Isolation Of Biosurfactant Producing Bacteria Research Articles

Isolation of Biosurfactant Producing Bacteria from Oil-Spilled Soil and Characterization of Their Secreted Biosurfactants in Pathogen-Inhibition and Oil-Emulsification

Isolation of Biosurfactant Producing Bacteria from Oil-Spilled Soil and Characterization of Their Secreted Biosurfactants in Pathogen-Inhibition and Oil-Emulsification

Isolation of Biosurfactant-Producing Bacteria and Their Co-Culture Application in Microbial Fuel Cell for Simultaneous Hydrocarbon Degradation and Power Generation

Biosurfactant-producing microorganisms improve the efficacy of hydrocarbon biodegradation as the biosurfactant is essential in making hydrocarbons available for breakdown. The present study reports the isolation of biosurfactant-producing bacteria that can be used for crude oil remediation and to characterize the biosurfactant generated during the breakdown of crude oil. This study also reports evaluating the synergism and potentiality of biosurfactant-producing bacteria for simultaneous hydrocarbon biodegradation and power generation. Two bacterial strains (Bacillus subtilis strain B1 and Pseudomonas aeruginosa strain B2) were isolated from petroleum-contaminated soils, which are found effective in producing biosurfactants and degrading crude oil as the sole carbon source. B. subtilis B1 exhibited a higher potential for biosurfactant production and crude oil degradation than P. aeruginosa B2. The FTIR and GC-MS analysis were conducted for further characterization of the biosurfactant, which revealed that the surfactant produced by strain B1 and B2 was surfactin and rhamnolipid, respectively. The application of the B1 and B2 co-culture in microbial fuel cells (MFCs) showed synergism among them and resulted in a maximum power density production of 6.3 W/m3 with an open circuit voltage of 970 mV while degrading 2.5% v/v crude oil containing anolyte. The findings indicate that the co-culture of isolated crude oil-degrading strains has great potential for enhanced power generation and the bioremediation of hydrocarbon-contaminated environments. Moreover, the synergism of isolated strains in MFCs suggested their potent applicability in environmental, energy, and industrial sectors as an economical and feasible alternative to the existing technologies.

Isolation of Biosurfactant-Producing Bacteria from Petroleum- Polluted Mangrove Swamp Samples Using Selective Enrichment Procedure

Fifty-five morphologically distinct isolates were obtained from crude oil polluted mangrove swamp samples collected from Niger-Delta, region of Nigeria, using three microbiological media for selective enrichment. Isolation from the cultivation media was performed at 3 intervals (Days 3, 7 and 14). 35% of filamentous/irregular shaped colonies were obtained at longer incubation (Day 14) and 80% of colonies were gram positive rods. Cell-free supernatants from twenty-two isolates indicated biosurfactant activity in one or all test media (Nutrient medium (NB), NB + 2% glycerol and Mineral salt medium + 2% glucose) based on results from five screening tests; surface tension, emulsification index, oil displacement, drop collapse and haemolysis. These isolates demonstrated medium-dependent biosurfactant synthesis with 16 isolates producing biosurfactant in NB + 2% glycerol compared to other test media. Results of surface tension reduction ranged from 29.8 to 48.6 mN m-1 but results did not correlate well with emulsification index, hence combination of screening methods was essential for confirmation of biosurfactant-producing isolates. 16SrRNA sequencing identified isolates belonging to Bacillus, Serratia and Paenibacillus genera. Among these isolates, SB14-1 identified as Bacillus humi utilised three test media showing ST reduction (34.1 to 48.6 mN m-1) and emulsification of paraffin oil (49.1 to 62.6%). To date, there are no reports of biosurfactant synthesis from Bacillus humi amongst other Bacillus species. The use of selective enrichment media and combination of biosurfactant screening tests could be a successful approach towards isolation of possibly new biosurfactant-producing strains and/or hyper-producers.

Identification and Isolation of Biosurfactant Producing Bacteria from Oil Spill Contaminated Soil of Ennore

The bacteria were isolated from the soil samples that were collected in oil spill-contaminated sites located at Kamarajar port in Ennore, Chennai, Tamil Nadu, India. The main aim of this study was to isolate the bacteria that produce biosurfactants from the oil spill contaminated soil in Ennore, Chennai and to produce a growth medium for the bacteria to enrich the growth of the bacteria. For the growth of bacteria, Minimal Medium (MM) and Mineral Salt Medium (MSM) were the two types of growth enrichment medium used and serial dilutions of the bacterial cultures were done for the production of colonies. The objective of the study was to perform tests to identify whether the bacteria can be able to produce the bio-surfactant or not. Tests like Drop collapse method and oil displacement method were done to check for the production of biosurfactant. The bacteria which produce the surface active substances were the one which produces the bio-surfactant. As a result, gram staining was done for identifying the nature of bacteria. Most of the gram positive bacteria are known to have the properties of bio-surfactant. These bacteria has peptidoglycan outer layer that are easier to kill - their thick peptidoglycan layer which absorbs antibiotics and cleaning products easily. Hereby we conclude that the bacteria which produces the biosurfactant is Bacillus and further studies should be done on identifying the bacteria at the species level. Therefore, this study was done to eradicate the oil spill contaminated areas with the help of bacteria in a natural way without harming the environment. Bacillus sp. is a gram-positive bacteria which helps in, liver and renal function, and act as an inflammatory markers and in maintaining blood glucose levels. It does not have an outer cell wall and only has a membrane of peptidoglycan which makes the Bacillus more absorbent and eco-friendly.

Isolation of biosurfactant producing bacteria from Potwar oil fields: Effect of non-fossil fuel based carbon sources

AbstractBiosurfactants are surface-active metabolites produced by microorganisms. Biosurfactants tend to solubilize hydrocarbons in the surrounding environment, by reducing surface tension and increasing carbon uptake. In this study, isolation of biosurfactant producing bacteria and effect of renewable, non-fossil fuel based carbon sources on production were assessed. Oil field produced water was collected from Potwar oil fields and thirteen strains were isolated. Strains were screened for biosurfactant production by hemolysis test, emulsification index test, emulsification assay, oil displacement test, drop collapse test, tilted glass slide test and oil coated agar emulsification test. Strains were further screened for maximum production on cost effective renewable carbon source e.g. glucose, waste glycerol and animal fat. Promising strains were identified as Bacillus subtilis (MH142143), Pseudomonas aeruginosa (MH142144), Bacillus tequilensis (MH142145) and Bacillus safensis (MH142146) by 16S rRNA gene sequencing. Among all isolates, highest biosurfactant production on glucose (37%), glycerol (48%) and animal fat oil (49%), after 24 h cultivation was by Pseudomonas aeruginosa. Biosurfactant showed similarity to rhamnolipids by Thin Layer Chromatography (TLC). Assessment of bioactive propertiaes of rhamnolipid showed strong antimicrobial activity against Bacillus spp. Future investigations can be focused on application of these strains in environmental as well as industrial sectors.

Isolation of Potential Biosurfactant Producer from Oil Contaminated Soil and Water

Biosurfactants are extracellular macromolecules produced by bacteria, yeast, and fungi when grown on different carbon sources. It has the ability to reduce the surface tension of a liquid, interfacial tensions between two liquids and between a liquid and a solid. This study was conducted to isolate potential biosurfactant producers from oil-contaminated soil and water. Soil and water samples were obtained from the food court area in front of Universiti Malaysia Kelantan, Jeli Campus. Isolation of biosurfactant producing bacteria was carried out on minimal salt medium (MSM) supplemented with palm oil as the sole carbon source. Five potential biosurfactant producers; WS2, WS4, WS5, SS2 and SS5 were successfully isolated and identified by 16S rRNA analysis. Isolate WS4, SS2 and SS5 showed the highest similarity to Klebsiella sp and the other two isolates, WS2 and SS5 showed the highest similarity to Pseudomonas sp. and Nanobacterium sp. respectively. While Klebsiella sp. and Pseudomonas sp. were reported as prevalent biosurfactant producer, no report is available on the production of biosurfactants by Nanobacterium sp. All isolates showed variation in biosurfactant characterization assays which are emulsification test, drop collapse test, oil spreading test, blood haemolysis and blue agar plate assay.

Isolation of Biosurfactant Producing Bacteria from Garbage Soil

Biosurfactants are amphiphilic compounds secreted extra-cellularly by microorganisms. Biosurfactants contains hydrophobic and hydrophilic moieties that reduce the surface tension and interfacial tension by accumulating at the interface of hydrophobic substances and thus increase the solubility, bioavailability and subsequent biodegradation of the hydrophobic or insoluble organic compounds. The present study focused on the isolation of biosurfactant producing microorganisms from garbage soil. Isolated microorganisms were identified based on the biochemical characterization as Staphylococcus aureus and Bacillus cereus. Qualitative analysis of biosurfactant production was identified by performing assays like hemolysis on blood agar, penetration assay and C-TAB agar assay. The present study confirms that isolated microorganisms have biosurfactant property.

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Rhamnolipids Produced by Indigenous Acinetobacter junii from Petroleum Reservoir and its Potential in Enhanced Oil Recovery.

Biosurfactant producers are crucial for incremental oil production in microbial enhanced oil recovery (MEOR) processes. The isolation of biosurfactant-producing bacteria from oil reservoirs is important because they are considered suitable for the extreme conditions of the reservoir. In this work, a novel biosurfactant-producing strain Acinetobacter junii BD was isolated from a reservoir to reduce surface tension and emulsify crude oil. The biosurfactants produced by the strain were purified and then identified via electrospray ionization-Fourier transform ion cyclotron resonance mass spectrometry (ESI FT-ICR-MS). The biosurfactants generated by the strain were concluded to be rhamnolipids, the dominant rhamnolipids were C26H48O9, C28H52O9, and C32H58O13. The optimal carbon source and nitrogen source for biomass and biosurfactant production were NaNO3 and soybean oil. The results showed that the content of acid components increased with the progress of crude oil biodegradation. A glass micromodel test demonstrated that the strain significantly increased oil recovery through interfacial tension reduction, wettability alteration and the mobility of microorganisms. In summary, the findings of this study indicate that the newly developed BD strain and its metabolites have great potential in MEOR.

Aislamiento de microorganismos productores de biosurfactantes y lipasas a partir de efluentes residuales de camales y suelos contaminados con hidrocarburos

Surfactants are amphipathic molecules which reduce stress at the interface, thereby increasing water solubility and availability of organic compounds are produced by bacteria, fungi, and yeasts. For the isolation of biosurfactant producing bacteria and lipases, was plant in inducing means 10% of sewage effluent from slaughterhouses and soils contaminated with hydrocarbons Province taps Trujillo - Peru. Isolates were seed in agar cultures lecithin and rhodamine agar for determination of lipase producers. The selected microorganisms were plate on blood agar and agar Siegmund and Wagner (SW) for preliminary selection of biosurfactants bacteria. Later it determined to those microorganisms producing surfactants by Emulsification Index (EI) and measurement of surface tension. Positive cultures these tests were select and identified by API galleries and molecular identification by DNA sequencing. Crops further reduced surface tension were P. aeruginosa and B. amyloliquefaciens, reaching reduce surface tension to 27.2 ± 0.4 and 31.03 ± 0.4 mN / m respectively, while P. stutzeri and Pseudomonas sp. reduced surface tension to 48.3 and 52.6 ± 0.4 mN / m respectively. We conclude, therefore, that 49 producing crops lipases province Trujillo, of which it was determined that Pseudomonas stutzeri, Pseudomonas sp, Pseudomonas aeruginosa and Bacillus amyloliquefaciens are producers of biosurfactants and can be applied in the area of biotechnology was obtained environmental for the removal and biodegradation of contaminants.

Evaluation of biosurfactant producing activity of Bacillus thuringiensis-VITNDUVNB04 isolated from oil contaminated soil by various methods

Biosurfactants are the surface active molecules that are produced by microorganisms. The present study focused on the isolation of biosurfactant producing bacteria from oil contaminated soil samples of an automobile mechanical workshop near VIT University, Vellore. A total of 5 bacterial colonies were isolated from the collected samples (DNT-1 to DNT-5). Among them the isolate DNT-2 showed maximum biosurfactant production followed by DNT-3 and DNT-5. In primary screening DNT-2 showed a good haemolytic activity with a zone of clearance of 18 mm followed by DNT-3 (12 mm) and DNT-5 (10 mm). All three isolates were positive for lipase production and capable of degrading tributyrin. In the secondary screening method, DNT-2 showed high activity with 22 mm of oil spreading activity and 20 mm of drop collapsing activity. The emulsifying capacity of DNT-3, DNT-2 and DNT-5 was evaluated using E24 emulsification index where isolates showed 52%, 86% and 43% of activity respectively. Among all the isolates DNT-2 showed high surfactant production in all screening assays. Potential isolate DNT-2 was identified as Bacillus thuringiensis using biochemical tests and molecular 16s r RNA sequencing analysis..

Isolation of Biosurfactant Producing Bacteria from Petroleum Contaminated Terrestrial Samples that Collected in Bangkok, Thailand

Biosurfactants are amphiphilic molecules with effective surface-active and biological properties applicable to replace synthetic surfactant in several industries and processes. In this study biosurfactant producing bacteria were isolated from 50 terrestrial samples collected in areas contaminated with petroleum in several districts in Bangkok, Thailand. The biosurfactant producing ability of bacteria was investigated by oil displacement agar plate and emulsification index method. A total of 42 biosurfactant producing bacteria were isolated from the samples. The best biosurfactant producing isolate that showed the highest oil displacement and emulsification activity was identified as Serratia marcescens based on microscopic and biochemical analysis.

Biosurfactant production by Arctic and Antarctic bacteria growing on hydrocarbons

The risk of hydrocarbon contamination in marine polar areas is constantly increasing. Autochthonous bacteria, due to their ability to cope and survive under extreme environmental conditions, can play a fundamental role in the hydrocarbon degradation. The degradation process is often enhanced by the production of biosurfactant molecules. The present study reports for the first time on the isolation of biosurfactant-producing bacteria from Arctic and Antarctic shoreline sediments. A total of 199 psychrotolerant bacterial isolates were obtained from hydrocarbon-amended (with crude or diesel oil) microcosms. A total of 18 isolates were selected for their ability to grow in the presence of crude oil and produce biosurfactants, as it was revealed by the production of good E24 values (≥50 %) and/or reduction in the surface tension (under 30 mN/m). The positive response of the isolates to both tests suggests a possible production of biosurfactants with emulsifying and interfacial activities. Biosurfactant-producing isolates were mainly affiliated to the genera Rhodococcus (14 isolates), followed by Pseudomonas (two isolates), Pseudoalteromonas (one isolate) and Idiomarina (one isolate). Thin-layer chromatography of biosurfactant crude extracts revealed that the majority of the selected isolates were able to produce glycolipidic surfactants. Our results enlarge the knowledge, which is still poor and fragmentary, on biosurfactant producers from polar areas and indicate marine polar sediments as a source of bacteria with potential applications in the remediation of hydrocarbon-contaminated cold environments.

Evaluation of Screening Methods for the Isolation of Biosurfactant Producing Marine Bacteria

Hemolytic assay, bacterial adhesion to hydrocarbons (BATH) assay, drop collapse assay, oil spreading assay, emulsification assay and surface tension measurement were compared for their reliability and ease of use to screen biosurfactant producing bacteria. 105 bacterial strains were screened for their biosurfactant production. Results from screening methods revealed that hemolytic and BATH assays are not reliable methods to check biosurfactant production because hemolytic assay relies on lysis of blood cells, which can be caused by compounds produced by microbes other than the biosurfactants. In that case hemolytic assay may include strains that do not produce biosurfactant. BATH assay relies on hydrophobicity or adherence of cells with hydrophobic compounds. Results obtained in BATH assay for three bacterial isolates, Branhamella catarrhalis, Citrobacter intermedius and Klebsiella ozaenae were positive, but when the same isolates screened for drop collapse, oil spreading, and surface tension measurement the results were negative. This indicates that these bacterial cells are either acting as biosurfactant themselves (cell surface) or they don’t have the ability to produce biosurfactants. In conclusion, drop collapse and oil spreading assays are reliable methods to screen large number of samples. However, for confirmation surface tension measurement can be used at the end. Another interesting finding in this study was the use of crude oil as hydrophobic substrate for the isolation of biosurfactant producing bacterial strains; this approach may be useful during the initial isolation of biosurfactant producing bacteria to reduce the number of strains to be screened.

Isolation of potent biosurfactant producing bacteria from oil spilled marine water and marine sediments

Biosurfactants produced by bacteria are surface-active compounds involved in the degradation of hydrocarbons. They are heterogeneous group of surface active molecules produced by microorganisms, which adhere to cell surface or are excreted extracellularly in the growth medium. These can be divided into low- molecular weight molecules, that lower surface and interfacial tensions efficiently and high-molecular-weight polymers that bind tightly to surfaces. These surfactants, produced by a wide variety of microorganisms, have very different chemical structures and surface properties. It is therefore reasonable to assume that different groups of biosurfactants have different natural roles in the growth of the producing microorganisms. In this study, isolation and identification of biosurfactant producing bacteria were assessed from oil-spilled seawater collected from harbors and docks from Arabian Sea (Mumbai), India. The potential application of these bacteria in microbial enhanced oil recovery (MEOR) was investigated. To confirm the ability of isolates in biosurfactant production, various biosurfactant activity assay tests and measurement of surface tension were conducted. Hemolysis was used as a criterion for the primary isolation of biosurfactant producing bacteria. Effects of different pH, salinity and temperature on biosurfactant production were also studied. Among all of the isolated strains, Pseudomonas sp. (MW2) showed high salt tolerance, successful production of biosurfactant in a vast pH and temperature domain and reduced surface tension of the medium to value below 40 mN/m. This strain is a potential candidate for MEOR. The MW2 biosurfactant component was detected as glycolipid in nature. Key words : Biosurfactant, emulsification, extreme conditions, microbial enhanced oil recovery, marine water.

Isolation of Biosurfactant Producing Bacteria From Poultry Breast Skin

Background: Biosurfactants are surface-active compounds produced by some microorganisms. Objectives: In this study, we collected surface skin samples from breast of poultry (chicken, turkey, and, quail) and screened for biosurfactant-producing bacteria. We also determined the genera of cultured strains. Materials and Methods: 33 hemolytic bacterial strains (15, 11, and 7 isolates from chicken, turkey, and quail, respectively) were isolated; oil spreading (OS) and bioemulsifying activities were measured for all isolates. Results: Two isolates of chicken (6.06%), three of turkey (9.1%), and three of quail (9.1%) were positive in all examinations (hemolysis, emulsification index (E24) and oil spreading). In total, eight isolates (24.24%) were positive in all examinations, out of them, seven isolates (87.5%) were gram positives, mainly belonged to Bacillus spp., Staphylococcus spp. and Lactobacillus spp. 31 isolates (93.9%) (out of 33 hemolytic isolates) were positive in oil spreading test while only eight isolates (24.24%) were positive in E24 test. Conclusions: The results showed that biosurfactant-producing bacteria are distributed in breast skin surface of examined birds. Further investigation about the composition of biosurfactants and phylogenetic determination of biosurfactant producing bacteria is suggested.

Isolation of Biosurfactant Producing Bacteria From Poultry Breast Skin

Biosurfactants are surface-active compounds produced by some microorganisms. In this study, we collected surface skin samples from breast of poultry (chicken, turkey, and, quail) and screened for biosurfactant-producing bacteria. We also determined the genera of cultured strains. 33 hemolytic bacterial strains (15, 11, and 7 isolates from chicken, turkey, and quail, respectively) were isolated; oil spreading (OS) and bioemulsifying activities were measured for all isolates. Two isolates of chicken (6.06%), three of turkey (9.1%), and three of quail (9.1%) were positive in all examinations (hemolysis, emulsification index (E24) and oil spreading). In total, eight isolates (24.24%) were positive in all examinations, out of them, seven isolates (87.5%) were gram positives, mainly belonged to Bacillus spp., Staphylococcus spp. and Lactobacillus spp. 31 isolates (93.9%) (out of 33 hemolytic isolates) were positive in oil spreading test while only eight isolates (24.24%) were positive in E24 test. The results showed that biosurfactant-producing bacteria are distributed in breast skin surface of examined birds. Further investigation about the composition of biosurfactants and phylogenetic determination of biosurfactant producing bacteria is suggested.

Isolation of biosurfactant-producing bacteria from the Rancho La Brea Tar Pits

This research was conducted to identify culturable surfactant-producing bacterial species that inhabit the 40,000-year-old natural asphalt seep at the Rancho La Brea Tar Pits in Los Angeles, CA. Using phenanthrene, monocyclic aromatic hydrocarbons, and tryptic soy broth as growth substrates, culturable bacteria from the tar pits yielded ten isolates, of which three species of gamma-proteobacteria produced biosurfactants that accumulated in spent culture medium. Partially purified biosurfactants produced by these strains lowered the surface tension of water from 70 to 35-55 mN/m and two of the biosurfactants produced 'dark halos' with the atomized oil assay, a phenomenon previously observed only with synthetic surfactants. Key findings include the isolation of culturable biosurfactant-producing bacteria that comprise a relatively small fraction of the petroleum-degrading community in the asphalt.

Isolation of Biosurfactant Producing Bacteria From Oily Skin Areas of Small Animals

: Background: Biosurfactants or microbial surfactants are surface-active biomolecules that are produced by a variety of microorganisms. They are a structurally diverse group of surface-active molecules and are highly sought after biomolecules for both present and future applications.Objectives: The aim of the present study was to isolate and identify biosurfactant producing bacteria from the ear canal and inguinal areas (oily skin areas) of dogs and cats.Materials and Methods: Eighty inguinal area and ear canal samples were collected from stray and owned dogs and cats (10 animals each, 20 samples) and screened for biosurfactant-producing bacteria using criteria such as hemolysis, oil spreading and E 24 emulsification index tests. The isolated strains were identified at genus level.Results: 42 hemolytic bacterial strains (20 from dogs and 22 from cats) were isolated. The owned animal’s samples had a higher population of positive strains than the stray ones. In total 11 isolates (26. 2%) were positive for all examinations, out of these 9 (21. 1%) isolates belonged to owned animals. 9 isolates (out of 11) (82%) were gram positive of which 4 (44. 4%) were Bacillus spp. and 3 (27. 2%) Lactobacillus spp.Conclusions: The results showed that biosurfactant producing bacteria are distributed in the oily skin areas of both dogs and cats. Further investigation into the composition of the biosurfactants and phylogenetic determination of biosurfactant producing bacteria is suggested. Implication for health policy/practice/research/medical education:Oily skin areas of small animals may be a source of novel biosurfactant-producing bacteria. Please cite this paper as:Ebrahimi A, Tashi N, Lotfalian S. Isolation of Biosurfactant Producing Bacteria From Oily Skin Areas of Small Animal. Jundishapour J Microbiol.2012.5(2):401-4. DOI: 10.5812/jjm.3190 Copyright © 2012 Kowsar Corp. All rights reserved.