant biomolecule for cancer treatment.     

Evaluation of waste products in the synthesis of surfactants by yeasts

The highest yields of biosurfactants were obtained by: (i) Pseudozyma antarctica (107.2 g L^?1) cultivated in a medium containing post-refining waste; (ii) Pseudozyma aphidis (77.7 g L^?1); and (iii) Starmerella bombicola (93.8 g L^?1) both cultivated in a medium with soapstock; (iv)Pichia jadinii (67.3 g L^?1) cultivated in a medium supplemented with waste frying oil. It was found that the biosurfactant synthesis yield increased in all strains when the cell surface hydrophobicity reached 70–80 %, enabling the microbial cells to make good contact with hydrophobic substrates. The lowest surface tension of the post-cultivation medium was from 32.0 mN m^?1 to 37.8 mN m^?1. However, this parameter (which was also determined by a drop collapse assay) was of limited use in monitoring biosurfactant synthesis in this study. The crude glycerol was not a good substrate for biosurfactant synthesis although, in the case of P. aphidis, 67.4 g L^?1 of biosurfactants were obtained after cultivation in the medium supplemented with glycerol fraction (GF2). In a low-cost medium containing soapstock and whey permeate or molasses, about 90 g L^?1 of mannosylerythritol lipids were synthesised by P. aphidis and approximately 40 g L^?1 by P. antarctica.     

Application of Lipopeptide Biosurfactant Isolated from a Halophile: Bacillus tequilensis CH for Inhibition of Biofilm

Biosurfactants are amphiphilic molecules having hydrophobic and hydrophilic moieties produced by various microorganisms. These molecules trigger the reduction of surface tension or interfacial tension in liquids. A biosurfactant-producing halophile was isolated from Lake Chilika, a brackish water lake of Odisha, India (19°41′39″N, 85°18′24″E). The halophile was identified as Bacillus tequilensis CH by biochemical tests and 16S rRNA gene sequencing and assigned accession no. KC851857 by GenBank. The biosurfactant produced by B. tequilensis CH was partially characterized as a lipopeptide using thin-layer chromatography, Fourier transform infrared spectroscopy, and nuclear magnetic resonance techniques. The minimum effective concentration of a biosurfactant for inhibition of pathogenic biofilm (Escherichia coli and Streptococcus mutans) on hydrophilic and hydrophobic surfaces was found to be 50 μg ml^?1. This finding has potential for a variety of applications.     

Synthesis,characterization, antimicrobial evaluation and QSAR studies of organotin(IV) complexes of Schiff base ligands of 2-amino-6-substituted benzothiazole derivatives

A series of organotin(IV) complexes of type R₂SnLCl [R = Ph, Bu, Et, Me] were prepared by reaction of diorganotindichloride(IV) with Schiff base ligands, L₁ = (1-[(6-ethoxy-benzothiazol-2-ylimino)-methyl]-naphthalen-2-ol), L₂ = (1-[(6-nitro-benzothiazol-2-ylimino)-methyl]-naphthalen-2-ol), L₃ = (1-[(6-methoxy-benzothiazol-2-ylimino)-methyl]-naphthalen-2-ol) and L₄ = (1-[(6-methyl-benzothiazol-2-ylimino)-methyl]-naphthalen-2-ol) obtained from 2-amino-6-substituted benzothiazole derivatives with 2-hydroxy-1-naphthaldehyde in 1:1 molar ratio. These organotin(IV) complexes were characterized by various spectroscopic techniques (¹H, ¹³C and ¹¹⁹Sn NMR, FT-IR), and physical techniques (X-ray powder diffraction analysis and elemental analysis). The coordination of the prepared complexes has been planned as pentacoordinated around the central tin atom during which ligands coordinated to tin atom in bidentate manner acted as N, O donor system. The ligands and their complexes were screened for antibacterial and antifungal activities against Gram-positive bacteria Bacillus cereus (MTCC 10072), Staphylococcus aureus (NCIM 2901), Gram-negative bacteria Escherichia coli (MTCC 732), Pseudomonas aeruginosa (MTCC 424) and fungi Aspergillus niger (MTCC 9933) and Aspergillus flavus (ATCC 76801). The output of QSAR analysis indicated that topological parameters (molecular connectivity indices) were responsible for controlling the antimicrobial activity of the synthesized compounds.     

Lipopeptide surfactant production by Bacillus subtilis grown on low-cost raw materials

The production of biosurfactant by Bacillus subtilis ATCC 6633 was investigated using commercial sugar, sugarcane juice and cane molasses, sugarcane juice alcohol stillage, glycerol, mannitol, and soybean oil. Commercial sugar generated the minimum values of surface tension, with the best results (28.7 mN/m, (relative critical micelle concentration [CMC⁻¹] of 78.6) being achieved with 10 g of substrate/L in 48 h. At a pH between 7.0 and 8.0, a higher production of surface-active compounds and a greater emulsifier activity was also observed. Enrichment of the culture medium with trace minerals and EDTA showed maximum yields, whereas supplementation with yeast extract stimulated only cell growth. The kinetic studies revealed that biosurfactant production is a cell growth-associated process; surface tension, CMC, and emulsification index values of 29.6 dyn/cm, 82.3, and 57%, respectively, were achieved, thus indicating that it is feasible to produce biosurfactants from a renewable and low-cost carbon source.     

Production of Microbial Rhamnolipid by Pseudomonas Aeruginosa MM1011 for Ex Situ Enhanced Oil Recovery

Recently, several investigations have been carried out on the in situ bacteria flooding, but the ex situ biosurfactant production and addition to the sand pack as agents for microbial enhanced oil recovery (MEOR) has little been studied. In order to develop suitable technology for ex situ MEOR processes, it is essential to carry out tests about it. Therefore, this work tries to fill the gap. The intention of this study was to investigate whether the rhamnolipid mix could be produced in high enough quantities for enhanced oil recovery in the laboratory scale and prove its potential use as an effective material for field application. In this work, the ability of Pseudomonas aeruginosa MM1011 to grow and produce rhamnolipid on sunflower as sole carbon source under nitrogen limitation was shown. The production of Rha-C₁₀-C₁₀ and Rha₂-C₁₀-C₁₀ was confirmed by thin-layer chromatography and high-performance liquid chromatography analysis. The rhamnolipid mixture obtained was able to reduce the surface and interfacial tension of water to 26 and 2 mN/m, respectively. The critical micelle concentration was 120 mg/L. Maximum rhamnolipid production reached to about 0.7 g/L in a shake flask. The yield of rhamnolipid per biomass (Y _RL/x ), rhamnolipid per sunflower oil (Y _RL/s ), and the biomass per sunflower oil (Y _x/s ) for shake flask were obtained about 0.01, 0.0035, and 0.035 g g^?1, respectively. The stability of the rhamnolipid at different salinities, pH and temperature, and also, its emulsifying activity has been investigated. It is an effective surfactant at very low concentrations over a wide range of temperatures, pHs, and salt concentrations, and it also has the ability to emulsify oil, which is essential for enhanced oil recovery. With 120 mg/L rhamnolipid, 27 % of original oil in place was recovered after water flooding from a sand pack. This result not only suggests rhamnolipids as appropriate model biosurfactants for MEOR, but it even shows the potential as a biosurfactant of choice for actual MEOR applications.     

First Report of a Lipopeptide Biosurfactant from Thermophilic Bacterium Aneurinibacillus thermoaerophilus MK01 Newly Isolated from Municipal Landfill Site

A biosurfactant-producing thermophile was isolated from the Kahrizak landfill of Tehran and identified as a bacterium belonging to the genus Aneurinibacillus. A thermostable lipopeptide-type biosurfactant was purified from the culture medium of this bacterium and showed stability in the temperature range of 20–90 °C and pH range of 5–10. The produced biosurfactant could reduce the surface tension of water from 72 to 43 mN/m with a CMC of 1.21 mg/mL. The strain growing at a temperature of 45 °C produces a substantial amount of 5 g/L of biosurfactant in the medium supplemented with sunflower oil as the sole carbon source. Response surface methodology was employed to optimize the biosurfactant production using sunflower oil, sodium nitrate, and yeast extract as variables. The optimization resulted in 6.75 g/L biosurfactant production, i.e., 35 % improved as compared to the unoptimized condition. Thin-layer chromatography, FTIR spectroscopy, ¹H-NMR spectroscopy, and biochemical composition analysis confirmed the lipopeptide structure of the biosurfactant.