Endophytic Bacillus amyloliquefaciens surfactin possesses anti-inflammatory potential through acetylcholinesterase and lipoxygenase inhibitory activities

Aims@#This study was aimed to investigate the anti-inflammatory and anti-rheumatoid effects of the Bacillus amyloliquefaciens derived surfactin.@*Methodology and results@#Crude and biosurfactant extracts were analyzed using thin-layer chromatography to determine the presence of biosurfactant. Both extracts were evaluated for their inhibitory effects against the acetylcholinesterase and 5-lipoxygenase enzymes. Human synovial cells were induced with TNF-α and IL-1β. The percentages of the cell viability for both normal and induced cells were determined with an MTT assay. Results showed that surfactin was detected in the biosurfactant extract and demonstrated higher inhibitory effects compared to the crude extract against both inhibitory enzymes acetylcholinesterse (IC50=30.60 μg/mL) and lipoxygenase (IC50=110.10 μg/mL). Both crudes showed no cytotoxic effects at the highest concentration used (50 μg/mL) against normal human synovial cells but showed active reactions against the induced cells. The anti-proliferative effects of biosurfactant and crude extracts were in dose-dependent manner.@*Conclusion, significance and impact of study@#Notably, surfactin obtained from B. amyloliquefaciens has shown an inhibitory effect against pro-inflammatory enzymes and cell viability of the induced rheumatoid arthritis cell line. These results highlighted the therapeutic potential of surfactin application as an anti-inflammatory agent for arthritis treatment. Further study is needed to elucidate the mechanisms underlying the anti-inflammatory effect of surfactin.

Comparative Roles of HBB5 Biosurfactant and Poultry Wastes in Polyaromatic Hydrocarbon Biodegradiation of Crude Oil-contaminated Sediment

The comparative study of poultry wastes- and HBB5 biosurfactant-mediated polyaromatic hydrocarbon biodegradation in sediment polluted with crude oil were investigated. The experiments were carried out for a period of 28 days by monitoring pH, nitrate, phosphate, polycyclic aromatic hydrocarbon and microbiological parameters using standard procedures. The pH values obtained ranged between 6.21 and 6.93 in days 1 and 28 for the most effective treatment recipes. Generally, there was depletion in the concentrations of nitrate and phosphate for all set ups, but the most effective recipe witnessed highest reduction. For the polycyclic aromatic hydrocarbons, the recipe with highest limiting nutrients depletion also recorded the most hydrocarbon loss, and yet highest increase in density of hydrocarbonoclastic bacteria and fungi. The sample containing polluted sediment + poultry wastes + HBB5 biosurfactant recorded PAH values of 1932.6472ppm on day 1 and 481.2272ppm on day 28. Total hydrocarbon-utilizing bacterial counts ranged from 1.48×104 cfu/g to 9.70×106 cfu/g, while hydrocarbon-utilizing fungal counts ranged between 2.30×103 cfu/g and 3.90×105 cfu/g. From the results obtained, poultry wastes combined with HBB5 biosurfactant recorded the highest efficiency in the biodegradation of polycyclic aromatic hydrocarbons in sediments, and HBB5 biosurfactant in isolation recorded higher degradation efficiency for polyaromatic hydrocarbons than the degradation effect mediated by poultry wastes alone. It is therefore recommended that a combination of surface-active agent, nutrient amendment source and viable microbial biomass be adopted and employed as potent recipe for the degradation of polyaromatic hydrocarbons in crude oil-contaminated sediments.

Field evaluation of biosurfactants, surfactin and di-rhamnolipid produced by Bacillus subtilis subsp. subtilis (VCRC B471) and Pseudomonas fluorescens (VCRC B426) against immature stages of the urban malaria vector Anopheles stephensi

Background & objectives: Bacillus subtilis subsp. subtilis (VCRC B471) and Pseudomonas fluorescens (B426) produce mosquitocidal biosurfactant, surfactin and di-rhamnolipid. The objective of the study was to carry out a small-scale field evaluation of the two biosurfactants to determine the efficacy, application dosage, residual activity and frequency of application against Anopheles stephensi immatures in selected sites in Goa, India. Methods: Surfactin (VCRC B471) and di-rhamnolipid (VCRC B426) were formulated as aqueous suspensions (5% AS), and were applied at the dosages of 34, 51 and 68 mL/m2 and 27, 41 and 54 mL/m2 respectively. Two experiments were carried out with the two formulations. Results: Surfactin (VCRC B471) formulation was effective at all the dosages and there was sustained reduction (>80%) in immature density in the treated sites up to 18 days in experiment 1 and up to 15 days in experiment 2. No pupae were found in the treated sites throughout the study. Di-rhamnolipid (VCRC B426) formulation was also found to reduce the immature density in the treated sites up to 14 days in experiment 1 and up to 15 days in experiment 2. Interpretation & conclusion: For VCRC B471, the optimum application dosage determined was 51 mL/m2 and for VCRC B426, 27mL/m2 . The formulations are to be applied fortnightly for effective control of Anopheles. The application dosage determined in the present study can be used for large scale field evaluation to assess their suitability for use in public health programmes for the control of Anopheles mosquitoes vectoring malaria

Comparison of nutrient-rich and limited media in the production of biosurfactant by Achromobacter xylosoxidans BP(1)5

Aims@#This study aims to produce Achromobacter biosurfactant in nutrient-rich and nutrient-limited media.@*Methodology and results@#This study conducted fermentation on nutrient-rich and nutrient-limited media using a minimal salt medium (MSM). Dextrose and sodium citrate were used as sole carbon supplemented with 0.5% yeast extract for nutrient-rich media, while nutrient-limited media used molasses and rice straw hydrolysate (RSH) at variations of concentrations of 100 ppm and 200 ppm. The research was performed over 120 h and evaluated from growth response, surface tension and emulsification activity. The study revealed that the best surface tension value was when 2% (w/v) sodium citrate was used as C-source and 0.5% (w/v) yeast extract as N-source, after 72 h upon incubation at 30 °C/120 rpm having 45.45 ± 2.19 mN/m with emulsification activity 24.54 ± 3.42%. Whereas the best result of the nutrient-limited medium was obtained by RSH at a concentration of 200 ppm having 48.86 ± 5.36 mN/m.@*Conclusion, significance and impact of study@#The experiment showed that nutrient-limited medium from rice straw hydrolysate could compete with the nutrient-rich medium. The use of rice straw will contribute to the reduction of biosurfactant production costs and valorisation of agricultural waste.

Characterization and Production of Rhamnolipid Biosurfactant in Recombinant Escherichia coli Using Autoinduction Medium and Palm Oil Mill Effluent

Abstract Rhamnolipid is a potent biodegradable surfactant, which frequently used in pharmaceutical and environmental industries, such as enhanced oil recovery and bioremediation. This study aims to engineer Escherichia coli for the heterologous host production of rhamnolipid, to characterize the rhamnolipid product, and to optimize the production using autoinduction medium and POME (palm oil mill effluent). The construction of genes involved in rhamnolipid biosynthesis was designed in two plasmids, pPM RHLAB (mono-rhamnolipid production plasmid) and pPM RHLABC (di-rhamnolipid production plasmid). The characterization of rhamnolipid congeners and activity using high-resolution mass spectrometry (HRMS) and critical micelle concentration (CMC). In order to estimate rhamnolipid yield, an oil spreading test was performed. HRMS and CMC result show E. coli pPM RHLAB mainly produced mono-rhamnolipid (Rha-C14:2) with 900 mg/L and 35.4 mN/m of CMC and surface tension value, whereas E. coli pPM RHLABC mainly produced di-rhamnolipid (Rha-Rha-C10) with 300 mg/L and 34.3 mN/m of CMC and surface tension value, respectively. The optimum condition to produce rhamnolipid was at 20 h cultivation time, 37 oC, and pH 7. In this condition, the maximum rhamnolipid yield of 1245.68 mg/L using autoinduction medium and 318.42 mg/L using 20% (v/v) of POME. In conclusion, the characteristics of the rhamnolipid by recombinant E. coli is very promising to be used in industries as the most economical way of producing rhamnolipid.

Extraction and antimicrobial activity of rhamnolipid biosurfactant produced by Pseudomonas aeruginosa UKMP14T

Aims@#Rhamnolipids are seeking utmost attention as a new class of biosurfactants having promising potential in diverse fields as they offer a wide range of advantages over chemically synthesised surfactants. However, the high extraction costs make large scale production face difficulty. In present study, hydrocarbon degrading bacteria Pseudomonas aeruginosa UKMP14T was exploited for its biosurfactant producing ability including a comparative study between different extraction procedures for its recovery. In addition to this, the recovered biosurfactant was explored for its potential application as an antimicrobial agent. @*Methodology and results@#The production of rhamnolipid biosurfactant was confirmed through various detection methods which are drop-collapse test, oil spreading assay, emulsification index, cetyltrimethylammonium bromide (CTAB) assay and hemolytic assay. The test strain P. aeruginosa UKMP14T showed positive results for all the detection assays. Following this, shake flask cultivation was carried out for several time intervals (1, 3, 5, 7 and 9 days) to discover the optimum time for rhamnolipid biosurfactant production. The results were evaluated by quantifying the rhamnolipid yield using Anthrone method and maximum yield was obtained on day 7. Then, three commonly employed rhamnolipid biosurfactant extraction methods (acid precipitation, solvent extraction and zinc sulphate precipitation) were incorporated for the extraction of rhamnolipid biosurfactant. Among these methods, organic solvent extraction (using methanol, chloroform and acetone in 1:1:1 ratio) gave the highest yield (7.37 ± 0.81 g/L) of biosurfactant, followed by zinc sulphate precipitation (5.83 ± 0.02 g/L), whereas acid precipitation gave the lowest yield (2.8 ± 0.12 g/L) and required longer time (30 days). Finally, the antimicrobial activity of several concentrations of rhamnolipid was tested using modified microdilution method and highest antibacterial activity (in the form of percent reduction in growth) of 95.05% and 91.89% was recorded for Escherichia coli ATCC 10536 and Staphylococcus aureus ATCC 11632, respectively, at 100 µg/mL concentration of rhamnolipid biosurfactant.@*Conclusion, significance and impact of study@#The ability of P. aeruginosa UKMP14T in producing rhamnolipid biosurfactant was confirmed. Despite the higher yield obtained by organic solvent extraction method, the recovery technique (involving the separation of solvent system) caused some loss in product. In addition, the transfer and storage of rhamnolipid was challenging using solvent extraction in comparison to acid precipitation and zinc sulphate precipitation. On the other hand, recovery using acid precipitation suffered from lowest yield of rhamnolipid. Therefore, zinc sulphate precipitation is prioritised over the other two methods. Furthermore, the antimicrobial potential of rhamnolipid biosurfactant was tested successfully for as low as 10 µg/mL concentration against E. coli ATCC 10536 and S. aureus ATCC 11632. Therefore, the recovery cost of a high value product like rhamnolipid can be reduced by incorporating the results of this study in the downstream processing and promote rhamnolipid biosurfactant as a potential antimicrobial agent.

Biosurfactant and bioemulsifier as promising molecules produced by Mucor hiemalis isolated from Caatinga soil

BACKGROUND: The present study describes the production of biosurfactant (BS) and emulsifier (BE) by the filamentous fungus Mucor hiemalis UCP 0039, as well as the characterization and stability of the both biomolecules for environmental or industrial applications. RESULTS: Biosurfactants and bioemulsifiers are amphiphilic compounds and are produced as extracellular molecules. The results showed that bioproduct obtained by shaker condition reduced the water surface tension of 72 to 32 mN/m and reached an emulsification index of 96%, while the static cultivation resulted in a biomolecule with a surface tension of 40 mN/m and an emulsification index of 96%, suggesting the production of a biosurfactant and bioemulsifier, respectively. The compounds showed glycolipid nature but the biosurfactant presented cationic charge, while the bioemulsifier, anionic charge. Thus, the results confirmed that M. hiemalis produced two distinct biomolecules under different parameters and in the same culture medium. CONCLUSIONS: It is the first time that biosurfactant and emulsifier production has been described in the same medium and under different physical conditions by Mucor hiemalis. Both biomolecules showed thermal stability, as well as have significant effect on the viscosity of hydrophobic compounds, indicating the excellent potential for environmental safety or industrial applications to improve the efficiency of sustainable and economic technologies.

Biorremediação de solos contaminados com arsênio por meio de lavagem de solo usando biossurfactantes / Bioremediation of arsenic-contaminated soil through soil washing using biosurfactants

RESUMO A mineração de ouro é uma das principais fontes de contaminação de arsênio (As) no Quadrilátero Ferrífero em Minas Gerais. O As é um elemento tóxico capaz de causar sérios danos à saúde humana. Com o objetivo de investigar formas mais ecológicas e igualmente eficientes para a remoção de As em solos, empregou-se a técnica de lavagem de solos (soil washing) com uso de extratos com biossurfactantes (BS) visando promover a mobilização do As contido em solo de mina de ouro abandonada. Para a produção dos extratos, foram selecionadas duas culturas mistas de bactérias produtoras de BS (MPCB e MPBR) tolerantes a alta concentração de arsênio (8 mg.L-1). Os tensoativos produzidos foram estáveis em ampla faixa de pH, 3-11; temperatura, 28-50ºC e salinidade, 1-5% NaCl (p.v-1). Nos testes de lavagem de solo, em pH 11, as remoções de As obtidas com os extratos MPCB (14,01 e 13,72%) e MPBR (12,04 e 12,31%) foram superiores àquelas obtidas com soluções a 1% (p.v-1) dos surfactantes comerciais SDS (0,87 e 0,71%); saponina (0,57 e 0,55%) e lecitina de soja (2,05 e 2,63%). Os resultados dos testes em coluna foram igualmente influenciados pelo pH e pela proporção sólido:líquido. As maiores remoções de As: 25,43% (MPCB) 22,43% (MPBR) foram obtidas em pH 11, na proporção 1:40 (g.mL-1), após 10 ciclos de extração. Os extratos MPCB e MPBR removeram o As solúvel em água, os íons ligados ao carbonato e aqueles adsorvidos. Ambos os extratos tiveram comportamento semelhante ao ácido etilenodiaminotetracético (EDTA) e ao extrato de cultura microbiana comercial (GorduraKlin®).

ABSTRACT Gold mining is one of the main sources of Arsenic (As) contamination in Minas Gerais, particularly at the Iron quadrangle. Arsenic is a toxic element that causes damage to human health. The soil washing technique using biosurfactants (BS) containing extracts was tested to promote the mobilization of the As contained in some soil samples collected in an abandoned gold mine, to investigate an environmentally friendly but equally efficient alternative to remove As from soils. Two mixed cultures of biosurfactant-producing bacteria (MPCB and MPBR), both adapted to the cultivation at high As concentrations (8 mg.L-1), were selected for extracts production. Surfactants were stable over a wide pH range, 3-11; temperature, 28-50ºC and salinity, 1-5% NaCl (p.v-1), and promoted the mobilization of As. The pH and solid:liquid ratio influenced the removal efficiency of the tested extracts. The highest removals were obtained at pH 11.0 and a solid:liquid ratio of 1:40 (g.mL-1). Under pH 11.0, the arsenic removal results obtained with MPCB (14.01 and 13.72%) and MPBR extracts (12.04 and 12.31%) were superior to those obtained by using 1% (w/v) solutions of the commercial surfactants SDS (0.87 and 0.71%), saponin (0.57 and 0.55%) and, soy lecithin (2.05 and 2.63%). The pH and the solid: liquid ratio influenced the results of the column tests. The highest As removals: 25.43% (MPCB) and 22.43% (MPBR) were obtained after 10 extraction cycles, at pH 11 and 1:40 ratio (g.mL-1). The extracts MPCB and MPBR removed the water-soluble arsenic as well as the carbonate associated As and the adsorbed ions. Both extracts behaved similarly to EDTA and the commercial microbial extract (GorduraKlin®).

Biosurfactant produced by Candida utilis UFPEDA1009 with potential application in cookie formulation

BACKGROUND: Biosurfactants are biomolecules that have the potential to be applied in food formulations due to their low toxicity and ability to improve sensory parameters. Considering the ability of yeasts to produce biosurfactants with food-friendly properties, the aim of the present study was to apply a biosurfactant produced by Candida utilis in the formulation of cookies. RESULTS: The biosurfactant was obtained with a yield of 24.22 ± 0.23 g/L. The characterization analysis revealed that the structure of a metabolized fatty acid with high oleic acid content (68.63 ± 0.61%), and the thermogravimetric analysis demonstrated good stability at temperatures lower than 200°C, potential for food applications. The biosurfactant also exhibited satisfactory antioxidant activity at concentrations evaluated, without cytotoxic potential for cell strains, L929 and RAW 264.7, according to the (3-(4,5-dimethylthiazol-2- yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The incorporation of the surfactant into the dough of a standard cookie formulation to replace animal fat was carried out, achieving a softer, spongier product without significantly altering the physical and physicochemical properties or energy value. CONCLUSION: The thermal stability and antioxidant activity of the biosurfactant produced by C. utilis were verified, besides the positive contribution in the texture analysis of the cookies. Therefore, this biomolecule presents itself as a potential ingredient in flour-based sweet food formulations.

Isolation and characterization of hydrocarbon-degrading bacteria from gas station leaking-contaminated groundwater in the Southern Amazon, Brazil / Isolamento e caracterização de bactérias degradadoras de hidrocarbonetos de água subterrânea contaminada por vazamentos em posto de combustível no sul da Amazônia, Brasil

Abstract Twenty-three hydrocarbon-degrading bacteria strains were isolated from gas station leaking-contaminated groundwater located in the Southern Amazon, Brazil. Based on hydrocarbon (diesel, hexadecane, benzene, toluene and xylene) degradation ability, two strains were selected for further study. The amplification and sequencing of the 16S rRNA gene showed that these two strains belonged to the genus Bacillus (Bacillus sp. L26 and Bacillus sp. L30). GC-MS analysis showed that strain L30 was the most effective in degrading n-alkane (C10-C27) from diesel after 7 days of cultivation in mineral medium. Both strains produced biosurfactants and showed emulsification activity, specially the strain L30. Alkane hydroxylase gene (group III), which is important for alkane biodegradation, was present in strains. As a result, this study indicated that these bacteria could have promising applications in hydrocarbon bioremediation.

Resumo Vinte e três linhagens bacterianas degradadoras de hidrocarbonetos foram isoladas de água subterrânea contaminada por vazamento em posto de combustível no sul da Amazônia, Brasil. Com base na habilidade de degradar hidrocarbonetos (diesel, hexadecano, benzeno, tolueno e xileno), duas linhagens foram selecionadas para estudos posteriores. A amplificação e sequenciamento do gene 16S rRNA demonstrou que essas linhagens pertencem ao gênero Bacillus (Bacillus sp. L26 and Bacillus sp. L30). Análises de GC-MS mostraram que a linhagem L30 foi mais eficiente em degradar n-alcanos (C10-C27) presentes no diesel, após 7 dias de cultivo em meio mineral. Ambas as linhagens produziram biossurfactantes e apresentaram atividade emulsificante, especialmente a linhagem L30. O gene alcano hidroxilase (grupo III), o qual é importante para degradação de alcanos, foram detectados nas linhagens. Como resultado, este estudo indicou que essas linhagens bacterianas podem ser promissoras se aplicadas em processos de biorremediação.

Antibacterial Activity of Sophorolipids from Candida bombicola Against Human Pathogens

Abstract Sophorolipids are glycolipids that have natural antimicrobial properties and present great potential in the pharmaceutical field. The present study aimed to produce sophorolipids from Candida bombicola using a chicken fat-based medium and evaluate the antimicrobial activity against Gram-negative (Proteus mirabilis, Escherichia coli, Salmonella enterica subsp. enterica) and Gram-positive bacteria (Enterococcus faecium, Staphylococcus aureus and Streptococcus mutans). The production of sophorolipids reached 27.86 g L-1. Based on the structural characterization, 73.55% of the sophorolipids present a mixture of acidic monoacetylated C18:2 and lactonic diacetylated C16:0, and 26.45% were present in the diacetylated C18:1 lactonic form. Bacteria submitted to sophorolipid exposure showed a reduction in viability at doses of 500 μg mL-1 and 2,000 μg mL-1 against Gram-positive and Gram-negative bacteria, respectively. These results suggest that sophorolipids produced in chicken fat medium may be used as antimicrobial agents to prevent or eliminate contamination by different pathogens.

Effect of Cultural Conditions on Biosurfactant Production by Candida sp. Isolated from the Sap of Elaeis guineensis

Aims: This study is aimed at determining the effect of cultural condition on biosurfactant production by Candida sp. isolates from saps of Elaeis guineensis. Methodology: Chemical analysis of the sap was carried out. Yeast isolates from the sap were screened for biosurfactant production based on emulsification index (E24), emulsification assay, haemolytic assay, oil displacement test, CTAB and tilted glass slide ability. The best biosurfactant-producing yeast isolate was identified based on its phenotypic, microscopic, and biochemical characteristics. The emulsification capacity of the produced biosurfactant on selected oils was studied. Optimum cultural and nutritional requirements (temperature, pH, inoculum concentration, nitrogen sources and carbon sources) for biosurfactant production by the isolate were determined. Results: The characteristics of the sap from Elaeis guineensis were reducing sugar (0.51 ± 0.03 mg/ml), alcohol (14.04 ± 0.15%), specific gravity (0.827±0.024), and pH (5.68±0.03). The crude biosurfactant produced displaced a thin film of crude oil on petri dish by 55 mm, and revealed high emulsification index (E24) of 52.5% using Olive oil as substrate compared to E24 of 60.6% by sodium dodecyl sulphate (SDS). Based on colonial, microscopic, and biochemical characteristics, the isolate SA2 was identified as Candida sp. The crude biosurfactant showed varying capacity in emulsifying the different oils that were examined. Optimization data revealed maximum biosurfactant production after 7 days of incubation, inoculum concentration of 10%, at temperature of 20ºC, pH of 2 with cassava peel as substrate. Conclusion: The study has demonstrated the capacity of Candida sp. from the sap of Elaeis guineensis to produce biosurfactant utilizing cassava peel as substrate. The use of cassava peel, which represents a low-cost substrate, is important in reducing the cost of biosurfactant production. Moreover, using yeasts from Elaeis guineensis make the production process ecologically friendly.

Characterization of glycolipid biosurfactant from Pseudomonas aeruginosa PA 1 and its efficacy against Phytophtora infestans

Aim: The aim of the present study was to identify biosurfactant produced by Pseudomonas aeruginosa and to determine the effectiveness of biosurfactants (rhamnolipid) against P. infestans causing late blight of potato. Methodology: Pseudomonas aeruginosa strains were isolated from soil samples and the potential strain PA 1, selected because of its antagonistic ability, was used to optimize anti-metabolite production and its characterization using HPLC-Mass spectrometry. Rhamnolipid based formulation was developed and its efficacy was tested against late blight disease. Results: The results revealed that four rhamnolipids congeners were identified, among them three were mono-rhamno-di-lipidic congeners and one was di-rhamno-di-lipdic congeners, abundantly present in the crude biosurfactant obtained from P. aeruginosa PA1. The mass spectra of mono-rhamno-di-lipidic Rha-C12-C14 peak value (m/z 584), Rha-C12.1-C10.CH3 peak value (m/z 545), Rha-C12-C-12-CH3 peak value (m/z 575) and di-rhamno-di-lipidic. Rha-Rha-C10-C10 peak value (m/z 651) were also detected. Rhamnolipid-based formulation was developed and evaluated at different concentration ranging from 0.012 and 0.3% in detached leaf test. Significant reduction in lesion area was recorded at 0.2% concentration (lesion area 0.06 cm2 as against 9.8 cm2 on 5th day of inoculation). Interpretation: Microbial produced rhamnolipid based formulation at 0.2% concentration was found effective against late blight of potato in detached leaf test. Further, it could be used in field study as green chemical which would help in replacing application of chemicals in agriculture.

Low toxicity and high surface activity of sophorolipids from Starmerella bombicola in aquatic species: A preliminary study

Aim: This study was carried out to evaluate aquatic toxicity and surface activity of sophorolipids extracted from yeast Starmerella bombicola. Methodology: The half maximal effective concentration (EC50) values of sophorolipids were determined in three aquatic species, and the surface activity of sophorolipids in aqueous solutions was examined at EC50 concentration. Results: The EC50 of sophorolipid surfactant >473 mg l-1 was found in alga Pseudokirchneriella subcapitata, a median lethal concentration (LC50) of 64.8 mg l-1 in fish Oryzias latipes, whereas a high EC50 of 48.2 mg l-1 was noted in crustacean Daphnia magna, respectively. Sophorolipids effectively reduced the aquatic surface tension to 39 mN m-1 in O. latipes and to 41 mN m-1 in D. magna. Interpretation: These results show that sophorolipids from S. bombicola are a promising biosurfactant with an ideal balance of low aquatic toxicity and high surface activity. Key words: Aquatic toxicity, Biosurfactant, Sophorolipid, Surfactant

Conversion of renewable substrates for biosurfactant production by Rhizopus arrhizus UCP 1607 and enhancing the removal of diesel oil from marine soil

BACKGROUND: The use of agro-industrial wastes to produce high value-added biomolecules such as biosurfactants is a promising approach for lowering the total costs of production. This study aimed to produce biosurfactants using Rhizopus arrhizus UCP 1607, with crude glycerol (CG) and corn steep liquor (CSL) as substrates. In addition, the biomolecule was characterized, and its efficiency in removing petroderivatives from marine soil was investigated. RESULTS: A 22 factorial design was applied, and the best condition for producing the biosurfactant was determined in assay 4 (3% CG and 5% CSL). The biosurfactant reduced the surface tension of water from 72 to 28.8 mN/m and produced a yield of 1.74 g/L. The preliminary biochemical characterization showed that the biosurfactant consisted of proteins (38.0%), carbohydrates (35.4%), and lipids (5.5%). The compounds presented an anionic character, nontoxicity, and great stability for all conditions tested. The biomolecule displayed great ability in dispersing hydrophobic substrates in water, thereby resulting in 53.4 cm2 ODA. The best efficiency of the biosurfactant in removing the pollutant diesel oil from marine soil was 79.4%. CONCLUSIONS: This study demonstrated the ability of R. arrhizus UCP1607 to produce a low-cost biosurfactant characterized as a glycoprotein and its potential use in the bioremediation of the hydrophobic diesel oil pollutant in marine soil

Biosurfactant from endophytic Bacillus cereus: Optimization, characterization and cytotoxicity study

Aims@#The increased importance of biosurfactant in the recent past is mainly due to their applications in various industries ranging from petroleum to pharmaceuticals. Their biodegradability and environmental compatibility with low toxicity makes it even more interesting. Microbial production of biosurfactant is found to be a viable option as they are diverse, eco-friendly, facilitate large scale production, able to perform under extreme conditions etc. One class of microbes that is endophytes are known to show great potential in producing different varieties of medically and industrially significant biological compounds. The present study focuses on the screening and production of biosurfactant from endophytic bacteria. @*Methodology and results@#Of all the isolates tested, one endophyte identified as Bacillus cereus HM998898 was found to produce maximum biosurfactant. Statistical method Plackett burman was used to optimize the media for the maximum production and the ideal composition was found to be KNO3 (1 g/L), Gingley oil (2 mL), K2HPO4 (2.5 g/L), KH2PO4 (0.75 g/L), MgSO4·5H2O (0.5 g/L), FeSO4.7H2O (0.005 g/L) and NaCl (0.025 g/L). The extracted biosurfactant was characterized and was identified to be glycolipid. This was further tested for biocompatibility against Fibroblast (3T3) cells and was evaluated for their anti tumor activity against Hep2 cells. @*Conclusion, significance and impact of study@#The biosurfactant produced was found to induce toxicity to cancer cells at appreciable levels while they remained non-toxic to normal cells supporting the possible applications of biosurfactant in medical field.

Production and characterization of a biosurfactant produced by Streptomyces sp. DPUA 1559 isolated from lichens of the Amazon region

Surfactants are amphipathic compounds containing both hydrophilic and hydrophobic groups, capable to lower the surface or interfacial tension. Considering the advantages of the use of biosurfactants produced by microorganisms, the aim of this paper was to develop and characterize a biosurfactant produced by Streptomyces sp. DPUA1559 isolated from lichens of the Amazon region. The microorganism was cultured in a mineral medium containing 1% residual frying soybean oil as the carbon source. The kinetics of biosurfactant production was accompanied by reducing the surface tension of the culture medium from 60 to values around 27.14 mN/m, and by the emulsification index, which showed the efficiency of the biosurfactant as an emulsifier of hydrophobic compounds. The yield of the isolated biosurfactant was 1.74 g/L, in addition to the excellent capability of reducing the surface tension (25.34 mN/m), as observed from the central composite rotational design when the biosurfactant was produced at pH 8.5 at 28°C. The critical micelle concentration of the biosurfactant was determined as 0.01 g/mL. The biosurfactant showed thermal and pH stability regarding the surface tension reduction, and tolerance under high salt concentrations. The isolated biosurfactant showed no toxicity to the micro-crustacean Artemia salina, and to the seeds of lettuce (Lactuca sativa L.) and cabbage (Brassica oleracea L.). The biochemistry characterization of the biosurfactant showed a single protein band, an acid character and a molecular weight around 14.3 kDa, suggesting its glycoproteic nature. The results are promising for the industrial application of this new biosurfactant.

Characterization of dioxygenases and biosurfactants produced by crude oil degrading soil bacteria

ABSTRACT Role of microbes in bioremediation of oil spills has become inevitable owing to their eco friendly nature. This study focused on the isolation and characterization of bacterial strains with superior oil degrading potential from crude-oil contaminated soil. Three such bacterial strains were selected and subsequently identified by 16S rRNA gene sequence analysis as Corynebacterium aurimucosum, Acinetobacter baumannii and Microbacterium hydrocarbonoxydans respectively. The specific activity of catechol 1,2 dioxygenase (C12O) and catechol 2,3 dioxygenase (C23O) was determined in these three strains wherein the activity of C12O was more than that of C23O. Among the three strains, Microbacterium hydrocarbonoxydans exhibited superior crude oil degrading ability as evidenced by its superior growth rate in crude oil enriched medium and enhanced activity of dioxygenases. Also degradation of total petroleum hydrocarbon (TPH) in crude oil was higher with Microbacterium hydrocarbonoxydans. The three strains also produced biosurfactants of glycolipid nature as indicated d by biochemical, FTIR and GCMS analysis. These findings emphasize that such bacterial strains with superior oil degrading capacity may find their potential application in bioremediation of oil spills and conservation of marine and soil ecosystem.

Kinetic study and modeling of biosurfactant production using Bacillus sp

Background: Surfactants are one of the most important raw materials used in various industrial fields as emulsifiers, corrosion inhibitors, foaming agents, detergent products, and so on. However, commercial surfactant production is costly, and its demand is steadily increasing. This study aimed to evaluate the performance of typical strains of Bacillus sp. to produce biosurfactants through fermentation. It also included the investigation of the effect of initial glucose concentration and the carbon to nitrogen ratio. Results: The biosurfactant yield was in the range of 1­2.46 g/L at initial glucose concentrations of 10­70 g/L. The optimum fermentation condition was achieved at a carbon to nitrogen ratio of 12.4, with a decrease in surface tension of up to 27 mN/m. Conclusions: For further development and industrial applications, the modified Gompertz equation is proposed to predict the cell mass and biosurfactant production as a goodness of fit was obtained with this model. The modified Gompertz equation was also extended to enable the excellent prediction of the surface tension.

Produção biotecnológica de biossurfactante por Lactococcus lactis CECT-4434 a partir de resíduos agroindustriais e avaliação de suas propriedades / Biotechnological production of biosurfactant by Lactococcus lactis CECT- 4434 from agroindustrial residues and evaluation of its properties

A bactéria Lactococcus lactis subsp. lactis CECT-4434 foi empregada para investigar o efeito da composição do meio de cultivo na produção biotecnológica de biossurfactante e, adicionalmente bacteriocina. Utilizou-se resíduos agroindustriais, tais como soro de leite e vinhaça de uva, para formular meios de cultivos mais econômicos e naturais, suplementados sacarose e extrato de levedura. Um planejamento fatorial fracionado 24, com adição de três ensaios nos pontos centrais foi empregado para avaliar a influência destas variáveis. A produção de biossurfactante foi influenciada positivamente pela concentração soro de leite, onde 15 % deste demonstrou melhor resultado reduzindo a tensão superficial em cerca de 18,1 mN/m, alcançando produção máxima de biossurfactante equivalente em surfactina de 11,02 mg/L. Em relação à síntese de bacteriocina, a fonte de carbono adicional (sacarose) interferiu de forma antagonista, ou seja, quanto menor a concentração de sacarose, maior a síntese de bacteriocina (com aumento da zona de inibição em 14,2% contra Staphylococcus aureus CECT-239). Observou-se que o ensaio conduzido em biorreator, sob microaeração com 5% de oxigênio dissolvido, promoveu maior produção de biossurfactante (11,6 mg/L) quando comparados aos estudos conduzidos com maior concentração de oxigênio entre 30 a 100%, com produção em média de 2,3 mg/mL. Destaca-se que nenhum estudo da influência do oxigênio dissolvido, principalmente em microaerofilia, para a produção de biossurfactante por bactérias láticas já havia sido realizado. Ademais, o biossurfactante produzido se mostrou altamente estável frente a valores extremos de pH e temperatura, além de demonstrar notável propriedade antimicrobiana e antiadesiva, inibindo Listeria monocytogenes NADC 2045 e Salmonella entérica CECT-724 em mais de 90%.

Lactococcus lactis subsp. lactis CECT-4434 was used to investigate the effect of the composition of the culture media on the biotechnological production of biosurfactant and bacteriocin additionally. Agroindustrial residues, such as whey and grape vinasse, were used to formulate more economical and natural culture media, supplemented with sucrose and yeast extract. A fractional factorial design 24, with addition of three runs at the central points was used to evaluate the influence of these variables. The biosurfactant production was positively influenced by the concentration of whey, where 15% showed a better result reducing the surface tension by 18.1 mN/m, reaching a maximum production of biosurfactant equivalent in surfactin of 11.02 mg/L. In relation to bacteriocin synthesis, the sucrose interfered in an antagonistic way, that is, the lower the sucrose concentration, the greater the bacteriocin synthesis (with an increase in the zone of inhibition in 14.2% against Staphylococcus aureus CECT-239). It was observed that the bioreactor conducted under microaeration with 5% dissolved oxygen promoted a higher biosurfactant production (11.6 mg/L) when compared to studies conducted with a higher concentration of oxygen between 30 and 100%, with production on average 2.3 mg/mL. It is noteworthy that no study of the influence of dissolved oxygen, mainly in microaerophilic, for the biosurfactant production by lactic acid bacteria had already been carried out. In addition, the biosurfactant produced proved to be highly stable against extreme values of pH and temperature, and demonstrated remarkable antimicrobial and antiadhesive properties, inhibiting Listeria monocytogenes NADC 2045 and Salmonella entérica CECT-724 in more than 90%.