Production Of Bioemulsifiers Research Articles

Exploring the potential of novel thermophilic bacterial strain for the production of bioemulsifiers using the hydrolysate of sugarcane bagasse

Exploring the potential of novel thermophilic bacterial strain for the production of bioemulsifiers using the hydrolysate of sugarcane bagasse

Sustainable production of bioemulsifiers, a critical overview from microorganisms to promising applications

Microbial bioemulsifiers are molecules of amphiphilic nature and high molecular weight that are efficient in emulsifying two immiscible phases such as water and oil. These molecules are less effective in reducing surface tension and are synthesized by bacteria, yeast and filamentous fungi. Unlike synthetic emulsifiers, microbial bioemulsifiers have unique advantages such as biocompatibility, non-toxicity, biodegradability, efficiency at low concentrations and high selectivity under different conditions of pH, temperature and salinity. The adoption of microbial bioemulsifiers as alternatives to their synthetic counterparts has been growing in ongoing research. This article analyzes the production of microbial-based emulsifiers, the raw materials and fermentation processes used, as well as the scale-up and commercial applications of some of these biomolecules. The current trend of incorporating natural compounds into industrial formulations indicates that the search for new bioemulsifiers will continue to increase, with emphasis on performance improvement and economically viable processes.

Molecular characterization of Paenibacillus antarcticus IPAC21, a bioemulsifier producer isolated from Antarctic soil.

Paenibacillus antarcticus IPAC21, an endospore-forming and bioemulsifier-producing strain, was isolated from King George Island, Antarctica. As psychrotolerant/psychrophilic bacteria can be considered promising sources for novel products such as bioactive compounds and other industrially relevant substances/compounds, the IPAC21 genome was sequenced using Illumina Hi-seq, and a search for genes related to the production of bioemulsifiers and other metabolic pathways was performed. The IPAC21 strain has a genome of 5,505,124 bp and a G + C content of 40.5%. Genes related to the biosynthesis of exopolysaccharides, such as the gene that encodes the extracellular enzyme levansucrase responsible for the synthesis of levan, the 2,3-butanediol pathway, PTS sugar transporters, cold-shock proteins, and chaperones were found in its genome. IPAC21 cell-free supernatants obtained after cell growth in trypticase soy broth at different temperatures were evaluated for bioemulsifier production by the emulsification index (EI) using hexadecane, kerosene and diesel. EI values higher than 50% were obtained using the three oil derivatives when IPAC21 was grown at 28°C. The bioemulsifier produced by P. antarcticus IPAC21 was stable at different NaCl concentrations, low temperatures and pH values, suggesting its potential use in lower and moderate temperature processes in the petroleum industry.

Production and Characterization of a Bioemulsifier Derived from Microorganisms with Potential Application in the Food Industry.

There is a growing interest in the development and use of natural emulsifiers, which provide biodegradability as well as non-toxicity along with giving better performance compared to existing emulsifying agents used in the food industry. A large variety of sources of starting material, i.e., the microorganisms, are available to be used, hence giving a diverse range of applications. The focus of this review paper is on the production of bioemulsifiers, which are said to be “green surfactants”, from fungi, bacteria and yeasts; furthermore, an overview pertaining to the knowledge gained over the years in terms of characterization techniques is reported. The methods used for the characterization and isolation such as TLC, GC-MS, HPLC, NMR have also been studied. The end-application products such as cookies, muffins, and doughs along with the methods used for the incorporation of bioemulsifiers, microorganisms from which they are derived, properties imparted to the product with the use of a particular bioemulsifier and comparison with the existing food grade emulsifiers has been discussed in detail. The future prospects indicate that newer bioemulsifiers with anti-microbial, anti-oxidant and stabilization properties will prove to have a larger impact, and emphasis will be on improving the performance at an economically viable methodology.

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 of a novel thermophilic bacterium capable of producing high-yield bioemulsifier and its kinetic modelling aspects along with proposed metabolic pathway

Bioemulsifiers form stable emulsions and lower surface tension between two phases with potent anti-microbial activities. Some applications of bioemulsifier are performed at high temperatures and hence production of bioemulsifiers that are stable at high temperature is required. This study aimed at the production of bioemulsifier by an unexplored bacterial strain isolated from a local hot spring. The parameters tested for bioemulsifier production (emulsification ability, surface tension measurement and product formation) showed that 24h is the optimal time for the production of bioemulsifier by strain S3 with yield of 1.4g/l. The logistic growth curve of bacterial strain was analysed and kinetic constants for substrate utilisation and product formation were determined by Luedeking-Piret kinetic models. The bacterial strain S3 was Gram-positive and was classified as a strain of Brevibacillus borstelensis. The specific growth rate of the organism was 0.0096h-1 with the kinetic rate constants as 11.246 (γ) and 10.626 (δ) for Luedeking-Piret substrate and 3.8423 (α) and - 1.9075 (β) for Luedeking-Piret product. Knowledge of these values will help in estimating the substrate utilisation or bioemulsifier formed at any time point. These studies will also help in understanding internal metabolic fluxes hence rigorous analysis of metabolic pathway of bioemulsan is also performed in this study. Graphical abstract.

Comparative data on effects of alkaline pretreatments and enzymatic hydrolysis on bioemulsifier production from sugarcane straw by Cutaneotrichosporon mucoides

Bioemulsifiers are surface active compounds which could be potentially used in food processing, cosmetic sector and oil recovery. Sugarcane straw (SS), was used as the raw substrate for the production of bio-emulsifiers (BE) by Cutaneotrichosporon mucoides. Three different delignification strategies using dilute sodium hydroxide, sodium sulfite and ammonium hydroxide followed by enzymatic hydrolysis (Cellic CTec 2, 7.5% total solids, 15 FPU/g, 72 h) were studied. Enzyme hydrolysis of ammonium hydroxide pretreated SS showed a maximum of 62.19 ± 0.74 g/l total reducing sugars with 88.35% hydrolytic efficiency (HE) followed by sodium hydroxide (60.06 ± 0.33 g/l; 85.40% HE) and sodium sulfite pretreated SS (57.22 ± 0.52 g/l; 84.71% HE), respectively. The ultrastructure of SS (native and delignified) by fourier transform-infrared and near infrared spectroscopy, revealed notable structural differences. The fermentation of hydrolysates by C. mucoides into bioemulsifiers showing emulsification index (EI) of 54.33%, 48.66% and 32.66% from sodium sulfite, sodium hydroxide, and ammonium hydroxide pretreated SS, respectively.

Production and characterization of bioemulsifiers from Acinetobacter strains isolated from lipid-rich wastewater.

In this study, two indigenous bacterial strains (Ab9-ES and Ab33-ES) isolated from lipid-rich wastewater showed potential to produce bioemulsifier in the presence of 2% (v/v) olive oil as a carbon source. These bacterial strains were identified as Acinetobacter sp. Ab9-ES and Acinetobacter sp. Ab33-ES by polymerase chain reaction and analysis of 16S rRNA gene sequences. Bioemulsifier production by these strains was found to be growth-linked. Maximum emulsifying activities (83.8% and 80.8%) were recorded from strains Ab9-ES and Ab33-ES, respectively. Bioemulsifier yields of 4.52g/L and 4.31g/L were obtained from strains Ab9-ES (XB9) and Ab33-ES (YB33), respectively. Fourier-transform infrared spectroscopic analysis revealed the glycoprotein nature of the bioemulsifiers. The bioemulsifiers formed stable emulsions only in the presence of edible oils. Maximum emulsifying activities of 79.6% (XB9) and 67.9% (YB33) were recorded in the presence of sunflower oil. The bioemulsifiers were found to be stable at a broad range of temperature (4-121°C), moderate pH (5.0-10.0) and salinity (1-6%). In addition, bioemulsifier XB9 showed maximum emulsifying activities (77.3%, 74.5%, and 74.9%) at optimum temperature (50°C), pH (7.0), and NaCl concentration (3%), respectively. On the contrary, YB33 demonstrated highest activities (73.6%, 72%, and 61.2%) at optimum conditions of 70°C, pH 7.0, and NaCl concentration of 5%, respectively. Findings from this study suggest the potential biotechnological applications of the bioemulsifiers, especially in the remediation of oil-polluted sites.

Evaluation of a microbial consortium for crude oil spill bioremediation and its potential uses in enhanced oil recovery

Abstract Microbial enhanced oil recovery (MEOR) process utilizes reservoir microorganisms (in situ) or specially selected bacteria to use their metabolic products (ex-situ) for extra oil recovery from depleted oil reservoirs. Biodegradation of crude oil and simultaneous production of bioemulsifiers by a mixed culture consisting of Ochrobactrum pseudintermedium C1 and Bacillus cereus K1 have been investigated in this study along with the potential ex-situ application of bioemulsifier in MEOR process. The culture parameters namely pH, temperature, aeration rate and agitation rate have been optimized in a lab scale bioreactor using Taguchi's L9 orthogonal array approach to increase the rate of biodegradation of crude oil. At optimized culture condition (pH 8.0, temperature 35 °C, aeration rate 1.5Nl/min and agitation rate 150 rpm), the bacterial consortium degrade up to 70.54% of total petroleum hydrocarbons (TPH) after 72 h incubation using crude oil (4% v/v) as the sole source of carbon and produced two different exopolysaccharides (EPS), which showed very good emulsification activity (EI) towards crude oil up to 80.5% and also reduced the interfacial tension of crude-oil water system from 46 mN/m to 14.5 mN/m. FT-IR, NMR and chemical analyses revealed that EPS formed by C1 contained 61.5% (w/w) carbohydrates, 29.2% protein and 9.3% lipid whereas, the EPS formed by K1 comprises of 43.6% (w/w) carbohydrates, 51.7% lipids and 6.3% protein. The crude bioemulsifier mixture was found to be pseudoplastic, Non-Newtonian in nature and was stable at a wide range of pH (2−10), temperature (40–121 °C) and salinity (0–15% w/v), hence signifies its applicability in MEOR process. The results of the sand pack column flooding tests at simulated reservoir conditions demonstrated that the additional oil recovery efficiency due to the ex-situ injection of the cell-free bioemulsifier solution was 40.93% and 46.85% at 40 °C and 70 °C temperatures respectively. The mixed culture presented a great potential application in bioremediation of oil polluted sites owing to its degradation ability to crude oil and also in enhanced oil recovery process using the crude bioemulsifier having combined surface and emulsification activity.

Production, characterization, and applications of bioemulsifiers (BE) and biosurfactants (BS) produced by Acinetobacter spp.: A review.

Bioemulsifiers (BE) and biosurfactants (BS) are considered as multifunctional biomolecules of 21st century because of their functional abilities and eco-friendly properties. They are produced by various microorganisms under versatile and extreme environmental conditions. They have tremendous applications in various industries such as petroleum, food, medicine, pharmaceutical, chemical, paper & pulp, textile, and cosmetics. Currently, they are also considered as "green molecules" because of their wide applications in bioremediation of soil. Their importance has been increasing day by day in the global market as they are the natural resources with high-aggregate value. Although, there are numerous reports on BE and BS production by different bacteria, Acinetobacter spp. acquired special attention among all. This is because it is the earliest member known for the production of bioemulsifier. Emulsan and Alasan are the best examples of the commercially used BE produced by Acinetobacter spp. These BE are mainly used in microbial enhanced oil recovery and biodegradation of toxic compounds. This review is focused on BE and BS produced by Acinetobacter spp., their characterization and applications in different fields. This is the first review on genus Acinetobacter which defines independently about different types of BE and BS produced by it. It will also address the need of exploration of these molecules from various sources and their applications for the benefit of mankind and sustainable environment.

Production of a microbial emulsifier with biotechnological potential for environmental applications.

The present work reports the production of bioemulsifiers (BEs) by an environmental bacterium closely related to Bacillus spp., using agro-industrial wastes and by-products as low-cost carbon sources. Maximum emulsifying activity was detected using crude glycerol (CG) (E24 = 59%), producing 2.8 g/L of BE at 24 h of incubation. The microbial product presented significantly higher biodegradability in comparison to three commercial emulsifying agents (sodium dodecyl sulfate, SDS; Tween 80; and Arabic gum, AG). Interestingly, BE proved to be innocuous for Caco-2 cells and wheat seedlings, used as toxicological indicators. The BE and AG showed (in most of the cases) higher stability to changes in temperature (37 °C-100 °C), pH (3-10), and salt concentration (5% and 10%, w/v) in comparison to the SDS and Tween 80. Finally, the microbial product displayed a large affinity to a wide range of hydrophobic substrates showing emulsifying activities similar to or even better than SDS, Tween 80 and AG. The results presented in this study demonstrate the potential of a bioproduct obtained from CG to be used for environmental purposes.

Lab scale fermentation studies with marine bacteria to maximize the production of bioemulsifiers for cosmetic and food industries

Lipases are essentially described as sn-1 and sn-3 regio-selective. Actually few methods are available to measure this lipase regio-selectivity, moreover they require chiral chromatography analysis or specific derivations which are discontinuous and time consuming. In this study we describe a new, convenient, sensitive and continuous spectrophotometric method to screen lipases regio-selectivity using synthetic triglycerides (TG) containing α-eleostearic acid (9Z, 11E, 13E-octadecatrienoic acid) either at the sn-1 position [1-α-eleostearoyl-2,3-octadecyl-sn-glycerol (sn-EOO)] or at the sn-3 position [1,2-octadecyl-3-α-eleostearoyl-sn-glycerol (sn-OOE)] and coated onto the wells of microtiter plates. A non-hydrolysable ether bond, with a non UV-absorbing alkyl chain, was introduced at the other sn positions to prevent acyl chain migration during TG synthesis or lipolysis. The synthesis of TG containing α-eleostearic acid was performed from S-glycidol in six steps to obtain sn-EOO and in five steps to sn-OOE. The α-eleostearic acid conjugated triene constitutes an intrinsic chromophore and, consequently, confers the strong UV absorption properties of this free fatty acid as well as of the TG harboring it. The lipase activity on coated sn-EOO or sn-OOE was measured by the increase in the absorbance at 272 nm due to the transition of α-eleostearic acid from the adsorbed to the soluble state. Human and porcine pancreatic lipases, guinea pig pancreatic lipase related protein 2, Thermomyces lanuginosus lipase, Candida antarctica lipase A and Candida antarctica lipase B were all used to validate the assay. This continuous high-throughput screening method could determine directly without any processes after lipolysis the regio-selectivity of various lipases.

Stable bioemulsifiers are produced by Acinetobacter bouvetii UAM25 growing in different carbon sources.

Acinetobacter species are identified as producing surface-active and emulsifying molecules known as bioemulsifiers. Production, characterization and stability of bioemulsifiers produced by Acinetobacter bouvetii UAM25 were studied. A. bouvetii UAM25 grew in three different carbon and energy sources: ethanol, a glycerol-hexadecane mixture and waste cooking oil in an airlift bioreactor, showing that bioemulsifier production was growth associated. The three purified bioemulsifiers were lipo-heteropolysaccharides of high molecular weight (4866 ± 533 and 462 ± 101kDa). The best carbon source and energy for bioemulsifier production was wasted cooking oil, with a highest emulsifying capacity (76.2 ± 3.5EUmg-1) as compared with ethanol (46.6 ± 7.1EUmg-1) and the glycerol-hexadecane mixture (49.5 ± 4.2EUmg-1). The three bioemulsifiers in our study displayed similar macromolecular structures, regardless of the nature (hydrophobic or hydrophilic) of the carbon and energy source. Bioemulsifiers did not decrease surface tension, but the emulsifying capacity of all of them was retained under extreme variation in salinity (0-50gNaClL-1), pH (3-10) and temperature (25-121°C), indicative of remarkable stability. These findings contribute to understanding of the relationship between: production, physical properties, chemical composition and stability of bioemulsifiers for their potential applications in biotechnology, such as bioremediation of hydrocarbon-contaminated soil and water.

Isolation of hydrocarbon-degrading and biosurfactant-producing bacteria and assessment their plant growth-promoting traits

Forty-two hydrocarbon-degrading bacterial strains were isolated from the soil heavily contaminated with petroleum hydrocarbons. Forty-one strains were identified based on their whole-cell fatty acid profiles using the MIDI-MIS method. Thirty-three of them belong to species Rhodococcus erythropolis, while the others to the genera Rahnella (4), Serratia (3) and Proteus (1). Isolates were screened for their ability to produce biosurfactants/bioemulsifiers. For all of them the activity of several mechanisms characteristic for plant growth-promoting bacteria was also determined. In order to investigate surface active and emulsifying abilities of isolates following methods: oil-spreading, blood agar, methylene blue agar and determination of emulsification index, were used. Among studied bacteria 12 strains (CD 112, CD 126, CD 131, CD 132, CD 135, CD 147, CD 154, CD 155, CD 158, CD 161, CD 166 and CD 167) have been chosen as promising candidates for the production of biosurfactants and/or bioemulsifiers. Among them 2 strains (R. erythropolis CD 126 and Rahnella aquatilis CD 132) had the highest potential to be used in the bioaugmentation of PH-contaminated soil. Moreover, 15 of tested strains (CD 105, CD 106, CD 108, CD 111, CD 116, CD 120, CD 124, CD 125, CD 130, CD 132, CD 134, CD 154, CD 156, CD 161 and CD 170) showed the activity of four mechanisms (ACC deaminase activity, IAA and siderophore production, phosphate solubilization) considered to be characteristic for plant growth-promoting bacteria. Two of them (R. erythropolis CD 106 and R. erythropolis CD 111) showed the highest activity of above-mentioned mechanisms and thus are considered as promising agents in microbe assisted phytoremediation.

Properties of Polyhydroxyalkanoate Granules and Bioemulsifiers from Pseudomonas sp. and Burkholderia sp. Isolates Growing on Glucose.

A Burkholderia and Pseudomonas species designated as AB4 and AS1, respectively, were isolated from soil containing decomposing straw or sugar cane bagasse collected from Brazil. This study sought to evaluate the capacities of culture media, cell-free medium, and crude lysate preparations (containing PHB inclusion bodies) from bacterial cell cultures to stabilize emulsions with several hydrophobic compounds. Four conditions showed good production of bioemulsifiers (E24 ≥ 50%), headed by substantially cell-free media from bacterial cell cultures in which bacterial isolates from Burkholderia sp. strain AB4 and Pseudomonas sp. strain AS1 were grown. Our results revealed that the both isolates (AB4 and AS1 strains) exhibited high emulsification indices (indicating usefulness in bioremediation) and good stabilities.

Retracted Article: Bigger data open innovation: potential applications of value-added products from milk and sustainable valorization of by-products from the dairy industry

Milk is a natural suspension of essential nutrients and can be exploited not only as a beverage consumed directly or transformed into a range of traditional dairy foods, but also as a source of ingredients for food product innovation. In addition, by-products from the dairy industry, which used to constitute an economical and environmental problem, are still rich in proteins, lactose and minerals. Therefore, from a green point of view, they can be valorized and are being exploited for multiple food and non-food applications. This paper aims to give an overview of the different applications of milk ingredients and by-products from the dairy industry. Additionally, enzymatic modification and production of bioactive peptides and bioemulsifiers broaden their functionality and applicability. Finally, a critical review of the most promising aspects as well as some features that need further development is presented. The unmet needs, the cross fertilization in between protein domains, the carbon footprint requirements, the environmental necessities, the health and wellness new demands, etc. are dominant factors in the search for an innovation approach outlining the potential that these “apparent” constraints oblige science and technology to account for. The adjacent technology analysis opportunity that such a review generates is, in essence, an open innovation step.

Polyhydroxyalkanoate Synthesis Affects Biosurfactant Production and Cell Attachment to Hydrocarbons in Pseudomonas sp. KA-08

Stressful conditions prevailing in hydrocarbon-contaminated sites influence the diversity, distribution, and activities of microorganisms. Oil bioremediation agents should develop special characteristics to cope with these environments like surfactant production and cellular affinity to hydrocarbons. Additionally, polyhydroxyalkanoate (PHA) accumulation was proven to improve tolerance to stressful conditions. Pseudomonas sp. KA-08 was isolated from a chronic oil-contaminated environment, it is highly tolerant to xylene, and it is able to accumulate PHA and to produce surfactant compounds that lower the water surface tension (ST) as well as bioemulsifiers. In this work, we studied the effect of the capability to accumulate PHAs on biosurfactant production and microbial attachment to hydrocarbons (MATH). Our results showed that PHA synthesis capability has a favorable effect in the production of compounds which affect the ST but not on the production of bioemulsifiers. On the other hand, PHA accumulation affects cellular affinity to xylene. MATH analysis showed that a PHA-negative mutant increased its affinity to xylene compared with the wild-type strain. This result was also observed in Pseudomonas putida GPp104 (a PHA(-) mutant), suggesting that this effect could be generalized to other Pseudomonas strains.

Production of bioemulsifiers by Amycolatopsis tucumanensis DSM 45259 and their potential application in remediation technologies for soils contaminated with hexavalent chromium

In recent years, increasing interest has been shown in the use of bioemulsifiers as washing agents that can enhance desorption of soil-bound metals. However, high production costs derived from the use of expensive substrates for formulation of the fermentation media represent the main challenge for full, large-scale implementation of bioemulsifiers. This work reports on a first study of bioemulsifier production by the actinobacterium Amycolatopsis tucumanensis DSM 45259 using different carbon and nitrogen sources. Preliminary results on the potential use of these compounds as washing agents for soils contaminated with Cu(II) and Cr(VI) are also presented. The best specific production was detected using glycerol and urea as carbon and nitrogen substrates, respectively. However, with all of the substrates used during the batch assay, the bioemulsifiers showed high levels of stability at extreme conditions of pH, temperature, and salt concentration. Under the current assay conditions, the bioemulsifiers were not effective in removing Cu(II) from soil. However, they were able to mediate Cr(VI) recovery, with the removal percentage doubled compared to that seen when using deionized water. These findings appear promising for the development of remediation technologies for hexavalent chromium compounds based upon direct use of these microbial emulsifiers.

Production of bioemulsifiers by Yarrowia lipolytica in sea water using diesel oil as the carbon source

The objective of this work was to investigate, on a flask scale, the production of bioemulsifiers by Yarrowia lipolytica in the presence of sea water, supplemented with nitrogen and phosphate sources, using diesel oil as substrate. A full 24 factorial design was conducted to investigate the effects and interactions of the nutrient concentrations (diesel oil, urea, ammonium sulfate and monobasic potassium phosphate) on the response variables: emulsification activity and surface tension of the cell-free cultures. High emulsification activities (> 5,4 UEA) were determined after 168 h in all the experiments. The interactions among diesel oil, urea and monobasic potassium phosphate favored the emulsification with statistical significance. A correlation between the increase of emulsification activity and the reduction of surface tension was not identified.

Production and Characterization of Bioemulsifiers by Thermotolerant Bacteria for Enhanced Oil Recovery Potential

Three bacterial strains were isolated from oil contaminated soil samples in Yumen oilfield of China and selected due to their capacity of growing under extreme conditions and production of bioemulsifier. The isolates were identified as Brevibacillus sp. XS1, Geobacillus sp. XS2 and Geobacillus sp. XS3 according to 16S rDNA sequence and physiological methods, respectively. The isolates XS1, XS2 and XS3 were thermotolerant with the optimum growth temperature of 50 °C, 60 °C and 55 °C, respectively. All the three isolates were able to produce bioemulsifiers which had little effect on surface tension reduction. The bioemusifiers produced by the three isolates were purified with a production yield of 2.98g/L, 4.24g/L and 3.82 g/L, respectively. The bioemulsifiers were identified as anionic heteropolysaccharides by FTIR analysis and their average molecular weight and polydispersity were investigated by GPC method. The bioemulsifiers produced by Geobacillus sp. XS2 and XS3 exhibited a perfect stability over various temperature (up to 100 °C), pH (range from 2.0 to 12.0) and salinity (up to 30.0% of NaCl concentration) and were considered to be ideal candidates in MEOR process.