Evaluation of Robustness of the Biosurfactant Derived from Balanites aegyptiaca (L.) Del.

Balanites aegyptiaca (L.) Del. is a widely distributed xerophytic multipurpose tree. The mesocarp of the fruit of B. aegyptiaca has detergent properties due to the presence of saponins. The stability potential of this biosurfactant at varying pH, temperature, and salinity has not been explored so far. In the present study, the relative surface tensions of five different concentrations of the biosurfactant were studied at different temperatures, salinity, and under pH conditions. This study reveals that this biosurfactant retains its activity over a wide range of pH (3–11) and at high salinity (7% NaCl). It is a thermostable cationic surfactant; surfactant activity was recorded even at 100 °C with the lowest relative surface tension of 0.47. High oil displacement (18.00 mm) was observed when studied with petrol. This biosurfactant was found to have a high emulsification index (E₂₄) of 70% with mustard oil. These results indicate that biosurfactant derived from B. aegyptiaca may find use in a wide range of sectors such as textile, food, cosmetics, oil recovery, and healthcare under a wide range of physical and chemical conditions. It offers an efficient, economically viable, and plant-derived alternative to synthetic detergents and adds a way to maintain a sustainable environment.     

Enrichment and Separation of Sinomenine and Acutumine from Sinomenium acutum by pH-Zone-Refining Counter-Current Chromatography

Enrichment and separation of alkaloids from a chloroform extract of Sinomenium acutum has been successfully performed for the first time using pH-zone-refining counter-current chromatography. The two-phase solvent system used for enrichment was composed of Methyl tert-butyl ether (MtBE)–acetonitrile (CH₃CN)–water (4:1:5, v/v), where 10 mM triethylamine (TEA) was added to the upper organic stationary phase as a retainer and 10 mM hydrochloric acid (HCl) to the aqueous mobile phase as an eluter, which could enrich the alkaloids from the crude extract well. For the preparative separation, the solvent system consisted of MtBE–CH₃CN–water (4:0.5:5, v/v) with 10 mM TEA in organic stationary phase and 5 mM HCl in the aqueous mobile phase, which could separate and purify the enriched crude alkaloids successfully. 0.82 g of crude alkaloids was enriched from 1.60 g of chloroform extract in the first step separation. From the enriched crude alkaloids, 376 mg of sinomenine and 85 mg of acutumine were obtained in the second step separation with the purity of 98.1% and 98.7%, respectively. The chemical structures of the isolated compounds were identified by UV, ESI-MS and ¹H NMR.     

Achieving the Best Yield in Glycolipid Biosurfactant Preparation by Selecting the Proper Carbon/Nitrogen Ratio

Pseudomonas aeruginosa RS29, the native biosurfactant-producing strain isolated from the oil fields of Assam, India was used to investigate the influence of the carbon nitrogen ratio on production of the biosurfactant. The biosurfactant producing ability of the strain was measured based on surface tension (ST) reduction of the culture medium and the emulsification (E24) index. Production was greatly influenced by the sources of nitrogen and carbon as well as the carbon to nitrogen (C/N) ratio. Sodium nitrate was the best nitrogen source and the water miscible carbon source, glycerol was observed as the best carbon source for maximum biosurfactant production. The C/N ratio 12.5 allowed the maximum production of biosurfactant by the RS29 strain. At this C/N ratio, 55 % ST of the culture medium was reduced by the produced biosurfactant. Concentrations of crude and rhamnolipid biosurfactant obtained at this particular C/N ratio were 5.6 and 0.8 g/l respectively. The RS29 strain was novel as it was able to produce a sufficient amount of biosurfactant utilizing a much lower amount of the water miscible carbon source, glycerol. Extraction of the biosurfactant by a chloroform–methanol (2:1) mixture was the best method to obtain the highest biosurfactant from the culture medium of the strain. The biosurfactant was confirmed as a mixture of mono and di-rhamnolipid congeners, Rha–C₁₀–C₁₀–CH₃ being the most abundant one. The biosurfactant was a good foaming and emulsifying agent.     

Production of surface-active sophorolipid biosurfactant and crude oil degradability by novel Rhodotorula mucilaginosa strain SKF2

Biosurfactants are produced by important types of microorganisms such as bacteria, yeast, and filamentous fungi and have been used in a variety of industries. Among the 15 crude oil-degrading fungi, the two molds and one yeast were identified by 18S rDNA sequences as Mucor circinelloides strain SKMC, Fusarium fujikuroi strain DB2, and Rhodotorula mucilaginosa strain SKF2. These strains were isolated from crude oil–contaminated soil, diesel oil–contaminated soil, and activated sludge in the Oil Refinery Plant in Isfahan, Iran, respectively. The yeast strain was identified as a novel crude oil–degrading and biosurfactant-producing fungi in the presence of (1% v/v) Iranian light crude oil in the minimal salt medium (MSM). The highest amount of the dry weight of produced biosurfactant was measured at 6.2 g L⁻¹. Chemical nature of produced biosurfactant was determined as a surface-active sophorolipid biosurfactant compound by thin-layer chromatography, Fourier transform infra-red spectroscopy, and gas chromatography–mass spectrometry (GC–MS) analysis. The residual hydrocarbons in the MSM were analyzed by GC–MS, and it was shown that octadecane and docosane were eliminated by this novel strain completely.     

Separation and Purification of Three High-Purity Isoflavonoids from Belamcanda chinensis (L.) DC. by Supercritical Fluid Extraction and High-Speed Counter-Current Chromatography

Supercritical fluid extraction (SFE) was used to extract three isoflavonoids including irigenin, irisfloretin and dichtomitin from Belamcanda chinensis (L.) DC. The parameters including pressure, temperature, sample particle size, and flow rate of CO₂ were optimized with an orthogonal test. Under the optimized conditions of 15 MPa, 55°C, a sample particle size of 20–40 mesh and CO₂ flow rate of 40 L h^?1. The process was then scaled up by 10 times using a preparative SFE system. The yield of the crude extract from SFE was 4.1%, which contained irigenin, irisfloretin, and dichtomitin 0.71%, 0.49%, and 0.05%, respectively. To compare the extraction methods, Soxhlet Extraction (SE) was performed. The results indicated that SFE was better than SE. Irigenin, irisfloretin, and dichtomitin in the SFE extract were then separated and purified by high-speed counter-current chromatography (HSCCC) with a two-phase solvent system composed of petroleum ether–ethyl acetate–methanol–water (2:4:3:3, v/v). From 5.0 g of dry crude extract, 27.8 mg irigenin, 16.4 mg irisfloretin, and 2.1 mg dichtomitin were obtained at purities of 97.1%, 96.4%, and 98.0%, respectively, as determined by HPLC-PDA. These results well indicate that SFE and HSCCC are very powerful techniques for the extraction and purification of irigenin, irisfloretin, and dichtomitin from B. chinensis.     

Biosurfactant Production by Bacillus tequilensis ZSB10: Structural Characterization,Physicochemical, and Antifungal Properties

A new biosurfactant was obtained from a moderately halophilic bacterium identified as Bacillus tequilensis ZSB10 that was isolated from a saline water pond located in Tehuacan-Cuicatlan valley, Mexico. A kinetic analysis of the bacterial growth of the ZSB10 strain showed a maximum growth at 24 h regardless of the initial pH (5, 7.4, and 9). The best results were found at pH = 7.4 in terms of bacterial growth, besides which the produced biosurfactant showed emulsifying and surfactant properties with an emulsification index (E₂₄) and surface tension change (ΔST) of 54 ± 0% and 26 mN m⁻¹, respectively. Extracted ZSB10 crude biosurfactant had a yield of 106 ± 6 mg L⁻¹, an E₂₄ = 58.4 ± 0.2%, and a ΔST = 26 mN m⁻¹ with a critical micelle concentration (CMC) of 44.82 mg L⁻¹. Also, its structure was characterized by MALDI-TOF mass spectrometry as a surfactin, iturin A, and fengycin mixture whose main isoform was leu/ile-7 C₁₅ surfactin [M + Na]⁺. Finally, the ZSB10 crude biosurfactant showed antifungal activity against Helminthosporium sp., with a 79.3% growth inhibition and an IC₅₀ of 1.37 mg per disc. Therefore, this biosurfactant could be used as biopesticide.     

Chemical Analysis of Polyphenols with Antioxidant Capacity from Pinus durangensis Bark

The most active phenolics in Pinus durangensis residual bark were identified and evaluated following a chromatographic fractionation. Bark powder was defatted with hexane, and a crude extract (CE) was obtained by extraction with aqueous acetone (67%). A liquid partition with ethyl acetate was performed to produce an organic extract (OE), which was subsequently purified by column chromatography (Toyopearl HW-40F, methanol), resulting in ten fractions (MF1 to MF10) and an oligomeric fraction eluted with acetone 67% (OLF). Subfraction MF6-1 was obtained by a second chromatographic purification of MF6. Extraction yields, total phenolics, flavonoids, and flavanols contents were determined in CE and OE. The antioxidant activity of bark extracts was measured by DPPH and ABTS assays at 100 µg/mL, expressed in percentage, median effective concentration (IC₅₀), and TEAC (mM). Also the low density lipoprotein inhibition was evaluated. Identification of major phenolics was carried out by HPLCESI–MS and HPLC–DAD instruments. Bioactive taxifolin (dihydroquercetin), dihydromyricetin, myricetin, quercetin, pinomyricetin (myricetin-methoxy), pinoquercetin (quercetin-methoxy), trimeric, and tetrameric procyanidins were detected and identified in P. durangensis bark extracts. Polyphenols found are similar to those contained in Pycnogenol and other Pinus species.     

Studies on Biosurfactants Produced using Bacillus cereus Isolated from Seawater with Biotechnological Potential for Marine Oil-Spill Bioremediation

With the aim of producing a biotensioactive material for use in the remediation of marine environments, screening for biosurfactant-producing bacteria was conducted with strains isolated from seawater contaminated with petroleum derivatives. Gene sequencing revealed that all four promising biosurfactant-producing isolates belonged to the same genus and species, namely Bacillus cereus. The biosurfactant-producing bacteria were cultivated with different carbon (glucose, soybean oil, and waste frying soybean oil) and nitrogen (ammonium chloride, sodium nitrate, urea, and peptone) sources. B. cereus strain BCS0 was chosen as the best biosurfactant producer in a mineral medium with 2% frying oil and 0.12% peptone. Following the optimization of agitation and cultivation time, an agitation rate of 250 rpm and 48 h of cultivation were selected. Under these conditions, the surface tension was reduced to 27 mN m⁻¹ and the biosurfactant concentration was 3.5 g L⁻¹. The critical micelle concentration (CMC) of the biosurfactant was defined as 500 mg L⁻¹. The biosurfactant remained stable within large ranges of pH (2–10), salinity (2–10%), and temperature (5–120 °C). Under these conditions, motor oil emulsification rates were greater than 90%. Moreover, the biosurfactant properties remained unaltered after heating at 90 °C for 120 min. The biosurfactant enhanced the degradation of motor oil up to 96% in 27 days and exhibited considerable motor oil displacement capacity. Thus, the biosurfactant has potential in the application of remediation processes in marine environments.     

Multifactorial Approach to Biosurfactant Production by Adaptive Strain Candida tropicalis MTCC 230 in the Presence of Hydrocarbons

In this study, Candida tropicalis MTCC 230 was used to adapted in hydrocarbon along with glucose for biosurfactant production, showing diauxic growth during the production. Biosurfactant was characterized through TLC and FTIR analysis as surfactin, a lipopeptide. Process parameters were optimized one factor at a time, showing the highest emulsification index (%E₂₄) at 54 %. The production of biosurfactant was enhanced by using biostatistically based experimental design with the interactive effect of different parameters. On the basis of Placket–Burman design, four factors, hydrocarbon, ammonium chloride, microelements and temperature are found to be significant (P < 0.05) for the production of biosurfactant. A second order polynomial regression model in central composite design estimated the maximum biosurfactant production in terms of the emulsification index (%E₂₄). The optimum combination of different parameters for the biosurfactant production, obtained for hydrocarbon, ammonium chloride, microelements and temperature are 81.41 %, 1.63 (g/l), 1.69 (g/l) and 35.25 °C, respectively. The biosurfactant production was increased twofold after optimization and selection of interactive parameters by response surface methodology.     

Simultaneous recovery of glycyrrhizic acid and liquiritin from Chinese licorice root (Glycyrrhiza uralensis Fisch) by aqueous two-phase system and evaluation biological activities of extracts

Aqueous two-phase system (ATPS) was used for simultaneous purification of glycyrrhizic acid (GA) and liquiritin (LQ) from crude extract of Chinese licorice root. It was revealed that 87% GA and 94% LQ were retrieved in the ATPS top phase, under the optimum conditions of 25% (w/w) ethanol, 30% (w/w) K₂HPO₄ and 4% (w/w) loading sample at 10–40°C. Compared with crude extract, the ATPS top-phase extract exhibited the highest antioxidative activity, but no tyrosinase inhibitory effect. Whereas, the ATPS bottom-phase extract was proved to be effective ABTS radical scavenger and tyrosinase inhibitor, suggesting the potency of the alcohol-salt ATPS purification for the different medicinal purposes.     

Biosurfactant-Assisted Bioremediation of Polycyclic Aromatic Hydrocarbons (PAHs) in Liquid Culture System and Substrate Interactions

A lipopeptide biosurfactant was produced by the bacterium Pseudomonas aeruginosa strain LBP9 isolated from petroleum-contaminated soil. Phenanthrene, fluoranthene, and pyrene were used as model polycyclic aromatic hydrocarbons (PAHs) to study the effect of the biosurfactant on the biodegradation of mixed and sole substrate PAHs, and examine substrate interactivity effects on their biodegradation in liquid culture. At 400 mg/L amendment of lipopeptide, the solubility of phenanthrene, fluoranthene, and pyrene were increased to 19, 33, and 45 times their aqueous solubility, respectively, and the extent of substrate utilization rate (q_max?) of PAHs was enhanced up to three-fold in the sole substrate studies in comparison to the unamended controls. In the ternary PAH mixture at total concentration of 300 mg/L, with equal parts of each PAH, 77%, 57%, and 33% degradation of phenanthrene, fluoranthene, and pyrene were observed, respectively, at 400 mg/L lipopeptide amendment on day 30 of incubation. Whereas in the sole substrate experiments at 300 mg/L concentration of each PAH and the same level of lipopeptide amendment more than 98% fluoranthene and 76% pyrene were degraded and phenanthrene removal was so rapid that at day 4 of incubation more than 80% was degraded. Biosurfactants at optimum amounts enhanced biodegradation of PAHs. Lipopeptide amendments of 200 mg/L and 400 mg/L were found out to be optimum amounts for statistically significant (p < 0.05) biodegradation of the PAHs in the experiments. However, despite biosurfactant-enhanced bioavailability of the PAHs, biodegradation rate was competitively inhibited in the multisubstrate microcosms.     

Effect of Trichilia americana Extract on Feeding Behavior of Asian Armyworm, Spodoptera litura

We investigated the desensitization of Asian armyworms, Spodoptera litura, to a crude methanolic extract of wood Trichilia americana. Following repeated exposures to the extract over four days, desensitization was observed under no-choice conditions at low extract concentration (0.5 g/cm²). No desensitization was observed under choice conditions or at a higher concentration (5.0 g/cm²). Repeated exposures of larvae to the extract over 7.5 hr showed that larvae increasingly tolerated the extract due to hunger, rather than showing true desensitization. When larvae were exposed to the extract in test arenas of different sizes, larvae showed reduced feeding and desensitization as arena size increased. Finally, treatment of whole cabbage plants with extract resulted in larvae abandoning the plant. The crude extract was effective at protecting the plants, although desensitization did not occur, because the larvae moved away from the plant.     

The Role of Pyoluteorin from Pseudomonas protegens Pf-5 in Suppressing the Growth and Pathogenicity of Pantoea ananatis on Maize

The rhizospheric bacterium Pseudomonas protegens Pf-5 can colonize the seed and root surfaces of plants, and can protect them from pathogen infection. Secondary metabolites, including lipopeptides and polyketides produced by Pf-5, are involved in its biocontrol activity. We isolated a crude extract from Pf-5. It exhibited significant surface activity and strong antibacterial activity against Pantoea ananatis DZ-12, which causes maize brown rot on leaves. HPLC analysis combined with activity tests showed that the polyketide pyoluteorin in the crude extract participated in the suppression of DZ-12 growth, and that the lipopeptide orfamide A was the major biosurfactant in the crude extract. Further studies indicated that the pyoluteorin in the crude extract significantly suppressed the biofilm formation of DZ-12, and it induced the accumulation of reactive oxygen species in DZ-12 cells. Scanning electron microscopy and transmission electron microscopy observation revealed that the crude extract severely damaged the pathogen cells and caused cytoplasmic extravasations and hollowing of the cells. The pathogenicity of DZ-12 on maize leaves was significantly reduced by the crude extract from Pf-5 in a dose-dependent manner. The polyketide pyoluteorin had strong antibacterial activity against DZ-12, and it has the potential for development as an antimicrobial agent.     

Merits and limitations of TiO<Subscript>2</Subscript>-based photocatalytic pretreatment of soils impacted by crude oil for expediting bioremediation

Heavy hydrocarbons (HHCs) in soils impacted by crude oil spills are generally recalcitrant to biodegradation due to their low bioavailability and complex chemical structure. In this study, soils were pretreated with varying concentrations of ultraviolet radiation A (UVA) or ultraviolet radiation C (UVC) activated titanium dioxide (TiO₂) (1%–5%) under varying moisture conditions (0%–300% water holding capacity (WHC)) to enhance biodegradation of HCCs and shorten remediation timeframes. We demonstrate that pretreatment of impacted soils with UVC-activated TiO₂ in soil slurries could enhance bioremediation of HHCs. Total petroleum hydrocarbon (TPH) removal after 24 h exposure to UVC (254 nm and 4.8 mW/cm²) was (19.1 ± 1.6)% in slurries with 300% WHC and 5 wt-% TiO₂. TPH removal was non-selective in the C15-C36 range and increased with moisture content and TiO₂ concentration. In a 10-d bioremediation test, TPH removal in treated soil increased to (26.0 ± 0.9)%, compared to (15.4 ± 0.8)% for controls without photocatalytic pre-treatment. Enhanced biodegradation was also confirmed by respirometry. This suggests that addition of UVC-activated TiO₂ to soil slurries can transform recalcitrant hydrocarbons into more bioavailable and biodegradable byproducts and increase the rate of subsequent biodegradation. However, similar results were not observed for soils pretreated with UVA activated TiO₂. This suggests that activation of TiO₂ by sunlight and direct addition of TiO₂ to unsaturated soils within landfarming setting may not be a feasible approach. Nevertheless, less than 1% of UVA (7.5 mW/cm²) or UVC (1.4 mW/cm²) penetrated beyond 0.3 cm soil depth, indicating that limited light penetration through soil would hinder the ability of TiO₂ to enhance soil bioremediation under land farming conditions.

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Analysis of the Activated Sludge Process in an MBR under Starvation Conditions

An aerobic membrane bioreactor (MBR) at complete biomass retention was studied over a period of time under starvation conditions. Kinetic parameters were determined in a no‐feed batch test. The decay rate of activated sludge, k_d = 0.05 d^–1, was determined by tracking the decrease of MLSS. The ratio of MLVSS/MLSS was in the range 0.76–0.85. The pH values were between 7.02 and 8.23. As a function of different initial concentrations of MLSS, specific nitrification rates q_N, decreased from 4.23 to 0.02 mg‐N/(g MLVSS d) and specific biodegradation rates q_b increased from 0.23 to 1.90 mg‐COD/(g MLVSS d). From experimental data the kinetic constants for respiration, which followed Monod kinetics, were determined as q_O2max = 9.8 mg‐O₂/(g MLVSS h), K_x = 2.9 g/dm³. Additionally, a linear correlation between MLSS and mean floc size was found to exist during the biodegradation process.     

Screening and identification of Pseudomonas aeruginosa AB4 for improved production,characterization and application of a glycolipid biosurfactant using low‐cost agro‐based raw materials

BACKGROUND: The study is focused on (i) screening and taxonomic identity of a bacterial strain for biosurfactant production, and (ii) evaluation of its potential for production of a biosurfactant using agro‐based feedstock(s) and characterization of it for application in the removal of heavy metals. RESULTS: The production of biosurfactant by an isolate Pseudomonas aeruginosa AB4 (identified on the basis of 16S rRNA analysis) using various cost‐effective substrates were examined at conditions 40 °C, 120 rpm for 7 days. It revealed maximum (40 gL^?1) rhamnolipids production and 46% reduction of initial surface tension. Its optimum production was achieved at (i) C:N ratio 10:0.6, (ii) pH 8.5 and (iii) 40 °C. The cell–free supernatant examined for biosurfactant activity by (i) haemolytic assay, (ii) CTAB‐ methylene blue assay, (iii) drop collapse test, (iv) oil spreading technique and (v) EI 24 assay showed its glycolipid nature and stable emulsification. Analysis of partially purified rhamnolipids by (i) thin layer chromatography (TLC), (ii) high performance thin layer chromatography (HPTLC), (iii) high performance liquid chromatography (HPLC), (iv) Fourier transform infrared (FT‐IR) and (v) gas chromatography–mass spectrometry (GC‐MS) confirmed its structure as methyl ester of 3‐hydroxy decanoic acid (a glycolipid) with two major structural congeners (Rha‐C₁₀‐C₁₀ and Rha‐C₁₀‐C₈) of mono‐rhamnolipids. Finally, it showed sequestration of Cd and Pb, suggesting its application in biosurfactant‐assisted heavy metal bioremediation. CONCLUSION: This work has screened and identified a bacterium with superior biosurfactant production capabilities, characterized the glycolipidic biosurfactants as rhamnolipid and indicated the feasibility of biosurfactant production using novel renewable, relatively inexpensive and easily available resources such as non‐edible vegetable de‐oiled seed cakes and showed its utility in remediation of heavy metals. Copyright © 2010 Society of Chemical Industry     

Characterization of Biosurfactant Produced by a Novel Strain of Pseudomonas aeruginosa,Isolate ADMT1

Several biosurfactant‐producing bacterial strains were isolated from petroleum‐contaminated soil. The isolate ADMT1, identified as a new strain of Pseudomonas aeruginosa, was selected for further studies on the basis of oil displacement test and emulsification index (E24). The optimal parameters for production, determined by employing Box–Behnken design, were temperature 36.5 °C and pH 7. The environmental isolate ADMT1 produced significant amount of biosurfactant (1.7 g L^?1 in 72 h) in minimal salt medium (MSM) using dextrose as the sole carbon source. The E24 value and critical micelle concentration (CMC) of the biosurfactant was 100% and 150 mg L^?1, respectively. At CMC, the surface tension of water was reduced to 28.4 mN m^?1. The biosurfactant exhibited hemolytic activity and antibacterial activity against 8 reference strains of pathogenic bacteria, including 2 methicillin‐resistant Staphylococcus aureus strains (MRSA ATCC 562 and MRSA ATCC 43300), with minimum inhibitory concentration (MIC) of 0.4 and 0.2 mg mL^?1, respectively. The structure of biosurfactant was characterized by FTIR, ¹H, and ¹³C NMR spectroscopy. 7 di‐rhamnolipid (RL) congeners were identified in the biosurfactant by ultraperformance liquid chromatography–mass spectrometry analysis. The major congeners, which constituted 67% of the RL mixture, included Rha‐Rha‐C₁₀‐C₁₀, Rha‐Rha‐C₁₂‐C₁₀, and Rha‐Rha‐C_12:1‐C₁₀. The minor congeners were Rha‐Rha‐C₁₀‐C₈, Rha‐Rha‐C_10:1‐C₁₀, Rha‐Rha‐C₁₀‐C_14:1, and Rha‐Rha‐C₁₀‐C₁₄. The congener Rha‐Rha‐C₁₀‐C₁₄ is being reported for the first time from any species of Pseudomonas. The high surface activity and E24 value make the ADMT1‐RL a potential candidate for its use in detergents, environmental bioremediation, and as an emulsifier in the food industry.     

APPLICATION OF POWER LAW LOGISTIC MODEL TO GROWTH KINETICS OF BACILLUS LICHENIFORMIS MS3 ON A WATER-INSOLUBLE SUBSTRATE

The power law logistic model was utilized to investigate the growth of a hydrocarbon assimilating bacterium on a water-insoluble substrate. To achieve this end, population dynamics of Bacillus licheniformis MS3 in a medium containing n-decane as the sole carbon source was monitored for 30 h. Different initial biosurfactant concentrations and shaking rates were employed to examine the role of mass transfer in the cell growth and the consequent hydrocarbon biodegradation. The amount of n-decane degraded in the system was detected by gas chromatography at the end of the incubation period. The results revealed that when mass transfer limitations were lessened through addition of an initial biosurfactant concentration and agitation, the bacterial growth increased more than three times and the n-decane biodegradation was enhanced from 6.7 to 15.1 mg/100 mL. Finally, the power law logistic model proved to be highly capable in simulating both the experimental results and various systems with water-insoluble carbon sources.     

Biomonitoring of biosurfactant production by green fluorescent protein‐marked Bacillus subtilis W1012

BACKGROUND: Biosurfactant production was investigated using two strains of Bacillus subtilis, one being a reference strain (B. subtilis 1012) and the other a recombinant of this (B. subtilis W1012) made able to produce the green fluorescent protein (GFP). RESULTS: Batch cultivations carried out at different initial levels of glucose (G₀) in the presence of 10 g L^?1 casein demonstrated that the reference strain was able to release higher levels of biosurfactants in the medium at 5.0≤G₀≤10 g L^?1 (B_max = 104–110 mg L^?1). The recombinant strain exhibited slightly lower levels of biosurfactants (B_max = 90–104 mg L^?1) but only at higher glucose concentrations (G₀ ≥ 20 g L^?1). Under these nutritional conditions, the fluorescence intensity linked to the production of GFP was shown to be associated with the cell concentration even after achievement of the stationary phase. CONCLUSION: The ability of the genetically‐modified strain to simultaneously overproduce biosurfactant and GFP even at low biomass concentration makes it an interesting candidate for use as a biological indicator to monitor indirectly the biosurfactant production in bioremediation treatments. Copyright © 2008 Society of Chemical Industry     

Structural characterization of a biosurfactant produced by Bacillus subtilis at 45°C

Structural and biochemical characterization of a biosurfactant produced by Bacillus subtilis under thermophilic conditions was performed. Preliminary structural determination of CHCl₃/CH₃OH (65∶15) extracts by thin-layer chromatographic reagents showed it to be identical to surfactin. Also, the infrared, ¹H nuclear magnetic resonance, and mass spectroscopy analysis confirmed it to be identical to surfactin. Biochemically, the surfactant was a lipopeptide-containing lipid (17.05%) and protein (13.2%). The surfactant yielded a minimal aqueous surface-tension value of 28 dyne/cm and an interfacial tension value at 0.1% concentration of 0.2 dyne/cm against diesel oil. The critical micelle concentration of the surfactant was 35 mg/L. The biosurfactant exhibited an emulsification value (E ₂₄) of 90 against diesel oil and a sand-pack oil recovery of 62%. It has potential application in microbial-enhanced oil recovery in thermophilic, alkaline, acidic, and halophilic environments.