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.     

Synthesis,Characterization, and Oil Recovery Application of Biosurfactant Produced by Indigenous <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> WJ-1 Using Waste Vegetable Oils

A bacterial strain was isolated and cultured from the oil excavation areas in tropical zone in northern China. The biochemical characteristics and partial sequenced 16S rRNA gene of isolate, WJ-1, was identical to those of cultured representatives of the species Pseudomonas aeruginosa. This bacterium was able to produce a type of biosurfactant. Compositional analysis revealed that the extracted biosurfactant was composed of high percentage lipid (∼74%, w/w) and carbohydrate (∼20%, w/w) in addition to a minor fraction of protein (∼6%, w/w). The best production of 50.2 g/l was obtained when the cells were grown on minimal salt medium containing 6.0% (w/v) glucose and 0.75% (w/v) sodium nitrate supplemented with 0.1% (v/v) element solution at 37 °C and 180 rpm after 96 h. The optimum biosurfactant production pH value was found to be 6.0–8.0. The biosurfactant of WJ-1, with the critical micelle concentration of 0.014 g/L, could reduce surface tension to 24.5 mN/m and emulsified kerosene up to EI₂₄ ≈95. The results obtained from time course study indicated that the surface tension reduction and emulsification potential was increased in the same way to cell growth. However, maximum biosurfactant production occurred and established in the stationary growth phase (after 90 h). Thin layer chromatography, Fourier transform infrared spectrum, and mass spectrum analysis indicate the extracted biosurfactant was affiliated with rhamnolipid. The core holder flooding experiments demonstrated that the oil recovery efficiency of strain and its biosurfactant was 23.02% residual oil.     

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.     

Enhanced Production of Bacterial Cellulose by Using Gluconacetobacter hansenii NCIM 2529 Strain Under Shaking Conditions

Bacterial cellulose (BC), a biopolymer, due to its unique properties is valuable for production of vital products in food, textile, medicine, and agriculture. In the present study, the optimal fermentation conditions for enhanced BC production by Gluconacetobacter hansenii NCIM 2529 were investigated under shaking conditions. The investigation on media components and culture parameters revealed that 2 % (w/v) sucrose as carbon source, 0.5 % (w/v) potassium nitrate as nitrogen source, 0.4 % (w/v) disodium phosphate as phosphate source, 0.04 % (w/v) magnesium sulfate, and 0.8 % (w/v) calcium chloride as trace elements, pH?5.0, temperature 25 °C, and agitation speed 170 rpm with 6 days of fermentation period are optimal for maximum BC production. Production of BC using optimized media components and culture parameters was 1.66 times higher (5.0 g/l) than initial non optimized media (3.0 g/l). Fourier transform infrared spectroscopy spectrum and comparison with the available literature suggests that the produced component by G. hansenii in the present study is pure bacterial cellulose. The specific action of cellulase out of the investigated hydrolytic enzymes (cellulase, amylase, and protease) further confirmed purity of the produced BC. These findings give insight into conditions necessary for enhanced production of bacterial cellulose, which can be used for a variety of 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.     

Three-Phase Partitioning as a Rapid and Easy Method for the Purification and Recovery of Catalase from Sweet Potato Tubers (Solanum tuberosum)

Three-phase partitioning (TPP) was used to purify and recover catalase from potato crude extract. The method consists of ammonium sulfate saturation, t-butanol addition, and adjustment of pH, respectively. The best catalase recovery (262 %) and 14.1-fold purification were seen in the interfacial phase in the presence of 40 % (w/v) ammonium sulfate saturation with 1.0:1.0 crude extract/t-butanol ratio (v/v) at pH 7 in a single step. The sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the enzyme showed comparatively purification and protein molecular weight was nearly found to be 56 kDa. This study shows that TPP is a simple, economical, and quick method for the recovering of catalase and can be used for the purification process.     

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 Microbial Cellulose by a Bacterium Isolated from Fruit

This study presents the production of bacterial cellulose (BC) by a bacterium isolated from a rotten fruit and its process optimization. Here, isolation and screening of potent cellulose producers were carried out from different natural sources, viz., soil, rotten fruits, and vegetables and vinegar. A total of 200 bacterial isolates were obtained, which were screened for cellulose production using Hestrin?CSchramm medium. A novel and potent cellulose-producing bacterium was newly isolated from a rotten fruit and identified as Gluconacetobacter sp. F6 through 16S ribosomal DNA sequencing and morphological, cultural, and biochemical characteristics. After optimization of culture conditions, including pH, temperature, agitation, carbon/nitrogen sources, and inducers, the BC production was greatly increased from 0.52 to 4.5?g/l (8.65-fold increase). The optimal culture medium contained 1% (w/v) glucose, 1.5% (w/v) yeast extract, 0.5% (w/v) peptone, 0.27% (w/v) disodium hydrogen phosphate, 0.115% (w/v) citric acid, and 0.4% (w/v) ethanol. BC produced was analyzed for the presence of cellulose fibrils by epiflourescent microscopy using Calcofluor white stain and scanning electron microscopy and confirmed by NMR. There are very scanty reports about the optimization of BC production by bacteria isolated from rotten fruits.     

In Situ Biphasic Extractive Fermentation for Hexanoic Acid Production from Sucrose by Megasphaera elsdenii NCIMB 702410

Hexanoic acid production by a bacterium using sucrose as an economic carbon source was studied under conditions in which hexanoic acid was continuously extracted by liquid–liquid extraction. Megasphaera elsdenii NCIMB 702410, selected from five M. elsdenii strains, produced 4.69 g l^?1 hexanoic acid in a basal medium containing sucrose. Production increased to 8.19 g l^?1 when the medium was supplemented by 5 g l^?1 sodium butyrate. A biphasic liquid–liquid extraction system with 10 % (v/v) alamine 336 in oleyl alcohol as a solvent was evaluated in a continuous stirred-tank reactor held at pH 6. Over 90 % (w/w) of the hexanoic acid in a 0.5 M aqueous solution was transferred to the extraction solvent within 10 h. Cell growth was not significantly inhibited by direct contact of the fermentation broth with the extraction solvent. The system produced 28.42 g l^?1 of hexanoic acid from 54.85 g l^?1 of sucrose during 144 h of culture, and 26.52 and 1.90 g l^?1 of hexanoic acid was accumulated in the extraction solvent and the aqueous fermentation broth, respectively. The productivity and yield of hexanoic acid were 0.20 g l^?1 h^?1 and 0.50 g g^?1 sucrose, respectively.     

Strategies for Enhancing the Production of Penicillin G Acylase from Bacillus badius: Influence of Phenyl Acetic Acid Dosage

Bacillus badius isolated from soil has been identified as potential producer of penicillin G acylase (PGA). In the present study, batch experiments performed at optimized inoculum size, temperature, pH, and agitation yielded a maximum PGA of 9.5 U/ml in shake flask. The experiments conducted in bioreactor with different oxygen flow rates revealed that 0.66 vvm oxygen flow rate could be sufficient for the maximum PGA activity of 12.7 U/ml. From a detailed investigation on the strategies of the addition of phenyl acetic acid (PAA) for increasing the production of PGA, it was found that the controlled addition of 10 ml of 0.1 % (w/v) PAA once in every 2 h from 6th hour of growth showed the maximum PGA activity of 32 U/ml. Thus, our studies for the first time showed that at concentration above 0.1 % (w/v) PAA, the PGA production decreased. This selective condition paves the way for less costly bioprocess for the production of PGA.     

Nonaqueous CE for Rapid and Sensitive Determination of Matrine and Oxymatrine in <Emphasis Type="Italic">Sophora flavescens</Emphasis> and Its Medicinal Preparations

A simple, rapid, and sensitive non-aqueous capillary electrophoresis procedure for the quantitative determination of matrine and oxymatrine is established. Optimum separation conditions were obtained when the sample was injected under pressure for 3 s at 50 mbar and separated with the buffer containing 70 mM ammonium acetate, 7.0% (v/v) acetic acid, and 10% (v/v) acetonitrile in methanol medium at 25 kV applied voltage. The analytes were detected at 205 nm. The two alkaloids can be separated within 12 min and quantified with high sensitivity. The method was validated in terms of reproducibility, linearity, and accuracy when applied to the analysis of matrine and oxymatrine in Sophora flavescens and its medicinal preparations.     

Optimization of Levan Production by Cold-Active <Emphasis Type="Italic">Bacillus licheniformis</Emphasis> ANT 179 and Fructooligosaccharide Synthesis by Its Levansucrase

Fructooligosaccharides (FOS) and levan attract much attention due to a wide range of applications in food technology and pharmaceutical and cosmetic industry. Bacillus licheniformis ANT 179, isolated from Antarctica soil, produced levansucrase and levan in a medium containing sucrose as carbon substrate. In this study, characterization of levansucrase and production of short-chain FOS and levan were investigated. Temperature and pH optimum of the enzyme were found to be 60 °C and pH 6.0, respectively. The optimization of fermentation conditions for levan production using sugarcane juice by response surface methodology (RSM) was carried out. Central composite rotatable design was used to study the main and the interactive effects of medium components: sugarcane juice and casein peptone concentration on levan production by the bacterium. The optimized medium with sugarcane juice at 20 % (v/v) and casein peptone at 2 % (w/v) was found to be optimal at an initial pH of 7.0 and incubation temperature of 35 °C for 48 h. Under these conditions, the maximum levan concentration was 50.25 g/L on wet weight basis and 16.35 g/L on dry weight basis. The produced inulin type FOS (kestose and neokestose) and levan were characterized by Fourier transform infrared spectroscopy (FT-IR) and nuclear magnetic resonance (NMR) analysis. The study revealed that the levansucrase could form FOS from sucrose. The locally available low-cost substrate such as sugarcane juice in the form of a renewable substrate is proposed to be suitable even for scale-up production of enzyme and FOS for industrial applications. The levan and FOS synthesized by the bacterium are suitable for food applications and biomedical uses as the bacterium has GRAS status and devoid of endotoxin as compared to other Gram-negative bacteria.     

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.     

Compositional Changes in Sugarcane Bagasse on Low Temperature, Long-term Diluted Ammonia Treatment

Sugarcane bagasse is the major by-product of the sugar industry. It has a great potential for the production of biofuels and chemicals due to its considerable amount of cellulose and hemicellulose. In this study, we investigated a simple and economic pretreatment process using dilute ammonia for the storage of sugarcane bagasse. Sugarcane bagasse was stored in 0, 0.03, and 0.3% (w/w) ammonium hydroxide in a closed bottle for 40 days at 30 °C under atmospheric pressure without any agitation or circulation. Samples were taken every 10 days and analyzed for changes on lignin, cellulose, hemicellulose composition, ammonia concentration, and microbial counts. Biomass storage for 40 days at 0.3% ammonium hydroxide removed 46% of lignin and retained 100% cellulose and 73% hemicellulose.     

Slow-Release Nutrient Capsules for Microorganism Stimulation in Oil Remediation

As the concern towards environmental deterioration grows worldwide, new technological achievements become essential for all countries. Among the technologies with great potential of bioremediation is microencapsulation of active material. Several studies have investigated the use of controlled release of active materials as a way of biostimulation and supplying the nutrients necessary for the bioremediation process. In fact, as the use of microorganisms has a great potential in degrading crude oils, this work aims to use that technology and to associate it to produce controlled-release capsules of nitrogen, phosphorus, and potassium (N, P, and K) for bioremediation purposes. For the capsule formulation, polymers of sodium alginate, Capsul®, and the commercial fertilizer NPK from Sempre Verde Inc. were used. Crude oil was the only carbon source and mineral medium for microorganism growth. Controlled-release nutrient capsules, with 4 mm in diameter, made of 3.0 % alginate (w/v) and 4.0 % Capsul® (w/v) were produced. Those capsules were used in association with a microbial consortium, in a liquid phase bioremediation process, having degraded 43.6 % of the total hydrocarbon within 240 h, evidencing thus as a promising tool for hydrocarbon bioremediation.     

Candida Biofilm Disrupting Ability of Di-rhamnolipid (RL-2) Produced from Pseudomonas aeruginosa DSVP20

Biosurfactant produced from Pseudomonas aeruginosa DSVP20 was evaluated for its potential to disrupt Candida albicans biofilm formed on polystyrene (PS) surfaces in this investigation. P. aeruginosa DSVP20 exhibited optimum production of biosurfactant (5.8 g?L^?1) after 96 h of growth with an ability to reduce surface tension of the aqueous solution from 72 to 28 mN?m^?1. Analysis of purified biosurfactant with FT-IR, ¹H and ¹³C NMR and MALDI-TOF MS revealed it to be di-rhamnolipid (RL-2) in nature. Biofilm disrupting ability of RL-2 (0.16 mg?mL^?1) on Candida cells when checked using XTT reduction assay revealed that about 50 % of the cells remain adhered to 96-well plate after 2 h of treatment, while up to 90 % reduction in pre-formed C. albicans biofilm on PS surface was observed with RL-2 (5.0 mg?mL^?1) in a dose-dependent manner. Microscopic analyses (SEM and CLSM) further confirm the influence of RL-2 on disruption of Candida biofilm extracellular matrix on PS surface which can be exploited as a potential alternative to the available conventional therapies.     

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.     

Ethanol Production from Starch Hydrolyzates using <Emphasis Type="Italic">Zymomonas mobilis</Emphasis> and Glucoamylase Entrapped in Polyvinylalcohol Hydrogel

The glucoamylase from Aspergillus niger, immobilized into poly(vinylalcohol) hydrogel lens-shaped capsules LentiKats®, was used for simultaneous saccharification and fermentation (SSF) with Zymomonas mobilis in free form. This system was stable in both the repeated batch and continuous mode of SSF. The microorganism was found to adsorb on the capsules with immobilized enzyme. This increased the ethanol productivity of the repeated batch system with 5% w/v of immobilized glucoamylase almost 2.1 times (7.2 g l^?1 h^?1) compared to free enzyme–free microorganism system (3.5 g l^?1 h^?1). The continuous SSF with the immobilized glucoamylase (11.5% w/v) tested for 15 days had productivity 10 g l^?1 h^?1, which is comparable to continuous experiments on semi-defined glucose medium (10 g l^?1 h^?1). These two systems were stable in both glucoamylase activity and microorganism productivity.     

Effectiveness of Sal Deoiled Seed Cake as an Inducer for Protease Production from Aeromonas sp. S1 for its Application in Kitchen Wastewater Treatment

The present study is an attempt to demonstrate the feasibility of sal (Shorea robusta) deoiled cake—a forest-based industrial by-product—as a cheaper media supplement for augmented protease production from Aeromonas sp. S1 and application of protease in the treatment of kitchen wastewater. Under optimized conditions, protease production could successfully be enhanced to 5.13-fold (527.5 U mL^?1) on using sal deoiled seed cake extract (SDOCE), as medium additive, compared to an initial production of 102.7 U mL^?1 in its absence. The culture parameters for optimum production of protease were determined to be incubation time (48 h), pH (7.0), SDOCE concentration (3 % (v/v)), inoculum size (0.3–0.6 % (v/v)), and agitation rate (100 rpm). The enzyme was found to have an optimum pH and temperature of 8.0 and 60 °C, respectively. The protease preparation was tested for treatment of organic-laden kitchen wastewater. After 96 h of wastewater treatment under static condition, enzyme preparation was able to reduce 74 % biological oxygen demand, 37 % total suspended solids, and 41 % oil and grease. The higher and improved level of protease obtained using sal deoiled seed cake-based media hence offers a new approach for value addition to this underutilized biomass through industrial enzyme production. The protease produced using this biomass could also be used as pretreatment tool for remediation of organic-rich food wastewater.     

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.