Optimization Of Biosurfactant Production Research Articles

Exploring the potential of novel Bacillus sp. G6: Isolation, characterization, and optimization of biosurfactant production from oil-contaminated soil

Biosurfactants (BSs) are highly biodegradable, environmentally benign, and multifunctional substances having diverse industrial uses, including environment bioremediation. Because of its remarkable emulsifying properties, as well as its use in both the environment and medicine, surfactin, is considered one of the most potent BSs and belongs to the lipopeptides family. The present study was performed to isolate the potential bacteria from oil-contaminated soils. Selected bacteria were checked for biosurfactant (BS) synthesis and their characterization. Out of 32 bacteria, the most promising isolate G6 was taken for a detailed study. Based on polyphasic taxonomy it was found to belong to the novelBacillussp.Nutrient screening for higher BS synthesis usingthe Plackett-Burman design(PBD) showed thatFeSO4, NaCl andwaste engineoil have a significant influence on BS production. Further optimization utilizingBox-Behnken design(BBD) revealed that the optimized condition for higher BS synthesis wasFeSO4(2.0 %), NaCl (2.0 %), and waste engine oil (2.2 %). The highest BS yield of 3.79 ± 0.265 g/L was attained with a cell biomass of 1.51 ± 0.234 g/L with 1.92 fold increase in the yield. Based on thin-layer chromatogram (TLC),Fourier transform infrared spectroscopy(FTIR), andGas Chromatogram-Mass Spectrometry(GC–MS) studies it was observed that the probable BS was a surfactin-like molecule that is stable at a wide range of temperature (up to 100° C), pH (5–9), and salinity (3–6 %).

Optimization of biosurfactant production by Pseudomonas aeruginosa using rice water and its competence in controlling Fusarium wilt of Abelmoschus esculentus

• Rice water was selected as the cheapest carbon source for formulating rice water medium in order to optimize biosurfactant synthesis by Pseudomonas aeruginosa . • The in vitro and in vivo antifungal efficacy and biocontrol strategy of the biosurfactant against Fusarium wilt of Abelmoschus esculentus were demonstrated. • The identification of the rhl gene, TLC, FT-IR, HPLC, and GC–MS analyses suggested the biosurfactant was rhamnolipid. • The biosurfactant that has been produced will aid in large-scale commercialization and will replace the usage of chemical surfactants in agricultural wastes. Biosurfactants, in comparison to chemical surfactants, have a greater impact on various applications due to their stability, degradability, and other physicochemical characteristics. Increasing yield and lowering production costs are critical to enhancing biosurfactant efficiency. Because biosurfactant production costs are determined by the least expensive substrate, agro-based industrial waste is one of the most promising economic strategies. Therefore, in this study we aimed to achieve the biosurfactant production in Pseudomonas aeruginosa by low cost rice medium using Response surface methodology (RSM) and assessed their competence in controlling the Fusarium wilt of Abelmoschus esculentus . Initially, the selected strain P. aeruginosa PBS29 was confirmed using 16S rRNA sequencing. Then, the 20% (v/v) of rice water was chosen as the cheapest carbon source for formulating rice water medium to optimize biosurfactant production using Central Composite Design. Enhanced biosurfactant yield of 9.35 g/l was attained through RSM by 0.59 fold higher than preliminary analysis. The model was significant with a regression coefficient of 0.98. The optimal condition was identified as 1.18% (w/v) nitrogen source [glutamic acid], pH 6.8, temperature at 37.4 °C, 2.5% (v/v) inoculum size, and 167.9 rpm agitation. In-vitro antifungal activity and biocontrol strategy of the biosurfactant demonstrated against Fusarium wilt of Abelmoschus esculentus at 100 μg/ml concentration by both soil drenching and foliar spray by pot trial. Furthermore, the detection of rhl gene, TLC, FT-IR, HPLC and GC–MS analyses suggested that the biosurfactant is rhamnolipid. Overall, in this study, the biosurfactant was synthesized using inexpensive rice water and substantiated its safe use as a biocontrol agent, which will help large-scale commercialization and interchange chemical surfactant used in agricultural wastes.

Optimization of Biosurfactant Production using Chrysene Degrading Bacteria Isolated from Marine Water

Biosurfactants were found to be a suitable candidate for environment clean-up and as an alternative to conventional treatment systems. In the present study, a polyaromatic hydrocarbon degrading organism Bacillus halotolerans was screened for its ability to produce biosurfactant during hydrocarbon degradation. The bio-surfactant activity of the organism was screened by using various methods like oil spreading technique, emulsification assay, foam height analysis and parafilm-M test. The design expert software was used to optimize the suitable conditions for the production of biosurfactants. The optimum conditions were determined as pH 6, Chrysene-100 ppm, nitrogen source -1000 ppm and at 144 hrs using the design of experiments. The organism showed good oil degradation capacity and maximum growth was achieved in 6 days. Biosurfactant obtained from the biodegradation medium was confirmed to be lipopeptide using TLC, FTIR and GCMS.

Stability and Optimization of Biosurfactant Production by Enterobacter Cloacae AYF1

Biosurfactants are diverse and heterogeneous group of microbial metabolites produced by certain bacteria and fungi that have both hydrophilic and hydrophobic components. This work was aimed at assessing the production, stability and optimization of rhamnolipid biosurfactant production conditions by Enterobacter cloacae AYF1 strain. Biosurfactant production, extraction, purification and optimization of the production conditions were assessed in mineral salt medium followed by stability tests under different ranges of pH, temperatures and salts concentrations using standard procedures. Gas chromatography mass spectroscopy (GC-MS) was used in characterizing the biosurfactant produced. One factor at a time (OFAT) optimization studies revealed that production of biosurfactant by Enterobacter cloacae AYF1 was optimal at 3% carbon source concentration (1.5mg/ml), pH 7.5 (1.7mg/ml), temperature 40ºC (1.8mg/ml) and incubation time of 96hrs (1.96mg/ml). The biosurfactant produced was characterized using gas chromatography mass spectroscopy (GC-MS). The results revealed the presence of different fatty acids at different peaks. This indicates that the production of rhamnolipid biosurfactant was optimal at the carbon concentration, pH, temperature and incubation time of 3%, 7.5, 40 ºC and 96hrs respectively. There is the need to use biotechnological approaches with a view to improving the performance of the bacterial strain.

A novel biosurfactant producing Kocuria rosea ABR6 as potential strain in oil sludge recovery and lubrication

Biosurfactants are amphiphilic molecules composed of a hydrophilic and hydrophobic moiety and had the ability to penetrate into different phases to reduce the surface tension. This features caused to oil recovery, lubrication and facilities of crude oil in pipeline. In current research Biosurfactant-producing strain was isolated from the storage tanks of the Isfahan Oil Refining Company in Iran, and screened by oil expansion test, droplet collapse, and surface tension reduction measurement. Hydrocarbon recovery from crude oil sludge was measured under constant conditions. The effect of factoring biosource lubrication on crude oil in pipelines was investigated in vitro. Also, the optimization of biosurfactant production in different conditions was measured as a single factor and using Response Surface Method (RSM). The best biosurfactant-producing bacterium was identified as Kocuria rosea ABR6, and its sequence was registered in the gene bank with access number of MK100469. Chemical analysis proved that the produced biosurfactant was a lipopeptide. 7% of crude oil was recovered from petroleum sludge by biosurfactant obtained from Kocuria rosea ABR6. Also, the speed of crude oil transfer in pipelines was upgraded as it could be said that for a certain distance the transfer time reduced from 64 to 35 s. The highest biosurfactant production was measured at pH 9, aeration rate of 120 rpm and 96 h after incubation. The use of biosurfactants produced by Kocuria rosea ABR6 is recommended to remove oil sludge and lubricate oil in pipelines recommended in the oil industry.

Optimization of biosurfactant production by marine Streptomyces youssoufiensis SNSAA03: A comparative study of RSM and ANN approach

The aim of the present study was to optimize media components to increase the production of biosurfactant (Bs) by Streptomyces youssoufiensis SNSAA03 employing response surface methodology (RSM) and artificial neural network (ANN). The RSM showed the following as the optimal condition for the production of Bs: glucose 15 g L–1, casein 0.3 g L–1, NaCl 2.25 g L–1 and pH 7. The superiority of the ANN model was proved with more accurate and consistent R2 values 0.998 when compared to RSM R2 value 0.980. The Bs produced by SNSAA03 was identified as glycolipid with 4–methoxy–2, 5–dimethylfuran–3(2H)–one and 1–Nanodecene acid. The micro titre plate assay showed that glycolipid at the concentration of 125 µg mL−1 was effective in controlling the biofilm formation by S. aureus. Thus, the glycolipid produced from SNSAA03 can be used as a potent antibiofilm agent.

Optimization of biosurfactant production from Pseudomonas sp. CQ2 and its application for remediation of heavy metal contaminated soil

The present study was conducted to enhance the biosurfactant production yield of Pseudomonas sp. CQ2 isolated from the Chongqing oilfield (China). Besides, the capability of biosurfactant and underlying mechanism for remediation of heavy metal contaminated soil was also investigated. Our results suggested that maximum biosurfactant production (40.7 g/L) was attained at 35 °C by using soybean oil and ammonium nitrate as carbon and nitrogen sources with pH 7, rotational speed of 175 rpm and inoculation ratio of 3%). The removal efficiencies of 78.7, 65.7 and 56.9% for Cd, Cu and Pb respectively were achieved at optimized bioleaching conditions (pH: 11, soil/solution ratio: 30:1 and non-sterilized soil), comparative tests between common chemical surfactants (SDS, Tween-80) and biosurfactants demonstrated the larger removal capacity of biosurfactants. Through SEM-EDX, it was found that the granular material disappeared, the content of Cd, Cu and Pb decreased significantly, and the soil surface became smooth with hole formation after soil washing following bioleaching. ATR-FTIR results showed that the carboxyl functional groups in biosurfactants could chelate heavy metals. These results indicated that biosurfactants from Pseudomonas sp. CQ2 could effectively eliminate Cd, Cu, and Pb from soil.

Nitrogen Optimization on Rhamnolipid Biosurfactant Production from Pseudoxanthomonas sp. G3 and Its Preservation Techniques

Biosurfactant is a microbial bioproduct that is used to reduce the surface tension, and can acted as emulsifier, dispersant, and anti-adhesive. Optimization of biosurfactant production needs to be done, not only to increase its production quantity, but also to reduce overall production cost. This study aims to determine the most suitable and optimum concentration of nitrogen source for biosurfactant production and its preservation techniques. The biosurfactant was produced by Pseudoxanthomonas sp. G3 using minimal salt medium with 2% light crude oil as carbon source and different nitrogen sources in the form of urea, sodium nitrate, and ammonium nitrate. The activity of biosurfactants were measured by emulsification index (E24), interfacial tense (IFT), oil drop assay, and dry weight. Potassium sorbate 0.2% (w/v) was used as preservative agent. The results showed that biosurfactant production using sodium nitrate as a nitrogen source provides the highest activity and yield. The E24 value was 76.63% and the clear zone diameter observed was 0.875 cm. The overall decreased in IFT was 35.4% and the biosurfactant dry weight was 0.45 gL-1. Microbial contamination occurred after 3 weeks of storage in the treatment without the addition of preservative. It also showed that the activity of biosurfactants (emulsification and IFT) were gradually decreased during storage. In conclusion, the optimum biosurfactant production by Pseudoxanthomonas sp. G3 was obtained by the addition of sodium nitrate 0.3% (w/v). Meanwhile, the most effective biosurfactant preservation method was by adding potassium sorbate which was stored at 4 ℃.

Optimization of biosurfactant production from chemically mutated strain of Bacillus subtilis using waste automobile oil as low-cost substrate

In this study, we have evaluated biosurfactant production potential of locally isolated strain of Bacillus subtilis (accession # KX788864) using waste automobile oil as cost-effective substrate with waste-to-value nexus. The wild strain of B. subtilis yielded 2.1 g/L biosurfactant, showed 42% emulsification index, and 38.27 cm2 of total oil displacement. The ethidium bromide (EtBr) mutagenesis was performed by exposing native B. subtilis strain with varying concentrations of EtBr (50, 100, 150 and 200 µg/10 mL) for different time intervals of 30, 60 and 90 min. Significantly higher yield (4.2 g/L) was attained from B. subtilis MS1 exposed to EtBr (50 µg/10 mL) for 60 min confirming the positive effect of mutagenesis. The mutated strain (B. subtilis MS1) showed improved emulsification index (50.53%) and oil displacement (49.32 cm2). Fourier-transform infrared (FTIR) spectroscopy analysis of biosurfactant produced from mutated strain (B. subtilis MS1) confirmed the lipopeptide nature of biosurfactant due to the existence of amide, amine and phospholipids functional groups. The maximum surfactant yielding mutant was proceeded for optimization using response surface methodology (RSM) technique and the interaction effect of different parameters was studied that allow biosurfactant hyperproduction. After optimization, maximum biosurfactant (4.7 g/L) was obtained at pH 7, temperature 35 °C, inoculum size 3 mL and an overall incubation of 120 h. In conclusion, the economical production of biosurfactant, along with novel functional properties (i.e., oil/hydrocarbon recovery and emulsification properties), this research may be valuable for numerous bioremediation and industrial biotechnological applications.

Application of foaming capacity as response value in optimization of biosurfactant production

Equipment for determination of surface tension (ST) which is the response value often used during biosurfactant optimization are not readily available in many laboratories. Foaming capacity (FC) was thus investigated as response value during biosurfactant optimization. Thirteen variations of glycerol-mineral salts medium were used to culture Pseudomonas sp. for biosurfactant production and optimization. Variations in the medium were based on pH, carbon-nitrogen (C-N) and carbon-phosphorus (C-P) ratio. Inoculated media were incubated at ambient temperature with orbital shaking at 150 rpm for four days. FC of the media were then determined and fitted using a polynomial model so as to obtain a prediction profile. The prediction profile was used to determine the combination of C-N ratio, C-P ratio, and pH that will lead to the highest FC. This combination was used in another experimental run for biosurfactant production, and at the end the culture was screened for biosurfactant activity. The results obtained showed that foaming was achieved in selected experimental runs, and the model for the prediction profile was worked out to be Y = -513.03 + 103.3804 X1 + 2.1211 X2 + 16.2848 X3 – 0.1108X1X2 – 0.3656 X1X3 – 0.0339X2X3 – 6.1730 X12 – 0.0099 X22 – 0.4432 X32. From the prediction profiles it was seen that the highest FC (32.04%) was achievable at combination of pH 7.0, C-N 40, and C-P 13. Biosurfactant activity of the culture with optimized combination showed that ST reduced from 56.43 to 35.28 mN.m-1. It is concluded that FC can be used in place of ST during biosurfactant optimization procedures.

Application of foaming capacity as response value in optimization of biosurfactant production

Application of foaming capacity as response value in optimization of biosurfactant production

Optimized Biosurfactant Production by <i>Pseudomonas aeruginosa</i> Strain CGA1 Using Agro-Industrial Waste as Sole Carbon Source

Biosurfactants are biomolecules produced by microorganisms, which possess several advantages over their chemical counterparts. Production can be cost-effective if renewable wastes are utilized as substrates. In this study, optimization of biosurfactant production by Pseudomonas aeruginosa strain CGA1 was carried out using response surface methodology. The conventional “One factor at a time” method of optimization was initially adopted to ascertain the impact of different renewable wastes on biosurfactant production. Four independent variables were tested: carbon and nitrogen concentration, medium volume, and inoculum size. Biosurfactant production was based on the emulsification index measurement. Results indicated that the preferred carbon source by the isolate was sugar cane molasses. A 2.31-fold increase in biosurfactant yield and emulsification index of 96.3% ± 0.75% under optimized cultural conditions of 20 g/L of molasses, 5 g/L of sodium nitrate, 1.93 ml inoculum size and 60 ml medium volume in 250 ml conical flask were obtained. The regression coefficient (R2) value of 84.15% implied adequate fitness of the model. The surface tension of distilled water was reduced from 72.1 mN/m to 35.0 ± 0.0 mN/m, and critical micelle concentration was attained at 60 mg·L-1. FTIR and GC-MS analysis indicated that the biosurfactant was a lipopeptide having characteristic lipid and peptide peak values. This study proves that the sole use of agro-industrial wastes for the production of biosurfactant is very efficient, and ensures the economic feasibility of biosurfactant production.

Optimization of bio-surfactant production by Azo-rhizobium strain isolated from oil-contaminated soil

Bio-surfactants are amphiphilic molecules possessing both hydrophilic and hydrophobic moieties. They are surface active agents that are produced extracellularly or as a part of the cell membrane by bacteria, yeast, and fungi. In the present study, the screening and optimization of bio-surfactant production were carried out using oil-contaminated soil sample. The cultural, morphological and biochemical tests as well as 16s rRNA analysis identified the most efficient bio-surfactant producer as Azorhizobium strain. The preliminary screening of bio-surfactant production was done with the help of oil displacement test, drop collapse test and observed hemolysis on superimposed blood agar plates. Optimization studies revealed that 2% inoculum size of test strain (1.0 O.D 530 nm) can be exploited for bio-surfactant production using 2% coconut oil and ammonium nitrate with a C: N ratio of 20:1 in MSM medium (pH 8.5) and incubation conditions of 30 ˚C for 96 h. The crude yield of bio-surfactant produced was estimated as 2.5 g/L. Further purification of crude bio-surfactant was carried out using acid hydrolysis and rotary vacuum evaporator. The bio-surfactant thus obtained successfully reduced the surface tension of the medium from 59 mN/m to 38 mN/m with E24 60%. The characterization studies of the purified bio-surfactant carried out by FTIR analysis confirmed it to be lipopolysaccharide type of bio-surfactant. Thus our current study suggests useful application of bio-surfactant producing bacterial strain that may be helpful in the petroleum industry for the purpose of recovery of petroleum and other oils from oily sludge, cleaning of oil storage tanks and bioremediation of oil-contaminated sites.

Optimization of bio-surfactant production by Azo-rhizobium strain isolated from oil-contaminated soil

Optimization of bio-surfactant production by Azo-rhizobium strain isolated from oil-contaminated soil

Optimization of biosurfactant production by Bacillus Subtilis N3-1P using the brewery waste as the carbon source

ABSTRACTBiosurfactants are biologically produced by microorganisms and therefore biodegradable, making ideal substitutes to chemical surfactants for various applications. Large scale production of biosurfactants is limited because of the high cost. The production cost could be reduced by optimizing cultural conditions and using wastes as substrates. In this work, the response surface methodology (RSM) was used to optimize biosurfactant production by Bacillus subtilis N3-1P strain using brewery waste as the sole carbon source. Five independent variables were varied; carbon and nitrogen concentration, agitation speed, temperature and initial pH. Surface tension and emulsification index were used to measure biosurfactant production. Results indicated that the ‘best’ surface tension and emulsification index were 27.31 mN m−1 and 63.11%, respectively, under optimized cultural conditions (7% (v v−1) brewery waste, 6.22 g L−1 ammonium nitrate, initial pH of 6.41, 150 rpm, and 27°C). The predicted responses were validated experimentally under the ‘optimum’ conditions, and 657 mg L−1 of biosurfactant was produced with a critical micelle concentration of 107 mg L−1.

Isolation, characterization, and optimization of biosurfactant production by an oil-degrading Acinetobacter junii B6 isolated from an Iranian oil excavation site

An enrichment program for hydrocarbon-utilizing bacteria capable of biosurfactant production isolated from soil samples was carried out on mineral salt medium supplemented with Iranian light crude oil (ILCO, 1%). The most promising isolate was identified as Acinetobacter junii B6 using 16S rDNA sequencing and biochemical characterization. Time course profile of biosurfactant production using A. junii B6 showed surface tension reduction in the culture broth to 45 mN/m after 48 h incubation and a 51% emulsification of ILCO. Scanning electron microscopy (SEM) analysis of A. junii B6 showed the tendency of bacterial cells to adhere to each other and to the oil droplets. Application of two-level fractional factorial design showed that surface tension of culture broth was maximally reduced to 38 mN/m in the presence of NaNO3 (2 g/L), ILCO (5%), temperature of 25 °C, aeration rate of 300 rpm, and inoculum size of 2%. GC-MS analysis of the culture broth showed the ability of A. junii B6 to degrade most of the alkanes' components of ILCO when used as sole carbon source. Both strain and product has potential applications in various oil industries including bioremediation and enhanced oil recovery.

Optimization of biosurfactant production from Vibrio sp. BSM-30 isolated in tropical waters

The strain BSM-30 (Vibrio sp.), isolated from Chinese tropical waters, could be a biosurfactant producing bacteria according with results obtained by the oil spreading method. The culture conditions for biosurfactant production were tested respectively such as inoculation (2%,6%,10%,14% as setting), shaking speed(120 r/min,150 r/min,180 r/min as setting), temperature (25°C,30°C,35°C as setting), pH (7,8,9 as setting), salinity (1.5%, 2.5%, 3.0%, 4.5%, 5.5% as setting), which results showed that the best culture conditions for BS production were 10% inoculation quantity, 180 r/min, 25°C, pH 8, and 3.5% salinity. The optimization of carbon sources (20g/ of glucose, 20g/L of starch, 20g/L of paraffin oil 20g/L of diesel, 20g/L of oil as setting) and nitrogen sources (6g/L of NaNO3,7.1g/L of KNO3,5.6g/L of NH4NO3,9.3g/L of (NH4)2SO4, 4.2g/L of CO(NH2)2 as setting) were also tested, which results showed that the best nitrogen source and carbon source were (NH4) 2SO4 and soluble starch.

Optimization of biosurfactant production by Bacillus brevis using response surface methodology

The present study aims to evaluate and validate a statistical model for maximizing biosurfactant productivity by Bacillus brevis using response surface methodology. In this respect, twenty bacterial isolates were screened for biosurfactant production using hemolytic activity, oil spreading technique, and emulsification index (E24). The most potent biosurfactant-producing bacterium (B. brevis) was used for construction of the statistical response surface model. The optimum conditions for biosurfactant production by B. brevis were: 33°C incubation temperature at pH 8 for 10 days incubation period and 8.5g/L glucose concentration as a sole carbon source. The produced biosurfactant (BS) (73%) exhibited foaming activity, thermal stability in the range 30–80°C for 30min., pH stability, from 4 to 9 and antimicrobial activity against (Escherichia coli). The BS gave a good potential application as an emulsifier.

Optimization of biosurfactant production in soybean oil by rhodococcus rhodochrous and its utilization in remediation of cadmium-contaminated solution

Biosurfactant production by Rhodococcus rhodochrous in soybean oil was developed, where the effect of medium composition and fermentation time were evaluated. The optimum condition for biosurfactant production was achieved when a medium containing 30 g/L TSB (tryptic soy broth) and 20% v/v soybean oil was used as media with 7 days of fermentation. Biosurfactant was identified as glycolipids type biosurfactant which had critical micelle concentration (CMC) value of 896 mg/L. The biosurfactant had oil in water emulsion type and was able to reduce the surface tension of palm oil about 52% which could stabilize the emulsion up to 12 days. The batch removal of cadmium metal ion by crude and partially purified biosurfactants have been examined from synthetic aqueous solution at pH 6. The results exhibited that the crude biosurfactant had a much better adsorption ability of Cd(II) than that of partially purified biosurfactant. However, it was found that there was no significant difference in the adsorption of Cd(II) with 5 and 10 minutes of contact time. The results indicated that the biosurfactant could be used in remediation of heavy metals from contaminated aqueous solution.

Optimization of biosurfactant production by Streptomyces isolated from Egyptian arid soil using Plackett–Burman design

Surfactants are amphipathic molecules which reduce surface tension, widely used in pharmaceutical, cosmetic and food industries. The current study aimed to isolate Streptomyces species from Egyptian arid soil, efficient in biosurfactant production. Fifty isolates of Streptomyces were obtained from arid soil in Egypt. Primary screening, using lipase, drop collapse, parafilm M and haemolysis tests, showed 37 isolates (74%) having biosurfactant activity. Secondary screening resulted in 17 isolates based on their activities using emulsification index method. Isolate 5S was selected as the most efficient in biosurfactant production, having emulsification index equal to 31.74%. Plackett–Burman design was adopted to evaluate the factors affecting biosurfactant production. Among the tested nine variables of starch nitrate medium, treated molasses, peptone, Tween 80, incubation period and inoculum size were identified as significant factors affecting biosurfactant production. Cultivation of isolate 5S in the optimized medium elevated biosurfactant production to 42.68% (according to emulsification index) from 31.74%, obtained in un-optimized medium.