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.     

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.     

Low-Toxic and Nonirritant Biosurfactant Surfactin and its Performances in Detergent Formulations

Developing eco-friendly, nonirritant, low-toxic, and high-efficient surface active ingredients for detergents is an ongoing challenge in the detergent field. Surfactin is one of the representative lipopeptides produced by microorganisms. In this article, we report the surfactin isolated from cell-free broth of Bacillus subtilis HSO121 and purified by reversed-phase high-performance liquid chromatography for detergent formulations. The biodegradability, acute dermal irritation, acute oral toxicity (LD₅₀ and LC₅₀), surface activity, washing efficiency, and compatibility with hard water of the purified biosurfactant surfactin have been studied to explore the feasibility for applications of the surfactin in detergents. Acute oral toxicity tests (LD₅₀ > 5000 mg kg⁻¹, LC₅₀ > 1000 mg kg⁻¹) and skin irritation tests (PII = 0) indicate that the surfactin is a low-toxic and nonirritant ingredient for detergent formulation. Moreover, the surfactin shows excellent surface and interfacial properties of emulsification and wettability, high compatibility, and stability in a wide range of temperatures, pH, and hard water and acceptable properties in biodegradability and foaming ability, which suggests that the biosurfactant surfactin is a promising ingredient for detergent formations in our daily life and for industrial applications.     

Batch production of biosurfactant with foam fractionation

Methods of producing the biosurfactant surfactin from cultures of Bacillus subtilis (BBK006) have been investigated. A reactor with integrated foam fractionation was designed and used in batch mode, and the performance compared with that of the same culture in shaken flasks. In the batch reactor, significant foaming occurred between 12.5 h and 14.5 h of culture time. During this period, the foam was routed through the foam fractionation column to a mechanical foam breaker, and a biosurfactant‐enriched foamate was collected. Concentration of surfactin in the foamate product was around 50 times greater than that in the culture medium. Using the integrated reactor, 136 mg L^?1 of surfactin was produced, significantly more than was achieved in shaken flasks (92 mg L^?1). The foam fractionation method allowed a real‐time measurement of the rate of surfactin production during growth. This showed that the maximum rate of production occurred at the interphase between log and stationary modes of growth, in contrast to previous work showing that surfactin is exclusively a secondary metabolite. The high value of surfactin yield in relation to biomass (Y_P/x = 0.262) indicated that surfactin was produced very efficiently by Bacillus subtilis (BBK006) in this integrated bioreactor. Copyright © 2006 Society of Chemical Industry     

Surfactin生物表面活性剂及其驱油研究进展

表面活性素（surfactin）是一类由革兰氏阳性的枯草芽孢杆菌产生的脂肽（lipopeptide）型生物表面活性剂,因其具有优于化学合成表面活性剂的若干优点,如低毒性、高生物降解性、更好的环境相容性,且在极端环境下稳定性好,在提高石油采收率方面有较好的应用潜力,但是目前只有少数的生物表面活性剂可以大规模生产实现工业化应用。本文介绍了surfactin生物表面活性剂的化学结构和生物合成机制,并对其发酵生产过程的影响因素进行分析,为提高其生产经济性探索不同的策略,例如使用更便宜的原材料、优化培养基组分、优化反应器等,系统论述了surfactin生物表面活性剂的驱油机理和其与化学合成表面活性剂的复配研究,同时针对其应用时的不足之处提出研究新思路。     

Culture Medium Evaluation Using Low-Cost Substrate for Biosurfactants Lipopeptides Production by Bacillus amyloliquefaciens in Pilot Bioreactor

Total biosurfactant production by Bacillus amyloliquefaciens IT45 was evaluated with different substrates concentrations in a culture medium. A central composite design (CCD) was developed to evaluate the influence of variables, including glucose syrup, yeast extract, and calcium chloride, on surface tension (ST), total biosurfactant production, and residual sugar (RS). As a result, the best observed results for ST, RS, and total biosurfactant production were 30 mN m⁻¹, 31%, and 5.5 g L⁻¹, respectively, after 48 h of fermentations carried out in batch operation process. Characterization of the biosurfactant identified the presence of surfactin. To validate the CCD experiments, fermentations were conducted in a 40 L pilot bioreactor. For this fermentation, the cellular growth was 3.0 × 10⁹ CFU mL⁻¹, surfactin production was 0.55 g L⁻¹, and RS was 28%. The results demonstrate that B. amyloliquefaciens IT45 has the potential to produce biosurfactants and does not require high concentrations of carbon and nitrogen sources for its development.     

Continuous production of biosurfactant with foam fractionation

A glucose‐limited chemostat was used to determine the growth parameters of BBK006 for continuous production of the biosurfactant surfactin. The continuous cultivation exhibited low maintenance metabolism (m = 0.39 mmol_glucose g_bacteria^?1 h^?1) and high molar growth yield ( g_bacteria mol_glucose^?1). It was found that the surfactin production rate in continuous culture was not only a function of dilution rate but also varied with the initial concentration of glucose in the feed. A high steady state concentration of surfactin (18 mg L^?1) was maintained in the culture at a dilution rate of 0.2 h^?1 when glucose concentration in the feed was 0.25 g L^?1. This is the first demonstration of continuous surfactin production and recovery using glucose as a carbon source. The production of surfactin is known to be related to the age of the microorganisms and a simple mathematical model has been constructed to show how the age‐related production can be quantified. Copyright © 2006 Society of Chemical Industry     

Sustainable Production of Biosurfactant from Agro-Industrial Oil Wastes by Bacillus subtilis and Its Potential Application as Antioxidant and ACE Inhibitor

The microbial conversion of agro-industrial oil wastes into biosurfactants shows promise as a biomass refinery approach. In this study, Bacillus subtilis #309 was applied to produce surfactin using rapeseed and sunflower cakes, the most common oil processing side products in Europe. Studies of the chemical composition of the substrates were performed, to determine the feasibility of oil cakes for surfactin production. Initially, screening of proteolytic and lipolytic activity was performed to establish the capability of B. subtilis #309 for substrate utilization and hence effective surfactin production. B. subtilis #309 showed both proteolytic and lipolytic activity. The process of surfactin production was carefully analyzed by measurement of the surfactin concentration, pH, surface tension (ST) and emulsification index (E₂₄). The maximal surfactin concentration in the sunflower and rapeseed cake medium reached 1.19 ± 0.03 and 1.45 ± 0.09 g/L, respectively. At the same time, a progressive decrease in the surface tension and increase in emulsification activity were observed. The results confirmed the occurrence of various surfactin homologues, while the surfactin C₁₅ was the dominant one. Finally, the analysis of surfactin biological function exhibited antioxidant activity and significant angiotensin-converting enzyme (ACE)-inhibitory activity. The half-maximal inhibitory concentration (IC₅₀) value for ACE inhibition was found to be 0.62 mg/mL for surfactin. Molecular docking of the surfactin molecule to the ACE domains confirmed its inhibitory activity against ACE. Several interactions, such as hydrophobic terms, hydrogen bonds and van der Waals interactions, were involved in the complex stabilization. To the best of our knowledge, this is the first report describing the effect of a lipopeptide biosurfactant, surfactin, produced by B. subtilis for multifunctional properties in vitro, namely the ACE-inhibitory activity and the antioxidant properties, using different assays, such as 2,2-azinobis (3-ethyl-benzothiazoline-6-sulfonic acid (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP). Thus, the ACE-inhibitory lipopeptide biosurfactant shows promise to be used as a natural antihypertensive agent.     

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.     

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.     

Biosurfactants Production Using Permeate from Whey Ultrafiltration and Bioproduct Recovery by Membrane Separation Process

The management of whey is a challenge for dairy products where the volume produced is remarkable. This problem is minimized through membrane separation processes (MSP) to obtain whey protein concentrate, which has high added value. However, a permeate effluent stream is still generated that is composed of lactose, vitamins, and minerals, which can serve as raw material for the production of biotechnological compounds. Thus, this study aimed to produce biosurfactants using the permeate from whey ultrafiltration as part of the culture media of the bioprocess, to recover the biosurfactant produced using MSP, and to identify the biocompound. The production was carried out using Bacillus methylotrophicus and Bacillus pumilus. The variables nitrogen source (urea or ammonium sulfate), nitrogen source concentration (0.5% or 1.0%), inducer (soybean oil or biodiesel), inducer concentration (1% or 2%), and the addition of micronutrients (with our without) were studied using a fractional factorial experimental design 2^5-1_IV. In the fermentation processes, it was possible to verify the biosurfactant production through the reduction of surface tension, obtaining a minimum value of 35.07 mN/m for B. methylotrophicus and 26.02 mN/m for B. pumilus. Recovery via MSP was an efficient strategy for biosurfactant purification, which was concentrated in the fraction of the retentate. We produced a high-value-added biocompound identified as surfactin, valuing the permeate residue from whey ultrafiltration.     

Effects of various nutritional supplements on biosurfactant production by a strain of <Emphasis Type="Italic">Bacillus subtilis</Emphasis> at 45°C

Effects of various factors on growth and biosurfactant production by Bacillus subtilis MTCC 2423 were studied. Sucrose (2%) and potassium nitrate (0.3%) were the best carbon and nitrogen sources. The addition of various metal supplements (magnesium, calcium, iron, and trace elements) greatly affected growth and biosurfactant production. The effect of the metal cations, used together, is greater than when they are used individually. The biosurfactant production increased considerably (almost double) by addition of metal supplements. Very high concentrations of metal supplements, however, inhibited biosurfactant production. Amino acids such as aspartic acid, asparagine, glutamic acid, valine, and lysine increased the final yield of biosurfactant by about 60%. The organism could produce biosurfactant at 45°C and within the pH range of 4.5–10.5. The biosurfactant was thermostable and pH stable (from 4.0 to 12.0). The capability of the organism to produce biosurfactant under thermophilic, alkaliphilic, and halophilic conditions makes it a suitable candidate for field applications. Infrared, nuclear magnetic resonance, and mass spectroscopy studies showed the surfactant to be identical to surfactin.     

Volumetric Oxygen Mass Transfer Coefficient and Surface Tension in Simulated Salt Bioremediation Media

Toluene can be removed from contaminated sites via bioremediation through the addition of biosurfactant compounds, which reduce the surface tension. However, aeration and mixing must be optimized to ensure an effective volumetric oxygen mass transfer coefficient (k_La). Experiments were perfomed with different salt containing solutions, which were tested either as such, or with different supplements. k_La values obtained at different agitation intensities and aeration rates were compared with those in water, and correlated with power number and superficial gas velocity. Surface tension decreased when surfactin was added to toluene‐containing media. The seawater‐simulating medium exhibited the highest surface tension reduction.     

Benchmarking the Self‐Assembly of Surfactin Biosurfactant at the Liquid–Air Interface to those of Synthetic Surfactants

The adsorption of surfactin, a lipopeptide biosurfactant, at the liquid–air interface has been investigated in this work. The maximum adsorption density and the nature and the extent of lateral interaction between the adsorbed surfactin molecules at the interface were estimated from surface tension data using the Frumkin model. The quantitative information obtained using the Frumkin model was also compared to those obtained using the Gibbs equation and the Langmuir–Szyszkowski model. Error analysis showed a better agreement between the experimental and the calculated values using the Frumkin model relative to the other two models. The adsorption of surfactin at the liquid–air interface was also compared to those of synthetic anionic, sodium dodecylbenzenesulphonate (SDBS), and nonionic, octaethylene glycol monotetradecyl ether (C₁₄E₈), surfactants. It has been estimated that the area occupied by a surfactin molecule at the interface is about 3‐ and 2.5‐fold higher than those occupied by SDBS and C₁₄E₈ molecules, respectively. The interaction between the adsorbed molecules of the anionic biosurfactant (surfactin) was estimated to be attractive, unlike the mild repulsive interaction between the adsorbed SDBS molecules.     

Surfactant Effects on Aeration Performance of Stirred Tank Reactors

The effect of surfactants on aeration performance in stirred tank reactors (STR) at high rates of foaming is studied. The volumetric oxygen transfer coefficient (k_La) and foaming activity estimated as foaming height (H_f) were determined. Biotechnology of lipopeptide biosurfactants from aerobic organisms, e.g., Bacillus subtilis were addressed. Using model solutions of known foam‐generating properties, high‐molecular weight surfactin and low‐molecular weight sodium dodecyl sulphate (SDS), as well as impellers of different types, with flat and fluid‐foil blades, clues on the concentration dependence of STR oxygen transfer and foaming as well as options for foam reduction in the presence of biosurfactant were sought. In response to a two‐fold decrease of surface tension by surfactin, k_La values decreased up to 30 % but remained within the range expected for the mixing system in water; the experiments with SDS showing stronger dependence on surfactant concentration and surface tension. Mixing of surfactant media by a standard six‐blade disc turbine (RT) imposed rate limitations on gassing. A low‐shear impeller Narcissus (NS) could be used to avoid bulk foam outflow, while preserving k_La values that remained unchanged. The ‘power per unit volume' correlation of k_La in stirred tanks is tested in the presence of surfactin.     

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     

Response Surface Optimization of the Critical Media Components for the Production of Surfactin

Optimization of the fermentation media for maximization of surfactin production was carried out. The carbon source (glucose), the nitrogen source (ammonium nitrate) and the mineral salts ferrous and manganous sulphates were the critical components of the medium optimized. A 2⁴ full factorial central composite experimental design followed by multi-stage Monte-Carlo optimization was used in the design of experiments and in the analysis of results. This procedure limited the number of actual experiments performed while allowing for possible interactions between the four components. The optimum values for the tested variables for the maximal production of surfactin were (in g dm⁻³): glucose = 36·5; NH₄NO₃ = 4·5; FeSO₄ = 4×10⁻³ and MnSO₄ = 27·5 ×10⁻². Relative surfactant concentrations were expressed as the reciprocal of the critical micelle concentration (CMC⁻¹) and the maximum predicted yield of surfactin in terms of CMC⁻¹ was 45·5. © 1997 SCI.     

FTIR Characterization of Polymorphic Transformation of Ammonium Nitrate

Solid ammonium nitrate (NH ₄ NO ₃ ) exists in five stable polymorphic forms (designated as phases I, II, III, IV, and V) below its melting point at around 170°C. Phase IV is stable in a temperature range of ?17°C ～ 32°C and is the only phase that has been considered by the atmospheric research community until recently. In this study, we examine the IV ? III phase transition of NH ₄ NO ₃ and how relative humidity (RH) affects the transition path and the transition temperatures using in-situ microscopic Fourier Transform InfraRed spectroscopy. Two kinds of NH ₄ NO ₃ samples, powder produced from grinding commercially produced chemicals and single particles obtained by efflorescence of droplets on PTFE filters, were studied. The powder samples exhibit the IV?III phase transition and the transition temperature depends on the RH while the single particle samples exhibit only the IV?II transition at about 52°C (forward) and 48°C (reverse), bypassing phase III, with transition temperatures independent of the RH. However, grinding of the particles produced through efflorescence results in the IV?III transitions. Differences in crystal structure and moisture content may explain the distinct phase transition behaviors of the two types of samples. These results suggest that solid and pure NH ₄ NO ₃ aerosol particles are stable in phase IV under ambient conditions.     

Effect of early hydration temperature on hydration product and strength development of magnesium phosphate cement (MPC)

This paper investigates the effect of early hydration temperatures on hydration products and strength development of magnesium phosphate cement (MPC). MPC paste specimens with borate contents of 3%, 6%, 9% and 12% are prepared and cured in different air temperatures and in the adiabatic condition. The internal hydration temperatures are measured by pre-embedded temperature probes. MPC samples with different hydration temperatures are also obtained by using thin slice samples. The hydration products in MPC samples with different hydration temperatures are analyzed by X-ray diffractometer (XRD) and scanning electron microscope (SEM) and the strength development is also measured. The results show that NH₄MgPO₄·6H₂O is the major hydration product and beneficial to strength development of MPC at hydration temperature below 70 °C. NH₄MgPO₄·H₂O is another major product, which significantly decreases the strength, when the temperature is higher than a critical temperature between 70 °C and 75 °C. NH₄MgPO₄·H₂O can directly form in the MPC paste, and comes from the decomposition of NH₄MgPO₄·6H₂O when the temperature is above 75 °C. With temperature elevation and duration extension, NH₄MgPO₄·6H₂O decomposes rapidly, and even entirely when the temperature is above 100 °C. The borate content has no effect on the types of major hydration products and the critical temperature.     

An Investigation of Catalysts for the On Board Synthesis of NH3. A Possible Route to Low Temperature NOx Reduction for Lean-Burn Engines

Platinum group metal catalysts have been investigated for the formation of NH₃ from NO + H₂ at low temperatures in the absence and presence of CO. Although CO inhibits the formation of NH₃, substantial amounts are still observed with a Pt catalyst. By combining Pt with a support (ceria–zirconia) that has low temperature NO_x storage characteristics it has been shown in transient experiments that NH₃ can be formed and stored in situ under rich conditions, and may then be used to reduce NO_x under lean burn conditions.