Optimization of Growth Parameters for Enhancing the Production of Biosurfactants from Pediococcus pentosaceus S-2

Numerous Lactic acid bacteria (LAB) have been found to be capable of synthesizing surface-active compounds i.e biosurfactants. These are amphiphilic compounds produced by microorganisms on their cell surface or secreted extracellularly that have a tendency to reduce surface and interfacial tension. In the present study, different process parameters including nitrogen and carbon source, pH, temperature, aeration and agitation were optimized to maximize the production of biosurfactants from Pediococcus pentosaceus S-2. Xylose (1.5%) and yeast extract (1.5%) act as better carbon and nitrogen sources respectively for the production of biosurfactants. Maximum biosurfactant yield was observed at pH 6, a temperature of 35o C, an agitation rate of 200 rpm and with inoculum size of 3%. The high yield of biosurfactants produced from Pediococcus pentosaceus S-2 by utilizing media supplemented with whey under optimized conditions.

Benchmarking of different microbes for their biosurfactants antifungal action against plant pathogens

The biotic stress caused by phytopathogens (bacteria, fungus, yeast and insect pests) is a primary factor in yield loss of plants. Biocontrol agents and their active compounds are used to manage such plant pathogens. Here, in our study, we screened four bacterial isolates identified as Bacillus cereus, B. anthracis, B. velezensis and Serratia marcescens after morphological, biochemical and molecular characterization (16s rDNA sequencing) for production of biosurfactant by foam forming activity, oil spreading tests and emulsification activity. Highest foam stability (75 min) and maximum emulsification activity E24% (75%) was observed by B. velezensis strain. Among all the four isolates, Bacillus velezensis strain produced maximum biosurfactant (0.349±0.004 g/50 mL). Biosurfactant of all the four bacterial isolates were checked for fungal inhibiton on PDA plate(s). Bacillus velezensis showed comparatively the highest percent inhibition 58.82, 88.15, 78.45,72.68, 83.96, 75.47, 68.07 and 88.44% against Colletotrichum falcatum, Fusarium oxysporum f sp. ciceri, Helminthosporium maydis, F. oxysporum f. sp. lycopersici, Aspergillus niger, Mucor sp., Helminthosporium oryzae and Rhizoctonia solani, respectively. Bacillus velezensis biosurfactant among all the four bacterial isolates was found to be most effective against the tested phytopathogens

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.

Production of green surfactants: market prospects

Science has greatly contributed to the advancement of technology and to the innovation of production processes and their applications. Cleaning products have become indispensable in today's world, as personal and environmental hygiene is important to all societies worldwide. Such products are used in the home, in most work environments and in the industrial sectors. Most of the detergents on the market are synthesised from petrochemical products. However, the interest in reducing the use of products harmful to human health and the environment has led to the search for detergents formulated with natural, biodegradable surfactant components of biological (plant or microbiological) origin or chemically synthesised from natural raw materials usually referred to as green surfactants. This review addresses the different types, properties, and uses of surfactants, with a focus on green surfactants, and describes the current scenario as well as the projections for the future market economy related to the production of the different types of green surfactants marketed in the world.

Biosurfactant Production by Fusarium oxysporum LM 5634 Using Peels from the fruit of Bactris gasipaes (Kunth) as Substrate

Aim: The aim of this work was to find new fungal sources of biosurfactants via bioprospecting and to produce biosurfactants from fungal isolates using inexpensive culture media. Methodology: Bioprocess tests were conducted with five fungal species isolated from soil from the Amazon region in order to investigate which of them presented the highest productivity in a kinetics experiment. The possibility of using the peels from the fruit of Astrocaryum aculeatum Meyer (PFAC), Bactris gasipaes Kunth (PFBG), Theobroma grandiflorum Schumann (PFTG), Musa paradisiaca (PFMP) as substrates was also evaluated. Results: Fusarium oxysporum LM 5634 had the highest productivity, and PFBG was the best substrate for biosurfactant production. Conclusion: Thus, the present study showed the potential of fungi and wastes (fruit peels) from the Amazon region for biosurfactant production.

Ecology and biotechnological potential of bacterial community from three marine sponges of the coast of Rio de Janeiro, Brazil

ABSTRACT Marine sponges has been a large reservoir of microbial diversity, with the presence of many species specific populations as well as producing biologically active compounds, which has attracted great biotechnological interest. In order to verify the influence of the environment in the composition of the bacterial community present in marine sponges and biotechnological potential of bacteria isolated from these organisms, three species of sponges and the waters surrounding them were collected in different beaches of Rio de Janeiro, Brazil. The profile of the bacterial community present in sponges and water was obtained by PCR-DGGE technique and the biotechnological potential of the strains isolated by producing amylase, cellulase, protease and biosurfactants. The results showed that despite the influence of the environment in the composition of the microbial community, studied marine sponges shown to have specific bacterial populations, with some, showing potential in the production of substances of biotechnological applications.

Presencia de genes rhlAB, rhlR y rhlC en Pseudomonas aeruginosa nativas sobreproductoras de ramnolípidos

Se realizó la caracterización molecular de los genes asociados a la producción de ramnolípidos (RL), en 61 cepas bacterianas de la colección del Laboratorio de Microbiología y Biotecnología Microbiana (LAMYBIM) de la Universidad Nacional Mayor de San Marcos, Perú. Las cepas provenian de entornos peruanos contaminados con hidrocarburos y fueron catalogadas como sobreproductoras de RL(n= 21), productoras de RL (n=20) y no productoras de RL (n= 20). Las 61 cepas fueron identificadas bioquímicamente con el sistema API 20 NE. La identificación molecular se realizó empleando el gen del RNAr 16S. Se encontró que Pseudomonas aeruginosa fue el microorganismo de mayor prevalencia en los estratos sobreproductores y productores de ramnolípidos. Además, se encontraron: Burkholderia cepacea, Pseudomonas fluorescens, Aeromonas hydrophila y Chryseobacterium indologenes. Los microorganismos no productores de ramnolípidos, también fueron caracterizados bioquímicamente. Mediante amplificación de PCR y electroforesis en gel de agarosa, estandarizados por la UNAM, se evidenció que las cepas seleccionadas poseen los genes: rhlA, rhlB, rhlR y rhlC. Para el secuenciamiento de la región génica rhLABR, se seleccionaron cuatros especies: Pseudomonas aeruginosa T2X-2, Pseudomonas aeruginosa IIIT1P2, Pseudomonas aeruginosa 6K-11 y Pseudomonas aeruginosa ATCC 9027, siguiendo metodología estandarizada por la UNAM y fueron comparados con Pseudomonas aeruginosa PAO1. Nuestros resultados muestran que los genes estudiados en las cepas seleccionadas son sinónimas de sus homólogos en la cepa patrón Pseudomonas aeruginosa PAO1, por lo que las diferencias genotípicas que expliquen la sobreproducción de ramnolípidos deberían hallarse en otros marcadores moleculares no cubiertos en el presente estudio.

Genes associated to rhamnolipids production were molecularly characterized in 61 bacterial strains from LAMYBIM bacterial collection (Laboratorio de Microbiología y Biotecnología Microbiana, Universidad Nacional Mayor de San Marcos, Perú). Strains were isolated from peruvian environments hydrocarbons polluted and were classified as RL overproducers (n= 21), RL producers (n = 20) and non-producers (n = 20) producers. Molecular identification using the 16S rRNA gene was preceded by the biochemical identification of 61 strains selected with the API 20 NE system. Pseudomonas aeruginosa was the most prevalent strain of the RL overproducers and RL producers. Species such as Burkholderia cepacea, Pseudomonas fluorescens, Aeromonas hydrophila and Chryseobacterium indologenes, were found too. In the same way, non-producers microorganisms were also characterized. The PCR amplification and agarose gel electrophoresis techniques, standarized by the UNAM laboratory, showed that the selected strains had the genes: rhlA, rhlB, rhlR and rhlC. For the sequencing of the rhLABR gene region, four strains were selected: Pseudomonas aeruginosa T2K2, Pseudomonas aeruginosa III T1P2, Pseudomonas aeruginosa 6K-11 and Pseudomonas aeruginosa ATCC 9027, applying the methodology standardized by the UNAM and were compared with Pseudomonas aeruginosa PAO1. Our results show that the genes studied in the selected strains are synonymous with their homologues in Pseudomonas aeruginosa PAO1 standard strain. Therefore, genotypical differences that explain the overproduction of rhamnolipid might be found in other molecular markers not covered in this study.

Optimización y modificación del método para la detección de ramnolípidos

El uso de biosurfactantes en biorremediación facilita y acelera la degradación microbiana de hidrocarburos. El método del agar CTAB/MB creado por Siegmund y Wagner para el screening de cepas productoras de ramnolípidos (RL), ha sido ampliamente utilizado sin sufrir mejoras significativas en más de 20 años. Con el fin de optimizar la técnica como método cuantitativo, se hicieron placas con agar CTAB/MB y se probaron diferentes variables, tales como tiempo de incubación, refrigeración, concentración de CTAB, presencia de azul de metileno, diámetro de los pozos y volumen del inóculo. Adicionalmente, se desarrolló un nuevo método para detectar el RL de los halos mediante precipitación con HCl, lo cual permite la observación de un nuevo patrón de halos más fáciles de observar y medir. Esta investigación reafirma que este método no es del todo apropiado para un análisis cuantitativo fino, debido a la dificultad de correlacionar de forma precisa la concentración de RL y el área de los halos formados. La difusión del RL no parece tener un comportamiento simple y existen numerosos factores que afectan la velocidad de migración del RL.

Use of biosurfactants in bioremediation, facilitates and accelerates microbial degradation of hydrocarbons. CTAB/MB agar method created by Siegmund & Wagner for screening of rhamnolipids (RL) producing strains, has been widely used but has not improved significantly for more than 20 years. To optimize the technique as a quantitative method, CTAB/MB agar plates were made and different variables were tested, like incubation time, cooling, CTAB concentration, methylene blue presence, wells diameter and inocula volume. Furthermore, a new method for RL detection within halos was developed: precipitation of RL with HCl, allows the formation a new halos pattern, easier to observe and to measure. This research reaffirm that this method is not totally suitable for a fine quantitative analysis, because of the difficulty to accurately correlate RL concentration and the area of the halos. RL diffusion does not seem to have a simple behavior and there are a lot of factors that affect RL migration rate.

Properties of a biosurfactant produced by Bacillus pumilus using vinasse and waste frying oil as alternative carbon sources

Biosurfactants are chemical molecules produced by the microorganisms with potential for application in various industrial and environmental sectors. The production parameters and the physicochemical properties of a biosurfactant synthesized by Bacillus pumilus using different concentrations of vinasse and waste frying oil as alternative carbon sources were analyzed. The microorganism was able to grow and produce a biosurfactant using both the residues. The surface tension was reduced up to 45 mN/m and the maximum production of crude biosurfactant was 27.7 and 5.7 g/l for vinasse and waste frying oil, respectively, in concentration of 5%. The critical micelle concentration (CMC) results of 1.5 and 0.2 g/l showed the efficiency of the biosurfactant produced on both the substrates. The results showed that the alternative substrates could be used for the production of an efficient biosurfactant by B. pumilus. These properties have potential for industrial and environmental applications.

Biosurfactants production by yeasts using soybean oil and glycerol as low cost substrate

Biosurfactants are bioactive agents that can be produced by many different microorganisms. Among those, special attention is given to yeasts, since they can produce many types of biosurfactants in large scale, using several kinds of substrates, justifying its use for industrial production of those products. For this production to be economically viable, the use of residual carbon sources is recommended. The present study isolated yeasts from soil contaminated with petroleum oil hydrocarbons and assessed their capacity for producing biosurfactants in low cost substrates. From a microbial consortium enriched, seven yeasts were isolated, all showing potential for producing biosurfactants in soybean oil. The isolate LBPF 3, characterized as Candida antarctica, obtained the highest levels of production - with a final production of 13.86 g/L. The isolate LBPF 9, using glycerol carbon source, obtained the highest reduction in surface tension in the growth medium: approximately 43% of reduction after 24 hours of incubation. The products obtained by the isolates presented surfactant activity, which reduced water surface tension to values that varied from 34 mN/m, obtained from the product of isolates LBPF 3 and 16 LBPF 7 (respectively characterized as Candida antarctica and Candida albicans) to 43 mN/m from the isolate LPPF 9, using glycerol as substrate. The assessed isolates all showed potential for the production of biosurfactants in conventional sources of carbon as well as in agroindustrial residue, especially in glycerol.

Selection of biosurfactan/bioemulsifier-producing bacteria from hydrocarbon-contaminated soil

Petroleum-derived hydrocarbons are among the most persistent soil contaminants, and some hydrocarbon-degrading microorganisms can produce biosurfactants to increase bioavailability and degradation. The aim of this work was to identify biosurfactant-producing bacterial strains isolated from hydrocarbon-contaminated sites, and to evaluate their biosurfactant properties. The drop-collapse method and minimal agar added with a layer of combustoleo were used for screening, and positive strains were grown in liquid medium, and surface tension and emulsification index were determined in cell-free supernantant and cell suspension. A total of 324 bacterial strains were tested, and 17 were positive for the drop-collapse and hydrocarbon-layer agar methods. Most of the strains were Pseudomonas, except for three strains (Acinetobacter, Bacillus, Rhodococcus). Surface tension was similar in cell-free and cell suspension measurements, with values in the range of 58 to 26 (mN/m), and all formed stable emulsions with motor oil (76-93 percent E24). Considering the variety of molecular structures among microbial biosurfactants, they have different chemical properties that can be exploited commercially, for applications as diverse as bioremediation or degradable detergents.

Reducing COD level on oily effluent by utilizing biosurfactant-producing bacteria

Two bacteria isolated from crude oil contaminated soil, Pantoea agglomerans and Planococcus citreus, produced biosurfactants utilizing 1.5 percent of kerosene and olive oil as the sole carbon sources, respectively. The bacteria and the biosurfactants produced were introduced to oily effluent, arising from margarine and soap industry. Emulsification activities were determined by increases in the absorbance of the oil-in-water emulsions at 610 nm, whereas the water-in-oil emulsions were expressed as the height (cm) of the emulsion layers formed. The 72 h incubation experiment resulted in a COD (Chemical Oxygen Demand) reduction of 76 percent with Planococcus citreus strain and 70 percent with Pantoea agglomerans.The COD reduction with bacterial biosurfactants was over 50 percent in 24 h of incubation. The COD reduction showed that these strains and the surfactants produced could be used in bioremediation processes.

Duas bactérias isoladas de solo contaminado com derivados de petróleo, Pantoea agglomerans e Planococcus citreus, produzem biosurfactantes utilizando respectivamente 1.5 por cento de querosene e óleo de oliva como únicas fontes de carbono. As bactérias e os biosurfactantes produzidos foram adicionados a um efluente oleoso obtido de uma indústria nacional de sabão e margarina. As atividades de emulsificação foram determinadas pelo aumento da absorbância das emulsões óleo em água a 610 nm, enquanto que as emulsões do tipo água em óleo foram expressas em centímetros, pela altura do halo de espumas formado. A redução da demanda química de oxigênio (COD) mostra que as linhagens e os biosurfactantes produzidos podem ser utilizados em processos de biorremediação.

OPTIMIZATION OF CONDITIONS FOR FERMENTATION OF BIOSURFACTANT PRODUCED BY PSEUDOMONAS SP. L-11 / 微生物学通报

A strain of pseudomonas sp. L-11 could produce biosurfactants by use of glucose as substrate. The fermentation conditions in shaking flask and 1L fennentor were investigated. The optimal medium and (he fermentation technology were demonstrated. In 1L fermentor, the interracial tension between the diluent of L-11 (10%) and crude oil is 5.3?10~3mN/m, which is fit for microbial enhanced oil recovery. The scale-up technology of L-11 was also studied.