Isolation and characterization of biosurfactant producing bacteria from Mile 2 and Ologe Lagoons, Nigeria

Aims@#This study was aimed to isolate and characterize biosurfactant producing bacteria from Mile 2 and Ologe Lagoon which are sinks for domestic and industrial waste waters and potential source of value added bioresources such as biosurfactants, hydrocarbon degraders and organisms with potential for biotechnological applications.@*Methodology and results@#Physicochemical parameters of the two lagoon waters were analyzed using standard procedures. Bacteria were isolated using enrichment techniques on 1% Escravos light crude oil, palm oil and groundnut oil on mineral salt medium (MSM). Biosurfactant production by the isolates was assayed by hemolytic activity, oil spread test, blue agar test and emulsification activity. Isolates were identified using their colony morphologies and biochemical characteristics, while the antibiotic susceptibility of the isolates was determined using multidisc. The physicochemistry of the lagoon water showed high nitrate content of 15.7 mg/L and 19.6 mg/L for Mile 2 and Ologe Lagoon, respectively. Total hydrocarbon content (THC) of both lagoon waters was low, with values 0.53 mg/L for Mile 2 Lagoon and 0.44 mg/L for Ologe Lagoon. The predominant genera of bacteria identified include Micrococcus, Bacillus, Pseudomonas, Acinetobacter, Stomatococcus and Moraxella. A total of 23 bacterial isolates were tested for hemolytic activity, of which 13 showed β-hemolysis which is presumptive for biosurfactant production, 5 showed α-hemolysis and the remaining 5 exhibited γ-hemolysis. Majority of the isolates were positive for oil spread assay and blue agar test (19) indicating production of anionic biosurfactant. The isolates showed good emulsification activity; AGG3 (67.7%), AGG1 (62.3%), AGG2 (60%), AGG4 (60%), MTP2 (56%), AGC4 (54%) and the least emulsification value of 23.3% for strain AGP1. Most of the isolates were susceptible to ciprofloxacin, perfloxacin and showed resistance to septrin and erythromycin.@*Conclusion, significance and impact of study@#This study showed that Mile 2 and Ologe Lagoon are a potential source of biosurfactant producers with diverse emulsification properties and prospective industrial applications. This would have implication for economic empowerment, as well as sustainable and environmentally friendly clean-up technology in both locally and globally.

Butter whey and corn steep liquor as sole raw materials to obtain a bioemulsifier from Yarrowia lipolytica for food oil-in-water emulsions / Soro de manteiga e milhocina como únicas matérias primas para obter bioemulsificante de Yarrowia lipolytica para emulsões óleo-em-água alimentícias

ABSTRACT: A synthetic medium containing glucose, glycerol, yeast extract (YE), and ammonium sulfate (AS) was compared to several low-cost media in their ability to produce high emulsification index (EI). The goal was to reduce the production costs of an emulsifier with application in food oil-in-water emulsions. To this end, agro-industrial by-products were screened for bioemulsifier production from Yarrowia lipolytica. The statistical analysis showed that the EIs of media containing residual frying oil from palm oil (RFO_palm) or soybean oil (RFO_soy), residual liquid from butter production (butter whey, BWhey) or cheese production (cheese whey, CWhey), supplemented with YE and AS were similar to the EI of the synthetic medium. The replacement of YE by corn steep liquor (CSL) also resulted in similar EI, except for RFO_soy. BWhey was tested with CSL without AS and similar EI (66.8%) was detected in comparison to that of the same medium with AS (66.3%). The cell-free broth obtained after Y. lipolytica growth in BWhey+CSL was successfully used to obtain vegetable oil-in-water emulsions indicating its potential application in food products.

RESUMO: O meio sintético contendo glicose, glicerol, extrato de levedura (YE) e sulfato de amônio (AS) foi comparado a meios de baixo custo para produzir alto índice de emulsificação (EI). Para este fim, os subprodutos agroindustriais foram rastreados quanto à produção de bioemulsificante por Yarrowia lipolytica. A análise estatística mostrou que EI de meios contendo óleo de fritura residual de óleo de palma (RFO_palm) ou óleo de soja (RFO_soy), líquido residual da produção de manteiga (soro de manteiga, BWhey) ou da produção de queijo (soro de queijo, CWhey), suplementado com YE e AS foram semelhantes ao EI do meio sintético. A substituição do YE por milhocina (CSL) também resultou em EI semelhante, exceto no RFO_soy. O BWhey foi testado com CSL sem AS e EI semelhante (66,8%) foi detectado em comparação com o mesmo meio com AS (66,3%). O meio isento de células obtido após o crescimento de Y. lipolytica no meio BWhey + CSL foi utilizado com sucesso para obter emulsões de óleo vegetal em água, indicando sua potencial aplicação em produtos alimentícios.

Optimisation Of Biosurfactant Production By Serratia Rubidaea Kap Isolated From Oil Contaminated Soil Sample

Surfactants are surface active molecules which play an important role in variousindustrial processes and product formation due to their interfacial properties, biodegradabilityand eco-friendly nature. Many of the chemical surfactants in use today are produced fromnon-renewable petrochemical feedstocks, hence bio surfactants produced by microorganismsusing renewable and cheap substrates are considered as viable alternatives to petroleumbased surfactants. In the current study, the optimization of various parameters for maximumproduction of bio surfactant, by Serratia rubidaea KAP isolated from oil contaminated soilsample, was carried out. Optimum bio surfactant production was obtained in a mediumcontaining; (g %) KH2PO4 (0.3), Na2HPO4 (0.6), yeast extract (0.5), NaCl (0.5), (NH4)2SO4 (0.3),MgSO4.7H2O (2mg), FeSO4.7H2O (0.002), and CaCl2.2H2O (0.006), rice bran oil (5% v/v) with C:N and C: P ratio as 8:1 and 7:1 respectively. The media was adjusted to pH 7.2, inoculated with5% v/v test culture prepared at 0.8O.D600nm, and incubated at 30°C for 96h under agitationspeed of 160rpm. The resulting bio surfactant yield of 1.0g% was achieved, with 0.88g% weightof dry biomass, 56.78% decrease in surface tension of the medium and emulsification index of87.5%. Thus the isolate showed good potential as a bio surfactant-producing bacterium thatcan be used for large scale production using rice bran oil as a cost-effective substrate, as analternative to chemical surfactants.

Physicochemical screening of Candida lusitaniae P1 during synthesis of biosurfactant from coconut shell

Aims: Fermentation and recovery are the major operating cost in biosurfactant production. Thus, the aim of this research was to synthesize biosurfactant from agricultural residues using suitable fermentation and recovery techniques in order to reduce the cost of production. Methodology and results: Biosurfactant-producing yeast strains isolated from refined oil-contaminated soil samples using yeast extract-diesel agar (YEDA) were subjected to physicochemical screening such as drop collapse test, microplate analysis, oil spreading technique, emulsification index and thermostability. Based on the preliminary screening result, Candida lusitaniae P1, C. parapsilosis P51, C. parapsilosis D3 and C. lusitaniae E1 were selected for biosurfactant production using agricultural residue such as rice bran, wheat bran and coconut shell as substrate and crude supernatant was analyzed by gas chromatography mass spectrometry. Candida lusitaniae P1 strain produced 98.96 g/L of biosurfactant from coconut shell but when subjected to mutagenesis the yield decreased to 52.24 g/L. Conclusion, significance, and impact of study: The physicochemical properties of biosurfactant produced using various carbon sources showed that coconut-shell is the best residue thus, variation in composition and concentration of biosurfactant obtained implies that the quality and quantity of biosurfactant produced depends on the carbon source and the genetic composition of the yeast isolate.

Biodegradation of Phenol and Emulsification Properties of Native Microorganisms from Coal Carbonization Plant.

Phenols (aromatic organic compounds) are commonly occurring organic pollutants in the environment. Some of the toxic effects of phenol in humans are corrosive to eyes, skin and respiratory track, in addition also causes lethal effects in animal and plant community when discharged even in low concentrations. The problem associated with phenol in the environment, is in need to removal such hazardous compound. Considering the importance, the present investigation attempts for degradation of phenol by native bacterial strains isolated from coal carbonization plant wastewater. About 36 bacterial isolates were found to exhibit phenol degrading activity. Among them, higher phenol degrading strains were identified tentatively as CA1 – Rhodococcus sp., CA2 – Bacillus sp., CA3 – Pseudomonas sp. and CA4 –Citrobacter sp. Phenol degrading ability of Rhodococcus sp. was found to be 60% with maximum cell dry weight. Surface active compounds were observed to be 1.31G for Rhodococcus sp, 0.85 G for Bacillus sp, 1.26 G for Pseudomonas sp. and 1.02 G for Citrobacter sp. In case of mixed cultures, 1.62 G maximum rate among the tested was observed. The Emulsification Index (E24) for CA1, CA2, CA3, CA4 and mixed culture was observed as 78%, 65%, 59%, 22% and 80% respectively. The plasmid profile was additionally studied for the isolates. Significantly, the study reveals the potentials of native strain, Rhodococcus sp. (CA1), in degradation of phenols.

Evaluation of growth in diesel fuel and surfactants production ability by bacteria isolated from fuels in Costa Rica / Evaluación de crecimiento en combustible diesel y capacidad de producción de surfactantes en bacterias aisladas de combustibles en Costa Rica

A total of 149 bacterial strains previously isolated from fuels stored in Costa Rica were selected in terms of their ability to grow aerobically in diesel and produce bioemulsifier active compounds. The diesel growth was evaluated by a redox-indicator based test, and surfactant production was estimated indirectly by both the emulsification index determination (E24) and hemolytic activity. Twenty-six strains (16.8%) were considered as capable of growing in diesel, while surfactant production was detected in 22 (14.8%), estimated according to E24. Seven strains showed high production of biosurfactants (E24 ≥ 50%), headed by Pseudomonas aeruginosa 148D-O, P. aeruginosa 87R-B and Bacillus pumilus 133S-B. No significant correlation was observed between hemolytic patterns and growth outcomes in diesel or E24. Surfactant producing strains should be studied further to assess its potential applications.

Un total de 149 cepas bacterianas previamente aisladas de combustibles almacenados en Costa Rica fueron seleccionadas en términos de sus habilidades para crecer aeróbicamente en diesel y producir compuestos con actividad bioemulsificante. El crecimiento en diesel fue evaluado por medio de un test basado en un indicador redox, y la producción de surfactantes fue estimada indirectamente con las determinaciones del índice de emulsificación (E24) y la actividad hemolítica. Veintiseis cepas (16,8%) fueron consideradas como capaces de crecer en diesel, mientras que la producción de surfactantes fue detectada en 22 (14,8%), estimado de acuerdo con el E24. Siete cepas mostraron alta producción de biosurfactantes (E24 ≥ 50%), encabezadas por Pseudomonas aeruginosa 148D-O, P. aeruginosa 87R-B y Bacillus pumilus 133S-B. No se observó correlación significativa entre los patrones de hemólisis y los resultados de crecimiento en diesel o E24. Las cepas productoras de surfactantes deben ser estudiadas más a fondo para evaluar sus potenciales aplicaciones.

Effect of nutrients and fermentation conditions on the production of biosurfactants using rhizobacteria isolated from fique plants / Efecto de los nutrientes y las condiciones de fermentación en la producción de biosurfactantes con rizobacterias aisladas de fique / Efeito dos nutrientes e as condições de fermentação na produção de biossurfactantes com rizobacté rias isoladas da piteira

Objective. To isolate biosurfactant-producing microorganisms from the rhizosphere of fique and to select the best genus to evaluate the effect of nutritional and fermentation conditions on the production of rhamnolipids. Materials and methods. Rhizospheric soil was sampled in three areas of Cauca. The best genus was selected for the experimental designs (Plackett Burman and 2² factorial) and to find the production conditions for the growth kinetics at an Erlenmeyer flask scale. Results. Isolates from the rhizosphere of fique plants were from groups (or genera) of Bacillus, Pseudomonas and Actinomycetes, being Pseudomonas the more responsive in preliminary testing for emulsification. From the results of the experimental designs and the kinetics of production, we found that rhamnose synthesis associated with rhamnolipids (3.2 g/l) and emulsification (68% EC24) was significantly favored (p <0.0001) by cultivating an inoculum of 10% v/v of Pseudomonas fluorescens in a medium composed of: soybean oil 2% (v/v), K2HPO4 0.2% (w/v), yeast extract 0.4 g/l, NH4NO3 3.7 g/l, 1 ml trace elements (CoCl3 20 mg/l, H3BO(3)30 mg/l, ZnSO(4)10 mg/l, Cu2SO(4)1 mg/l, Na2MoO(4)3 mg/l, FeSO (4)10 mg/l MnSO(4)2,6 mg/l) and pH 7.2. Conclusion. Of all the microbial genera isolated from the rhizosphere of fique, Pseudomonas fluorescens had the greatest potential in the production of biosurfactants of the rhamnolipids family.

Objetivo. Aislar microorganismos de la rizosfera de fique capaces de producir biosurfactantes y seleccionar el mejor gé nero para evaluar el efecto de las condiciones nutricionales y de fermentación en la producción de ramnolípidos. Materiales y mé todos. Se realizaron muestreos de suelos rizosfé ricos en tres zonas del Cauca. El mejor gé nero fue seleccionado para realizar los diseños experimentales (Plackett Burman y factorial 2²) y establecer las condiciones de producción para las ciné ticas de crecimiento a escala de Erlemeyer. Resultados. Se aislaron bacterias del gé nero Bacillus, Pseudomonas y del grupo Actinomycetes, siendo Pseudomonas el grupo con mayor respuesta en las pruebas preliminares de emulsificación. A partir de los resultados obtenidos en los diseños experimentales y ciné ticas de producción, se estableció que la síntesis de ramnosa asociada con ramnolípidos (3,2 g/l) y la emulsificación (68% EC24) se favorecieron significativamente (p<0.0001) al cultivar un inoculo de 5% v/v de Pseudomonas fluorescens en un medio compuesto por: aceite de soya 2% (v/v), K2HPO4 0,2% (p/v), extracto de levadura 0,4 g/l, NH4NO(3)3,7 g/l, 1 ml de elementos traza (CoCl(3)20 mg/l, H3BO(3)30 mg/l, ZnSO(4)10 mg/l, Cu2SO4 1 mg/l, Na2MoO(4)3 mg/l, FeSO(4)10 mg/l MnSO(4)2,6 mg/l) y pH 7.2. Conclusión. Se aislaron 3 gé neros microbianos a partir de rizosfera de fique, siendo Pseudomonas fluorescens la bacteria con mayor potencial en la producción de biosurfactantes de la familia de los ramnolípidos.

Objetivo. Isolar microorganismos da rizosfera da piteira capazes de produzir biossurfactantes, selecionar o melhor gênero para avaliar o efeito das condições nutricionais e de fermentação na produção de rhamnolipídeos. Materiais e mé todos. Foram realizadas amostragem de solos rizosfé ricos em três á reas do Cauca. O melhor gênero foi selecionado para realizar desenhos experimentais (Plackett Burman e fatorial 2²) e definir as condições de produção para as ciné ticas em escala de erlenmeyer. Resultados. Foram isoladas bacté rias do gênero Bacillus, Pseudomonas e grupo de Actinomycetes. As Pseudomonas foram o grupo com maior resposta nos testes preliminares de emulsificação. A partir dos resultados obtidos nos desenhos experimentais e ciné ticas de produção foi estabelecido que a síntese de ramnose associados com rhamnolipídeos (3,2 g/l) e a emulsificação (68% EC24) foram favorecidos significativamente (p<0,0001) ao cultivar um inoculo de 5% v/ v de Pseudomonas fluorescens em um meio composto por: óleo de soja 2% (v/v), K2HPO(4)0,2% (p/v), extrato de levedura 0,4 g/l, NH4NO3 3,7 g/l, 1 ml de oligoelementos (CoCl(3)20 mg/l, H3BO(3)30 mg/l, ZnSO(4)10 mg/l, Cu2SO(4)1 mg/l, Na2MoO(4)3 mg/l, FeSO(4)10 mg/l MnSO4 2,6 mg/l) e pH 7,2. Conclusão. Três gêneros microbianos foram isolados da rizosfera da piteira, sendo Pseudomonas fluorescens a bacté ria com maior potencial na produção de biossurfactantes da família dos rhamnolipídeos.