Biosurfactant-producing Bacterial Strains Research Articles

Impact of biosurfactant produced by Bacillus spp. on biodegradation efficiency of crude oil and anthracene

Biosurfactants are surface active molecules generated by various microorganisms, including bacteria, actinobacteria, algae, and fungi. In this study, bacterial strains are isolated from soil contaminated with used motor oil and examined for potential biosurfactant production. A minimum salt medium (MSM), with crude oil as the only carbon source, is used to isolate potential biosurfactant-producing bacterial strains. About 23 strains are isolated, and all are subjected to the primary screening methods for biosurfactant production. Based on the emulsification index, oil displacement, and drop collapse screening methods, two isolates with potential biosurfactant-producing ability are selected for further studies. The synthesis of biosurfactants, crude oil and anthracene biodegradation is carried out with strains DTS1 and DTS2. Both strains show significant outcomes in crude oil degradation. In addition, both strains can utilize anthracene as the sole carbon source. During the degradation course, changes in the growth conditions are continuously monitored by measuring turbidity and pH. In this degradation study, the biosurfactant production aptitude of the isolated strains plays an essential role in increasing the bioavailability of hydrophobic hydrocarbons. These strains are identified down to the molecular level by employing 16S rRNA gene sequencing, and the acquired sequences are submitted to get the accession numbers. These prospective strains can be utilized to remediate hydrocarbon-contaminated environments.

Biodegradation of Waste Lubricant Oil by a Novel Isolated Biosurfactant Producer- Achromobacter xylosoxidans PSA5

Waste lubricant oil (WLO) is a critical environmental issue because of its recycle and disposal challenges. A potent biosurfactant-producing bacterial strain was isolated by the enrichment culture technique using WLO as the sole carbon source from the oil-contaminated soils. The strain was identified as Achromobacter xylosoxidans (NCBI GenBank accession number: MW673656) based on the morphological, biochemical, and 16S rRNA gene sequencing. The preliminary characterization of the produced biosurfactant by biochemical methods revealed its glycolipid nature. The strain possessed extensive biodegradative potential and degraded various components present in WLO. The gas chromatography-mass spectrometry (GC-MS) analysis showed that the strain could degrade hydrocarbons (C7-C22) and diethyl phthalate, a priority pollutant, with a relative degradation of 43.87% within 7 days of incubation. The biodegradation of WLO by the strain confers its use in the bioremediation of oil-polluted environments, and the simultaneous biosurfactant production is undoubtedly a step toward greener technology.

A biosurfactant-producing novel bacterial strain isolated from vermicompost having multiple plant growth-promoting traits.

The nutrient-rich vermicompost which is used as manure for the growth and development of plants is rich in microbial flora. These microbes protect the plants against several infectious pathogenic microbes. As certain microbes are known to produce biosurfactants as metabolites, an investigation was carried out to isolate biosurfactant-producing bacterial strains from vermicompost with the efficient antifungal property. From the study, it was revealed that biosurfactant-producing bacterial strains are present in the vermicompost. A total of nine bacterial strains were isolated from the vermicompost. Among them, one most efficient biosurfactant-producing bacterial strains with antifungal properties have been screened. After molecular characterization of the isolated strain, it was revealed that the bacterial strain is Bacillus licheniformis strain SCV1. The strain produces 3.4 ± 0.1 g/L of crude biosurfactant, which when column purified yields 3.1 ± 0.1 g/L of biosurfactant. The biosurfactant exhibited excellent emulsifying activity (E24 ) of 96.56% against crude oil. The produced biosurfactant was identified as a lipopeptide consisting of a mixer of surfactin and iturin. Furthermore, the biosurfactant exhibited significant antifungal activity against a wide range of phytopathogens, showing 76.3% inhibition against Sclerotinia sclerotiorum, 53% inhibition against Colletotrichum gloeosporioides, 51% against Fusarium verticillioides, and 36% against Corynespora cassicolla. Along with antifungal activities, the stain was found to exhibit multiple plant growth-promoting traits. This study, thus indicates that vermicompost might contain biosurfactant-producing microbes which can render protection to the plant against various phytopathogens by the production of biosurfactants and can also stimulate plant growth.

Enhanced Petroleum Removal by Potent Biosurfactant Producer Bacillus Subtilis CC9 Strain Isolated from an Oil Field

This study aimed to enhance the removal of petroleum with potent biosurfactant producing bacterial strains (Bacillius subtilis CC9, Enterobacter asburiae WMB1, Klebsiella oxytoca Rizhao 536-1, and Acinetobacter pittii PgBE252) isolated from an oil field. The biosurfactant production was determined by different screening methods such as hemolytic activity, drop-collapse, oil spreading, BATH, emulsification index and surface tension. While the presence of biosurfactant was detected with individual bacterial strain, B. subtilis CC9 strain was found to be quite effective on petroleum degradation (92%) with a high yield of biosurfactant production (0.95 ±0.05 g L-1). The degradation rate constant and half-life period of B. subtilis CC9 strain were calculated as K = 2.533 day−1, t1/2 = 0.273. It has been clearly shown with GC/MS analysis that B. subtilis CC9 strain was effective on the removal of long chain n-alkanes (C31 - C35) as well as short and medium chains. The FT-IR analysis also approved the presence of aliphatic and peptide moieties of biosurfactant extracted from B. subtilis CC9 strain. It is remarkably pointed out with this study that high yield of biosurfactant producing B. subtilis CC9 strain can be used in advanced petroleum bioremediation studies as an alternative to chemical surfactants.

Characterization of biosurfactant-producing bacterial strains isolated from agro-industrial wastes in southwestern, Nigeria

Introduction. The difficulty of managing trash and cleaning up the environment prompted interest in biosurfactants and surface-active proteins made by microbes. The study aims to augment bacterial isolates from agro-industrial wastes targeted for possible mass production of biosurfactants. Methods. Six agro-industrial wastes from Cassava, Palm kernel, and Sawdust from six agro-industrial sites within Ijebu area in Ogun State were collected for standard laboratory analyses in the Biotechnology Unit of the Federal Industrial Institute for Research, Oshodi (FIIRO). Five screening methods; blood hemolysis, lipase activity, blue agar hydrolysis, oil spreading, and emulsification index (EI24) were carried out to confirm biosurfactant production. Isolates with the highest hyper-production were subjected to 16rRNA molecular identification. Results. The study justified efficient biosurfactant production from 4 bacterial isolates out of 26 screened bacterial isolates from hydrocarbon degraders and 29 heterotrophic screened bacterial isolates, making a total of 55 screened bacterial isolates. Screening results reveal the emulsification capacities of identified Pseudomonas putida strain SG1, Acinetobacter baumanii strain MS14413, Bacillus zhangzhouensis strain cdsV18, and Burkholderia cepacia strain 717. Conclusion. Biosurfactant bacteria produced in all agricultural and industrial wastes considered in this study are capable of mass production.

Extracción y caracterización de un biosurfactante sintetizado por Acinotebacter baumannii

Biosurfactants are metabolites produced by microorganisms during their growth and reproduction. They are amphiphilic molecules capable of modifying surface and interfacial tension. A petrophilic, nitrogen-fixing, phosphorus-solubilizing, and biosurfactant-producing bacterial strain was isolated and characterized. The isolated strain was cultivated in Kim medium with a C:N ratio of 2:1, from which its growth kinetics was determined. Samples of the culture broth were taken at different times and the collapsed drop, oil dispersion and emulsification index tests were performed. Maximum culture growth occurred at 72 h. The biosurfactant was extracted from the crude cell-free broth using a mixture of solvents and a yield of 2.21 g/L was obtained. Thin-layer chromatography and FT-IR analyzes revealed the presence of a lipopeptide biosurfactant. Tensiometry tests showed that the biosurfactant was able to lower the surface tension of water from 72 mN/m to 40 mN/m and the critical micelle concentration was 0.058 (w/w). Probit analysis for toxicity determination showed that there was no weight loss in Eisenia foetida specimens in a range between 10000 and 50000 ppm.

Antifungal Activity of Rhamnolipid Biosurfactant Produced by Pseudomonas aeruginosa A4 against Plant Pathogenic Fungi

Biosurfactants have been shown to have a variety of other agricultural applications. Antimicrobial activity is a desirable property in a variety of biosurfactants. Several biosurfactants produced from bacteria have antibacterial activity against plant diseases, making them a promising biocontrol molecule for ensuring agricultural sustainability in the long run. The purpose of this research was to characterize biosurfactant produced at different carbon source by biosurfactant-producing bacterial strains isolated from contaminated soapstock sediment, as well as its antifungal properties. Biosurfactant-producing bacteria were isolated from soapstock-contaminated soil which bacteria were selected by using drop collapse and oil displacement tests. Twelve isolates reduced surface tension of culture broth from 50 to 31 - 43 mN/m. According to 16S rRNA sequence analysis, these isolates belong to 8 different genera (Acinetobacter, Citrobacter, Enterobacter, Klebsiella, Pantoea, Pseudomonas, Stenotrophomonas and Xanthomonas). The supernatant of Pseudomonas aeruginosa A4 grown in MSM supplemented with soapstock was the most effective biosurfactant against Aspergillus flavus F2, Aspergillus niger F14, Cunninghamella bertholletiae F1 and Rhizopus oryzae F5, inhibiting mycelium growth by 54, 61 59 and 50 %, respectively. The extract substance inhibited spore germination against A. flavus F2 and R. oryzae F5 with a minimum inhibitory concentration of 2.75 mg/mL. TLC, FT-IR, ESI-MS and GC-MS analysis demonstrated that the mono-rhamnolipids and di-rhamnolipids had the same 3-hydroxy fatty acid composition of C8, C10 and C12. Overall, the biosurfactant-producing strains identified in this investigation show promise for future development and application in cost-effective industrial-scale biotechnological processes.
 HIGHLIGHTS
 
 Biosurfactant producing bacteria were isolated from soapstock contaminated soil
 The supernatant of Pseudomonas aeruginosa A4 grown in MSM supplemented with soapstock was the most effective biosurfactant against plant pathogenic fungi
 Biosurfactant from Pseudomonas aeruginosa A4 was identified as rhamnolipid comprising of both mono and di-rhamnolipid congeners
 
 GRAPHICAL ABSTRACT

Screening of Biosurfactant Producing Bacteria Isolated from Hydrocarbon Contaminated Site and Their Potential in Biosorption of Pb(II) and Oil Biodegradation

Abstract In the present study, three potentially Pb(II)-resistant and biosurfactant-producing bacterial strains were isolated from a total of 23 strains using various screening methods, investigated for their biosorption of Pb(II) and used for the biodegradation of used motor oil. The results show that strain E1 (Bacillus haynesii) has significantly high efficiency in biodegradation of used motor oil, up to 82 % in the first three days. Maximum Pb(II) biosorption capacities of 238.09 mg/g and 99.01 mg/g were determined for strains E1 and F5 (Pseudomonas aeruginosa), respectively. The biosorption process was found to be in good agreement with the Langmuir isotherm for both E1 (R2 = 0.9614) and F5 (R2 = 0.9646), suggesting monolayer biosorption. The four common screening methods, namely the haemolytic assay, the determination of surface tension, the emulsifying activity and the foam test, were also correlated with the Pearson correlation method.

Mesocosm-based simulations to optimize a bioremediation strategy for the effective restoration of wildfire-impacted soils contaminated with high-molecular-weight hydrocarbons.

We obtained four microbial isolates from soil exposed to forest fire and evaluated their potential bioremediation activity when combined with a biosurfactant-producing bacterial strain for the decontamination of wildfire-impacted soil polluted with high-molecular-weight (HMW) hydrocarbons. We established mesocosm trials to compare three bioremediation strategies: natural attenuation, bioaugmentation and biostimulation. Chemical analysis, culture-dependent and culture-independent methods were used to evaluate the bioremediation efficiency and speciation of the microbial cenoses based on these approaches. After treatment for 90days, bioaugmentation removed 75·2-75·9% of the HMW hydrocarbons, biostimulation removed 63·2-69·5% and natural attenuation removed ~22·5%. Hydrocarbon degradation was significantly enhanced in the mesocosm supplemented with the biosurfactant-producing bacterial strain after 20 and 50days of treatment compared to the other bioremediation strategies. We found that the bioaugmentation approach was more effective than biostimulation and natural attenuation for the removal of HMW hydrocarbons from fire-impacted soil. Our study showed that micro-organisms from wildfire-impacted soil show significant potential for bioremediation, and that biosurfactant-producing bacterial strains can be combined with them as part of an effective bioremediation strategy.

Production of Bioemulsifiers from Bacteria Grown on Hydrocarbons

This work aimed to identify biosurfactant-producing bacterial strains isolated from hydrocarbon-contaminatedsites and to evaluate their biosurfactant properties. The drop-collapse method and minimal agar added witha layer of diesel as carbon source were used for screening, positive strains were able to grow in a liquidmedium, and surface tension and emulsification index were determined in cell-free supernatant and cellsuspension. 250 bacterial isolate were tested, and 16 were positive for the drop-collapse and hydrocarbonlayer agar method. Most of the strains were Pseudomonas, except for three strains (Acinetobacter, Bacillus,and Rhodococcus). Surface tension was similar in cell-free and cell suspension measurements, with valuesin the range of 58 to 26 (mN/m), and all formed stable emulsions with engine oil (77-92% E24). Consideringthe variety of molecular structures among microbial biosurfactants, they have different chemical propertiesthat can be exploited commercially, for applications as diverse as bioremediation or degradable detergents.

Isolation and Identification of Biosurfactant Producing Bacterial Strain from Saline Soil Samples in Iran; Evaluation of Factors on Biosurfactant Production

Background: Biosurfactants are a group of valuable amphiphilic molecules that have been widely used in different industries such as pharmaceuticals and cosmetics. Objectives: The current study aimed to screen and identify a bacterial strain able to produce biosurfactant from environmental soil samples, followed by evaluating factors that contribute to biosurfactant production. Methods: The biosurfactants production was evaluated every 24 hours by emulsification index (EI), oil spreading ability, and surface tension measurement. Thereafter, the sequences of 16S rDNA genes and biochemical tests were performed to identify the isolated bacterial. Furthermore, the effect of different factors, including nitrogen, vegetable oil, and cations on biosurfactant productivity of the selected bacterial strain, was also evaluated. Results: In primary screening, out of the six bacterial strains, only one isolate was found to produce a hemolytic zone on the blood agar plate. The EI index was increased in a time-dependent manner with a maximum EI of 10% after 12h. Surface tension measurement revealed decreased surface tension (51 mN/m) after 12 hours' incubation compared to that of the blank sample. The selected isolate was recognized as Bacillus atrophaeus by 16S rDNA gene sequencing and biochemical test. All applied factors in culture media could decrease the surface tension of culture broth compared to that of the blank sample. Conclusions: This study demonstrated a Bacillus strain able to produce biosurfactants. Such biosurfactant producers would have great potentials in bioremediation activities and microbial enhanced oil recovery.

Enhanced Production of Biosurfactant from Bacillus subtilis Strain Al-Dhabi-130 under Solid-State Fermentation Using Date Molasses from Saudi Arabia for Bioremediation of Crude-Oil-Contaminated Soils.

Crude oil and its derivatives are the most important pollutants in natural environments. Bioremediation of crude oil using bacteria has emerged as a green cleanup approach in recent years. In this study, biosurfactant-producing Bacillus subtilis strain Al-Dhabi-130 was isolated from the marine soil sediment. This organism was cultured in solid-state fermentation using agro-residues to produce cost-effective biosurfactants for the bioremediation of crude-oil contaminated environments. Date molasses improved biosurfactant production and were used for further optimization studies. The traditional “one-variable-at-a-time approach”, “two-level full factorial designs”, and a response surface methodology were used to optimize the concentrations of date molasses and nutrient supplements for surfactant production. The optimum bioprocess conditions were 79.3% (v/w) moisture, 34 h incubation period, and 8.3% (v/v) glucose in date molasses. To validate the quadratic model, the production of biosurfactant was performed in triplicate experiments, with yields of 74 mg/g substrate. These findings support the applications of date molasses for the production of biosurfactants by B. subtilis strain Al-Dhabi-130. Analytical experiments revealed that the bacterial strain degraded various aromatic hydrocarbons and n-alkanes within two weeks of culture with 1% crude oil. The crude biosurfactant produced by the B. subtilis strain Al-Dhabi-130 desorbed 89% of applied crude oil from the soil sample. To conclude, biosurfactant-producing bacterial strains can increase emulsification of crude oil and support the degradation of crude oil.

Antibiofilm Activity of Biosurfactant Produced by Bacterial Strain Isolated from Petroleum-Contaminated Sites

The escalating emergences of drug-resistant strains of Pseudomonas aeruginosa coupled with the decline in the development of new antibiotics have increased the morbidity and mortality rates during the past decades. The aim of the present work was to investigate the antibacterial and antibiofilm activity of biosurfactant produced by bacterial strain IHD19 isolated from hydrocarbon-polluted soil against biofilm forming clinical isolates of Pseudomonas aeruginosa. Isolates from petroleum-contaminated soil sample were screened and evaluated for biosurfactant production. Rapid methods such as drop collapse test, emulsification test, hemolysis test, lipase test were used for screening of biosurfactant production by bacterial strain IHD19. Bacterial strain IHD19 was selected on the basis of its efficiency to produce biosurfactant. Growth study of bacterial strain IHD19 was conducted at different physicochemical conditions aiming to find out the optimal condition for growth of bacterial strain IHD19 to obtain higher biosurfactant production. Thin-layer chromatography and FTIR analyses were done to characterize the biosurfactant produced by bacterial strain IHD19. Subsequently, the antibacterial activity of biosurfactant IHD19 on multidrug-resistant clinical isolates of Pseudomonas aeruginosa was investigated by agar well-diffusion method. Biofilm-forming clinical isolates of Pseudomonas aeruginosa were identified using crystal violet assay. Finally, the antibiofilm activity of biosurfactant IHD 19 was determined using microtiter plate assay and MBIC50 and MBIC90 were calculated. Bacterial strain IHD 19 was found positive for drop collapse test, lipase test and showed β-hemolysis on blood agar. The emulsification efficiency of biosurfactant IHD19 after 24 h (E24%) was 87.5% against engine oil. The thin-layer chromatography and FTIR analysis revealed the polymeric nature of the biosurfactant. Biosurfactant IHD19 displayed significant antibacterial and antibiofilm activity against twenty-five multidrug-resistant clinical isolates of Pseudomonas aeruginosa with more than 50% (MBIC50) in concentration range of 0.5–4 µg/ml and 90% biofilms inhibition (MBIC90) in concentration range of 16–32 µg/ml, respectively. Phylogenetic tree showed biosurfactant producing bacterial strain IHD19 to be relative to Bacillus family. With 16 s rRNA analysis the isolated bacterial strain IHD19 was identified as Bacillus pumilus. The current study proposes potential exploration of biosurfactant to subdue the issue of biofilms formation by MDR Pseudomonas aeruginosa.

Machine Learning Modeling of Aerobic Biodegradation for Azo Dyes and Hexavalent Chromium

The present study emphasizes the efficacy of a biosurfactant-producing bacterial strain Klebsiella sp. KOD36 in biodegradation of azo dyes and hexavalent chromium individually and in a simultaneous system. The bacterial strain has exhibited a considerable potential for biodegradation of chromium and azo dyes in single and combination systems (maximum 97%, 94% in an individual and combined system, respectively). Simultaneous aerobic biodegradation of azo dyes and hexavalent chromium (SBAHC) was modeled using machine learning programming, which includes gene expression programming, random forest, support vector regression, and support vector regression-fruit fly optimization algorithm. The correlation coefficient includes the dispersion index, and the Willmott agreement index was employed as statistical metrics to assess the performance of each model separately. In addition, the Taylor diagram was used to further investigate the methods used. The findings of the present study were that the support vector regression-fruitfly optimization algorithm (SVR-FOA) with correlation coefficient (CC) of 0.644, (scattered index) SI of 0.374, and (Willmott’s index of agreement) WI of 0.607 performed better than the autonomous support vector regression (SVR), gene expression programming (GEP), and random forest (RF) methods. In addition, the standalone SVR model with CC of 0.146, SI of 0.473, and WI of 0.408 ranked the second best. In summary, the SBAHC can be accurately estimated using the hybrid SVR-FOA method. In other words, FOA has proven to be a powerful optimization algorithm for increasing the accuracy of the SVR method.

Biochemical and Genomic Characterization of the Cypermethrin-Degrading and Biosurfactant-Producing Bacterial Strains Isolated from Marine Sediments of the Chilean Northern Patagonia.

Pesticides cause severe environmental damage to marine ecosystems. In the last ten years, cypermethrin has been extensively used as an antiparasitic pesticide in the salmon farming industry located in Northern Patagonia. The objective of this study was the biochemical and genomic characterization of cypermethrin-degrading and biosurfactant-producing bacterial strains isolated from cypermethrin-contaminated marine sediment samples collected in southern Chile (MS). Eleven strains were isolated by cypermethrin enrichment culture techniques and were identified by 16S rDNA gene sequencing analyses. The highest growth rate on cypermethrin was observed in four isolates (MS13, MS15a, MS16, and MS19) that also exhibited high levels of biosurfactant production. Genome sequence analyses of these isolates revealed the presence of genes encoding components of bacterial secondary metabolism, and the enzymes esterase, pyrethroid hydrolase, and laccase, which have been associated with different biodegradation pathways of cypermethrin. These novel cypermethrin-degrading and biosurfactant-producing bacterial isolates have a biotechnological potential for biodegradation of cypermethrin-contaminated marine sediments, and their genomes contribute to the understanding of microbial lifestyles in these extreme environments.

Bioremediation of 2,4-Diaminotoluene in Aqueous Solution Enhanced by Lipopeptide Biosurfactant Production from Bacterial Strains

AbstractThis research concentrated on the isolation and preparation of biosurfactant-producing bacterial strains for the degradation of aromatic amine compound 2,4-diaminotoluene (2,4-DAT). Three s...

Biodegradation of Crude Oil Through Biosurfactant Producing Bacterial Strains Isolated from Oily Sludge

Biodegradation of Crude Oil Through Biosurfactant Producing Bacterial Strains Isolated from Oily Sludge

Properties and potential application of efficient biosurfactant produced by Pseudomonas sp. KZ1 strain

Increasing use of biosurfactants has stimulated the search for new and efficient biosurfactant-producing bacterial strains, preferably nonpathogenic ones. The aim of the present study was to characterize a new isolated Pseudomonas sp. KZ1 strain and its exocellular surface active compounds. After examining several mineral media of different compositions, the bioreactor-scale production of biosurfactants under optimum conditions was tested. Then, the composition of the isolated biosurfactants was investigated by Fourier-transform infrared spectroscopy and gas chromatography-mass spectrometry analysis and their surface active properties were characterized by adsorption parameters. The results indicated that the Pseudomonas sp. KZ1 biosurfactant had the critical micelle concentration of 0.12 g L−1 and decreased the surface tension decreased to 31.7 mN m−1. Moreover, the biosurfactant increased the rate of biodegradation of diesel oil by the strains: Pseudomonas sp. KZ1, Pseudomonas sp. OS4 and Achromobacter sp. KW1. The obtained biosurfactant showing attractive properties is a promising and much ‘greener’ alternative in the application for surfactant-enhanced biodegradation of hydrocarbons.

A combination of rapid and easy assays of biosurfactant producing bacterial strain isolated from automobiles repairing workshop in Aligarh

Routine washing, cleaning, repairing, maintenance of cars, bikes, scooters and disposal of waste of all kinds are carried out in automobile workshops are common observations in Aligarh. Considering the likelihood of existence of biosurfactant producing organisms at hydrocarbon contaminated site, a large number of soil samples were collected and isolation was carried out. A total of ten bacterial strains ALIG (01–10) were isolated out of which only isolate АLIG01 grown on GSP agar, Maconkey agar as well as on Pseudomonas agar plates which indicated suspectedPseudomonas spp. and exhibits positive biosurfactant activity through penetration assay, oil spreading technique, beta hemolytic activity and ЕI24 (96%), positive blue plate agar plate (> 2сm), qualatitative analysis, tolerance against hydrocarbonm-xylene, and microplate assay. This isolate АLIG01 is a valuable source to investigate further for future agriculture plant pathology and industrial applications.

Screening, isolation and characterization of biosurfactant-producing Bacillus tequilensis strain ANSKLAB04 from brackish river water

Biosurfactants are amphiphilic compounds produced by micro-organisms that are excreted extracellularly and contain hydrophobic as well as hydrophilic moieties which reduce surface tension. These extracellular membrane vesicles aid in the uptake of organic molecules that are used as a source of energy for growth of micro-organisms. These micro-organisms have the ability to grow and proliferate on the oil-based medium like petro-contaminated soil and water. Oil spills and discharge from industry contaminates the water bodies and surrounding land. Many chemical-based technologies and manual cleaning are involved to reduce water and land pollution. Chemicals used are the surfactants that lead to the generation of toxic by-products. Hence, considerable attention has been given to surface-active molecules of biological origin called biosurfactants, produced by microbial species in and around the contaminated area. The present study focuses on identifying a novel biosurfactant-producing bacterial strain from brackish water. The bacterial consortium was isolated from targeted brackish water site. Each bacterial species was tested for its biosurfactant production ability using various assays. Haemolysis, oil spreading, cetyltrimethyl ammonium bromide agar plate, drop collapse, hydrocarbon overlay agar, hydrocarbon assay and lipase production were a few qualitative methods performed. Further, the most efficient biosurfactant producer was identified by measuring emulsification index, quantifying biosurfactant production and efficient reduction in surface tension. 16S rRNA gene sequencing was performed for phenotypic characterization of the biosurfactant-producing strain, using Sanger dideoxy sequencing method followed by the phylogenetic assessment. The isolate was found to be a novel Bacillus tequilensis strain, named as ANSKLAB04. The novel isolate was deposited in GenBank with Accession number KU529483.