ABSTRACT
Bacteria can synthesize a broad spectrum of multifunctional polysaccharides including extracellular polysaccharides (EPS). Bacterial EPS can be utilized in the food, pharmaceutical, and biomedical areas owing to their physical and rheological properties in addition to generally presenting low toxicity. From an ecological viewpoint, EPS are biodegradable and environment compatible, offering several advantages over synthetic compounds. This study investigated the EPS produced by Klebsiella oxytoca (KO-EPS) by chemically characterizing and evaluating its properties. The monosaccharide components of the KO-EPS were determined by HPLC coupled with a refractive index detector and GC-MS. The KO-EPS was then analyzed by methylation analysis, FT-IR and NMR spectroscopy to give a potential primary structure. KO-EPS demonstrated the ability to stabilize hydrophilic emulsions with various hydrophobic compounds, including hydrocarbons and vegetable and mineral oils. In terms of iron chelation capacity, the KO-EPS could sequester 41.9 % and 34.1 % of the most common iron states, Fe2+ and Fe3+, respectively. Moreover, KO-EPS exhibited an improvement in the viscosity of aqueous dispersion, being proportional to the increase in its concentration and presenting a non-Newtonian pseudoplastic flow behavior. KO-EPS also did not present a cytotoxic effect indicating that the KO-EPS could have potential applications as a natural thickener, bioemulsifier, and bioremediation agent.
Subject(s)
Biodegradation, Environmental , Emulsions , Klebsiella oxytoca , Polysaccharides, Bacterial , Rheology , Klebsiella oxytoca/metabolism , Polysaccharides, Bacterial/chemistry , Polysaccharides, Bacterial/biosynthesis , Emulsifying Agents/chemistry , Emulsifying Agents/metabolism , Biotechnology/methods , Viscosity , Hydrophobic and Hydrophilic InteractionsABSTRACT
Biofilm formation is a major health concern and studies have been pursued to find compounds able to prevent biofilm establishment and remove pre-existing biofilms. While biosurfactants (BS) have been well-known for possessing antibiofilm activities, bioemulsifiers (BE) are still scarcely explored for this purpose. The present study aimed to evaluate the bioemulsifying properties of cell-free supernatants produced by Bacillaceae and Vibrio strains isolated from marine sponges and investigate their antiadhesive and antibiofilm activities against different pathogenic Gram-positive and Gram-negative bacteria. The BE production by the marine strains was confirmed by the emulsion test, drop-collapsing, oil-displacement, cell hydrophobicity and hemolysis assays. Notably, Bacillus cereus 64BHI1101 displayed remarkable emulsifying activity and the ultrastructure analysis of its BE extract (BE64-1) revealed the presence of structures typically observed in macromolecules composed of polysaccharides and proteins. BE64-1 showed notable antiadhesive and antibiofilm activities against Staphylococcus aureus, with a reduction of adherence of up to 100 % and a dispersion of biofilm of 80 %, without affecting its growth. BE64-1 also showed inhibition of Staphylococcus epidermidis and Escherichia coli biofilm formation and adhesion. Thus, this study provides a starting point for exploring the antiadhesive and antibiofilm activities of BE from sponge-associated bacteria, which could serve as a valuable tool for future research to combat S. aureus biofilms.
Subject(s)
Bacterial Adhesion , Biofilms , Emulsifying Agents , Porifera , Staphylococcus aureus , Biofilms/drug effects , Biofilms/growth & development , Porifera/microbiology , Animals , Bacterial Adhesion/drug effects , Staphylococcus aureus/drug effects , Staphylococcus aureus/physiology , Emulsifying Agents/pharmacology , Emulsifying Agents/chemistry , Staphylococcus epidermidis/drug effects , Staphylococcus epidermidis/physiology , Escherichia coli/drug effects , Escherichia coli/physiology , Hydrophobic and Hydrophilic Interactions , Anti-Bacterial Agents/pharmacology , Bacillus cereus/drug effects , Bacillus cereus/physiology , Hemolysis , Surface-Active Agents/pharmacology , Surface-Active Agents/metabolism , Vibrio/drug effects , Vibrio/physiology , Vibrio/metabolism , Microbial Sensitivity Tests , Gram-Negative Bacteria/drug effects , Gram-Negative Bacteria/physiologyABSTRACT
At the best conditions of the bioprocess (30 °C, pH 7.0, 3.0 g/L NaCl) were obtained 0.66 g/L cell concentration, 3.3 g/L of bioemulsifier, which showed high emulsifying activity (53 % ± 2), reducing the surface tension of the water in 47.2 % (38 mN/m). The polymeric structure of the purified bioemulsifier comprised a carbohydrate backbone composed of hexose-based amino sugars with a monomeric mass of 1099 Da, structurally similar to emulsan. A. venetianus bioemulsifier is non-phytotoxic (GI% > 80 %) against Ocimum basilicum and Brassica oleracea and non-cytotoxic (LC50 5794 mg/L) against Artemia salina, being safe local organisms in comparison to other less eco-friendly synthetic emulsifiers. This bioemulsifier effectively dispersed spilled oil in vitro (C22-C33), reducing oil mass by 12 % (w/w) and dispersing oil in a displacement area of 75 cm2 (23.8 % of the spilled area). Thus, the isolated A. venetianus AMO1502 produced a bioemulsifier potentially applicable for environmentally friendly oil spill remediation.
Subject(s)
Acinetobacter , Biodegradation, Environmental , Emulsifying Agents , Acinetobacter/metabolism , Artemia , Animals , Water Pollutants, Chemical , Brassica , Petroleum Pollution , Ocimum basilicumABSTRACT
The demand for emulsion-based products is crucial for economic development and societal well-being, spanning diverse industries such as food, cosmetics, pharmaceuticals, and oil extraction. Formulating these products relies on emulsifiers, a distinct class of surfactants. However, many conventional emulsifiers are derived from petrochemicals or synthetic sources, posing potential environmental and human health risks. In this context, fungal bioemulsifiers emerge as a compelling and sustainable alternative, demonstrating superior performance, enhanced biodegradability, and safety for human consumption. From this perspective, the present work provides the first comprehensive review of fungal bioemulsifiers, categorizing them based on their chemical nature and microbial origin. This includes polysaccharides, proteins, glycoproteins, polymeric glycolipids, and carbohydrate-lipid-protein complexes. Examples of particular interest are scleroglucan, a polysaccharide produced by Sclerotium rolfsii, and mannoproteins present in the cell walls of various yeasts, including Saccharomyces cerevisiae. Furthermore, this study examines the feasibility of incorporating fungal bioemulsifiers in the food and oil industries and their potential role in bioremediation events for oil-polluted marine environments. Finally, this exploration encourages further research on fungal bioemulsifier bioprospecting, with far-reaching implications for advancing sustainable and eco-friendly practices across various industrial sectors.
Subject(s)
Bioprospecting , Cell Wall , Humans , Emulsifying Agents , Food , Glycolipids , Saccharomyces cerevisiaeABSTRACT
Surface-active compounds (SACs) of microbial origin are an active group of biomolecules with potential use in the formulation of emulsions. In this sense, the present study aimed to isolate and select yeasts from fruits that could produce SACs for essential oil emulsions. The Candida krusei M4CK was isolated from the Byrsonima crassifolia fruit to make SACs. This emulsification activity (E24) was equal to or greater 50% in all carbon sources, such as olive oil, sunflower oil, kerosene, hexane, and hexadecane. E24 followed exponential growth according to the growth phase. The stability of emulsions was maintained over a wide range of temperatures, pH, and salinity. The OMBE4CK (melaleuca essential oil emulsion) had better and more significant inhibitory potential for biofilm reduction formation. In addition, bioemulsifier BE4CK alone on Escherichia coli and Pseudomonas aeruginosa biofilm showed few effective results, while there was a significant eradication for Staphylococcus aureus biofilms. The biofilms formed by S. aureus were eradicated in all concentrations of OMBE4CK. At the same time, the preformed biofilm by E. coli and P. aeruginosa were removed entirely at concentrations of 25 mg/mL, 12.5 mg/mL, and 6.25 mg/mL. The results show that the bioemulsifier BE4CK may represent a new potential for antibiofilm application.
ABSTRACT
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.
ABSTRACT
In this study, the antifungal, biosurfactant and bioemulsifying activity of the lipopeptides produced by the marine bacterium Bacillus subtilis subsp. spizizenii MC6B-22 is presented. The kinetics showed that at 84 h, the highest yield of lipopeptides (556 mg/mL) with antifungal, biosurfactant, bioemulsifying and hemolytic activity was detected, finding a relationship with the sporulation of the bacteria. Based on the hemolytic activity, bio-guided purification methods were used to obtain the lipopeptide. By TLC, HPLC and MALDI-TOF, the mycosubtilin was identified as the main lipopeptide, and it was further confirmed by NRPS gene clusters prediction based on the strain's genome sequence, in addition to other genes related to antimicrobial activity. The lipopeptide showed a broad-spectrum activity against ten phytopathogens of tropical crops at a minimum inhibitory concentration of 400 to 25 µg/mL and with a fungicidal mode of action. In addition, it exhibited that biosurfactant and bioemulsifying activities remain stable over a wide range of salinity and pH and it can emulsify different hydrophobic substrates. These results demonstrate the potential of the MC6B-22 strain as a biocontrol agent for agriculture and its application in bioremediation and other biotechnological fields.
ABSTRACT
Surface-active compounds (SACs), biomolecules produced by bacteria, yeasts, and filamentous fungi, have interesting properties, such as the ability to interact with surfaces as well as hydrophobic or hydrophilic interfaces. Because of their advantages over other compounds, such as biodegradability, low toxicity, antimicrobial, and healing properties, SACs are attractive targets for research in various applications in medicine. As a result, a growing number of properties related to SAC production have been the subject of scientific research during the past decade, searching for potential future applications in biomedical, pharmaceutical, and therapeutic fields. This review aims to provide a comprehensive understanding of the potential of biosurfactants and emulsifiers as antimicrobials, modulators of virulence factors, anticancer agents, and wound healing agents in the field of biotechnology and biomedicine, to meet the increasing demand for safer medical and pharmacological therapies.
ABSTRACT
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 24 h is the optimal time for the production of bioemulsifier by strain S3 with yield of 1.4 g/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 0.0096 h-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 (ß) 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.
Subject(s)
Bacteria/isolation & purification , Bacteria/metabolism , Emulsifying Agents/metabolism , Hot Temperature , Metabolic Networks and Pathways , Bacteria/growth & development , Biodegradation, Environmental , Hot Springs/microbiology , Hydrogen-Ion Concentration , India , KineticsABSTRACT
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.
Subject(s)
Saccharum , Trichosporon , Ammonium Hydroxide , Fermentation , Hydrolysis , Sodium HydroxideABSTRACT
In the present work, the production of bioemulsifier (BE) by a lactic acid bacterium (LAB) grown at 25⯰C in lactic whey-based media for 24â¯h was evaluated. Maximum production was detected in a medium containing yeast extract, peptone and lactic whey (LAPLW medium), with a yield of 270â¯mgâ¯L-1. The BE proved to be more innocuous for Caco-2â¯cells, used as a toxicological indicator, than the non-ionic surfactant Triton X-100. In addition, the microbial product presented higher stability to changes in temperature (37⯰C to 100⯰C), pH (2-10), and salt concentration (5% and 20%, w/v) than the synthetic surfactant. Regarding emulsifying capacity tested against different hydrophobic substrates (kerosene, motor oil, diesel, sunflower oil, and grape oil), the BE displayed E24 values similar to or even better than those of Triton X-100. Finally, Triton X-100 caused irreversible modifications on the giant unilamellar vesicles (used as model membrane system), promoting the solubilization of the lipid bilayers. Nevertheless, BE induced temporary modifications of the membrane, which is associated with incorporation of the bioproduct in the outer layer. These results demonstrate the role of BE in biological processes, including reversible changes in microbial membranes to enhance the access to hydrophobic substrates.
Subject(s)
Biotechnology/methods , Emulsifying Agents/isolation & purification , Enterococcus/metabolism , Lactic Acid/metabolism , Whey/metabolism , Caco-2 Cells , Cell Survival/drug effects , Emulsifying Agents/metabolism , Emulsifying Agents/toxicity , Emulsions , Humans , Hydrophobic and Hydrophilic Interactions , Octoxynol/chemistry , Petroleum/metabolism , Plant Oils/metabolism , TemperatureABSTRACT
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 ± 101 kDa). The best carbon source and energy for bioemulsifier production was wasted cooking oil, with a highest emulsifying capacity (76.2 ± 3.5 EU mg-1) as compared with ethanol (46.6 ± 7.1 EU mg-1) and the glycerol-hexadecane mixture (49.5 ± 4.2 EU mg-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 (0-50 g NaCl L-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.
Subject(s)
Acinetobacter/growth & development , Alkanes/pharmacology , Culture Media/pharmacology , Emulsifying Agents/metabolism , Ethanol/pharmacology , Glycerol/pharmacology , Alkanes/chemistry , Culture Media/chemistry , Ethanol/chemistry , Glycerol/chemistryABSTRACT
Bioemulsifiers are able to stabilize oil-in-water emulsions and are very important in several industrial processes, including food processing. In this study, a bioemulsifier produced by Ensifer adhaerens JHT2 was tested for its ability to emulsify edible oils (canola, corn, palm, olive and soy). Emulsification of soy and canola oils was detected, but the highest emulsification index (EI) was obtained when JHT2 culture supernatant was used for the emulsification of palm oil (EI=100%). Bioemulsifier production was evaluated using nine culture media and different NaCl concentrations (0.5 to 10%), pH (4 to 10) and temperatures (28 to 42°C). The highest emulsification activity was detected in the supernatants of JHT2 grown in trypticase soy broth containing 0.5-1.0% NaCl, pH6-7 and temperatures of 28-37°C. Characterization of the bioemulsifier produced by JHT2 revealed typical characteristics of exopolysaccharides (EPS), constituting a backbone of (1â4)-ß-d-glucopyranosyl and (1â3)-ß-D-galactopyranosyl alternating with (1â4)-α-d-mannopyranosyl units that branch from the structure at O-2. Side chains are composed of units of (1â6)-ß-d-glucopyranosyl and 3-O-linked galactopyranosyl bearing a pyruvic acid acetal substitution at O-4 and O-6. Our results indicate that the EPS produced by Ensifer adhaerens JHT2 is a promising option for improving and maintaining stable emulsions in food prepared with edible oils.
Subject(s)
Emulsifying Agents/chemistry , Plant Oils/chemistry , Polysaccharides, Bacterial/chemistry , Rhizobiaceae/chemistryABSTRACT
The objective of this study was to evaluate the capability of a lipopolysaccharide, produced by Trichosporon mycotoxinivorans CLA2 using residue of biodiesel processing, to flocculate two different solids in suspension. In addition, the emulsifying activity and the stability of this lipopolysaccharide in response to pH and temperature variations and in the presence of some electrolytes were evaluated. The lipopolysaccharide was used in concentrations ranging from 20 to 80 mgL-1 to flocculate 100 gL-1 of kaolin and 50 gL-1 of charcoal. The results indicated that the flocculating capability for each suspended particles reached 80 % and 78.79 % after 14 min, respectively. Other tests indicated that the emulsifying activity is weakly affected by temperature, pH and NaCl. In addition, the surfactant activity was assessed by the droplet diameter method and tension surface measurement. The surface tension of pure water decreased gradually with an increase in the biopolymer concentration until a minimum of 52 m Nm-1 with a CMC value of 4.54 mgL-1. These findings demonstrated a potential use of the bioemulsifier in flocculation and emulsification processes, which are also favored by its reduced toxicity compared to those of widely used commercial polymers.
Subject(s)
Emulsifying Agents/chemistry , Lipopolysaccharides/chemistry , Trichosporon/metabolism , Biofuels/analysis , Charcoal/chemistry , Emulsifying Agents/metabolism , Flocculation , Hydrogen-Ion Concentration , Kaolin/chemistry , Kinetics , Lipopolysaccharides/biosynthesis , Sodium Chloride/chemistry , Surface Tension , Temperature , Water/chemistryABSTRACT
Almost all oleaginous microorganisms are available for biodiesel production, and for the mechanism of oil accumulation, which is what makes a microbial approach economically competitive. This study investigated the potential that the yeast Candida lipolytica UCP0988, in an anamorphous state, has to produce simultaneously a bioemulsifier and to accumulate lipids using inexpensive and alternative substrates. Cultivation was carried out using waste soybean oil and corn steep liquor in accordance with 2² experimental designs with 1% inoculums (107 cells/mL). The bioemulsifier was produced in the cell-free metabolic liquid in the late exponential phase (96 h), at Assay 4 (corn steep liquor 5% and waste soybean oil 8%), with 6.704 UEA, IE24 of 96.66%, and showed an anionic profile. The emulsion formed consisted of compact small and stable droplets (size 0.2-5 µm), stable at all temperatures, at pH 2 and 4, and 2% salinity, and showed an ability to remove 93.74% of diesel oil from sand. The displacement oil (ODA) showed 45.34 cm² of dispersion (central point of the factorial design). The biomass obtained from Assay 4 was able to accumulate lipids of 0.425 g/g biomass (corresponding to 42.5%), which consisted of Palmitic acid (28.4%), Stearic acid (7.7%), Oleic acid (42.8%), Linoleic acid (19.0%), and γ-Linolenic acid (2.1%). The results showed the ability of C. lipopytica to produce both bioemulsifier and biodiesel using the metabolic conversion of waste soybean oil and corn steep liquor, which are economic renewable sources.
Subject(s)
Biofuels , Candida/metabolism , Emulsifying Agents/metabolism , Oils, Volatile/metabolism , Soybean Oil/metabolism , Biomass , Candida/growth & development , Hydrogen-Ion Concentration , Industrial Waste , Oils, Volatile/chemistry , Oleic Acid/metabolism , Palmitic Acid/metabolism , Soybean Oil/chemistry , Stearic Acids/metabolism , Substrate Specificity , Temperature , Zea mays/chemistry , Zea mays/metabolismABSTRACT
Chlordane bioremediation using actinobacteria mixed culture is an attractive clean-up technique. Their ability to produce bioemulsifiers could increase the bioavailability of this pesticide. In order to select a defined actinobacteria mixed culture for chlordane remediation, compatibility assays were performed among six Streptomyces strains. The strains did not show growth inhibition, and they were assayed for chlordane removal, either as pure or as mixed cultures. In pure cultures, all of the strains showed specific dechlorination activity (1.42-24.20 EU mg(-1)) and chlordane removal abilities (91.3-95.5%). The specific dechlorination activity was mainly improved with cultures of three or four microorganisms. The mixed culture consisting of Streptomyces sp. A2-A5-A13 was selected. Their ability to produce bioemulsifiers in the presence of glucose or chlordane was tested, but no significant differences were observed (p > 0.05). However, the stability of the emulsions formed was linked to the carbon source used. Only in chlordane presence the emulsions retained 100% of their initial height. Finally, the selected consortium showed a high degree of sporulation in the pesticide presence. This is the first study on the effects that chlordane exerts on microbe morphology and emulsifier production for a defined mixed culture of Streptomyces with ability to remediate the pesticide.
Subject(s)
Chlordan/metabolism , Emulsifying Agents/metabolism , Microbial Consortia , Pesticides/metabolism , Selection, Genetic , Streptomyces/metabolism , Biotransformation , Glucose/metabolism , Spores, Bacterial/growth & development , Streptomyces/growth & development , Streptomyces/isolation & purificationABSTRACT
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.
Subject(s)
Burkholderia/metabolism , Emulsions , Glucose/metabolism , Polyhydroxyalkanoates/metabolism , Pseudomonas/metabolism , Biomass , Burkholderia/growth & development , Pseudomonas/growth & developmentABSTRACT
Two methods were used to make crude preparations of surface-active compounds (SACs) produced by Gordonia amicalis grown on the medium containing 1% diesel oil. Using a 2:1 (v/v) solution of chloroform:methanol for extraction, Type I SACs were isolated and shown to produce oil in water (O/W) emulsions. Type II SACs were isolated by precipitation with ammonium sulfate and produced predominantly water in oil emulsions (W/O). The crude Type I and II preparations were able to produce a significant reduction in the surface tension of water; however, the crude Type II preparation had 10-25 fold higher emulsification activity than the Type I preparation. Both SAC preparations were analyzed by the TLC and each produced two distinct bands with Rf 0.44 and 0.62 and Rf 0.52 and 0.62, respectively. The partially purified SACs were characterized by the ESI(+)-MS, FT-IR and NMR. In each one of these fractions, a mixture of 10 oligomers was found consisting of a series of compounds, with masses from 502 to 899, differing in molecular mass by a repeating unit of 44 Daltons. The mass spectra of these compounds did not appear to match other known biosurfactants and could represent a novel class of these compounds.
ABSTRACT
A biomassa de levedura resultante da produção de cerveja é mátéria-prima para extração de componentes celulares, incluíndo manoproteínas. O presente trabalho avaliou a possibilidade da utilização da levedura Saccharomyces sp. descartada em cervejaria, para obtenção de extrato com manoproteínas. A extração foi conduzida segundo delineamento fatorial incompleto, Box-Behnken 33, para as variáveis temperaturas (75, 85 e 95ºC), tempo de extração (5, 7 e 9h) e concentração da suspensão de parede celular (10, 15 e 20%). O etanol residual da fermentação não interfere na obtenção do extrato contendo manoproteínas. O maior índice de extração foi 4,08%, observado para temperatura de 95ºC na concentração de 10% por 7h e 15% por 9h. A validação experimental do maior índice predito resultou em 4,50% de extrato, confirmando a capacidade preditiva do modelo. A manoproteína obtida, a partir de 10% de parede celular (95ºC, 9h), apresentou 51,39% de proteínas, com 58 e 64 kDa, e 25,89% de carboidratos, distribuídos entre manose e glicose. A atividade emulsificante foi de 62,50 ± 0,88% e a estabilidade da emulsão foi de 96,00 ± 1,40%. Estes resultados evidenciam o potencial bioemulsificante do extrato e a viabilidade de utilização da levedura descartada em cervejarias para obtenção de compostos com propriedades industriais interessantes.
The biomass of yeast after beer production is a raw-material for cell components extraction, including mannoproteins. The present study evaluated the using viability of spent brewer's yeast Saccharomyces sp. for obtainment of extract containing mannoprotein. The extraction was conducted by Box-Behnken 33 incomplete design, for the variables temperature (75, 85 and 95ºC), time of extraction (5, 7 and 9h) and concentration of cell wall in suspension (10, 15 and 20%). The residual ethanol of fermentation doesn't have interference in the obtaining of extract containing mannoproteins. The highest rate of extraction was 4.08%, obtained at 95ºC, with 10% cell wall by 7h and with 15% of cell wall during 9h. The experimental validation for obtaining of the maximum predicted resulted in 4.50% of extract, confirming the model predictable capacity. The extract containing mannoprotein obtained from 10% of cell wall (95ºC, 9h) had 51.39% of proteins, with 58 and 64 kDa, and 25.89% of carbohydrates, distributed in mannose and glucose. The emulsification activity was 62.50 ± 0.88% and the emulsion stability was 96.00 ± 1.4%. These results evidence the bioemulsifier potential of the extract and the viability of using spent yeast from brewery for obtainment of compounds with industrial interesting properties.
Subject(s)
Saccharomyces cerevisiaeABSTRACT
The biomass of yeast after beer production is a raw-material for cell components extraction, including mannoproteins. The present study evaluated the using viability of spent brewers yeast Saccharomyces sp. for obtainment of extract containing mannoprotein. The extraction was conducted by Box-Behnken 33 incomplete design, for the variables temperature (75, 85 and 95ºC), time of extraction (5, 7 and 9h) and concentration of cell wall in suspension (10, 15 and 20%). The residual ethanol of fermentation doesnt have interference in the obtaining of extract containing mannoproteins. The highest rate of extraction was 4.08%, obtained at 95ºC, with 10% cell wall by 7h and with 15% of cell wall during 9h. The experimental validation for obtaining of the maximum predicted resulted in 4.50% of extract, confirming the model predictable capacity. The extract containing mannoprotein obtained from 10% of cell wall (95ºC, 9h) had 51.39% of proteins, with 58 and 64 kDa, and 25.89% of carbohydrates, distributed in mannose and glucose. The emulsification activity was 62.50 ± 0.88% and the emulsion stability was 96.00 ± 1.4%. These results evidence the bioemulsifier potential of the extract and the viability of using spent yeast from brewery for obtainment of compounds with industrial interesting properties.
A biomass de levedura resultante da produção de cerveja é mátéria-prima para extração de componentes celulares, incluíndo manoproteínas. O presente trabalho avaliou a possibilidade da utilização da levedura Saccharomyces sp. descartada em cervejaria, para obtenção de extrato com manoproteínas. A extração foi conduzida segundo delineamento fatorial incompleto, Box-Behnken 33, para as variáveis temperaturas (75, 85 e 95ºC), tempo de extração (5, 7 e 9h) e concentração da suspensão de parede celular (10, 15 e 20%). O etanol residual da fermentação não interfere na obtenção do extrato contendo manoproteínas. O maior índice de extração foi 4,08%, observado para temperatura de 95ºC na concentração de 10% por 7h e 15% por 9h. A validação experimental do maior índice predito resultou em 4,50% de extrato, confirmando a capacidade preditiva do modelo. A manoproteína obtida, a partir de 10% de parede celular (95ºC, 9h), apresentou 51,39% de proteínas, com 58 e 64 kDa, e 25,89% de carboidratos, distribuídos entre manose e glicose. A atividade emulsificante foi de 62,50 ± 0,88% e a estabilidade da emulsão foi de 96,00 ± 1,40%. Estes resultados evidenciam o potencial bioemulsificante do extrato e a viabilidade de utilização da levedura descartada em cervejarias para obtenção de compostos com propriedades industriais interessantes