Alternative method for rhamnolipids quantification using an electrochemical platform based on reduced graphene oxide, manganese nanoparticles and molecularly imprinted Poly(L-Ser).

Rhamnolipids (RHLs) are promising biosurfactants with important applications in several industrial segments. These compounds are produced through biotechnological processes using the bacteria Pseudomonas Aeruginosa. The main methods of analyzing this compound are based on chromatographic techniques. In this study, an electrochemical sensor based on a platform modified with reduced graphene oxide, manganese nanoparticles covered with a molecularly imprinted poly (L-Ser) film was used as an alternative method to quantify RHL through its hydrolysis product, acid 3-hydroxydecanoic acid (3-HDA). The proposed sensor was characterized microscopically, spectroscopically and electrochemically. Under optimized experimental conditions, an analytical curve was obtained in the linear concentration range from 2.0 × 10-12 mol L-1 to 1.0 × 10-10 mol L-1. The values estimated of LOD, LOQ and AS were 8.3 × 10-13 mol L-1, 2.7 × 10-12 mol L-1and 1.3 × 107 A L mol-1, respectively. GCE/rGO/MnNPs/L-Ser@MIP exhibits excellent selectivity, repeatability, and high stability for the detection of 3-HDA. Furthermore, the developed method was successfully applied to the recognition of the hydrolysis product (3-HDA) of RHLs obtained from guava agro-waste. Statistical comparison between GCE/rGO/MnNPs/L-Ser@MIP and HPLC method confirms the accuracy of the electrochemical sensor within a 95% confidence interval.

Rhamnolipids as Green Stabilizers of nZVI and Application in the Removal of Nitrate From Simulated Groundwater.

Environmental contamination caused by inorganic compounds is a major problem affecting soils and surface water. Most remediation techniques are costly and generally lead to incomplete removal and production of secondary waste. Nanotechnology, in this scenario with the zero-valent iron nanoparticle, represents a new generation of environmental remediation technologies. It is non-toxic, abundant, cheap, easy to produce, and its production process is simple. However, in order to decrease the aggregation tendency, the zero-iron nanoparticle is frequently coated with chemical surfactants synthesized from petrochemical sources, which are persistent or partially biodegradable. Biosurfactants (rhamnolipids), extracellular compounds produced by microorganisms from hydrophilic and hydrophobic substrates can replace synthetic surfactants. This study investigated the efficiency of a rhamnolipid biosurfactant on the aggregation of nanoscale zer-valent iron (nZVI) and its efficiency in reducing nitrate in simulated groundwater at pH 4.0. Two methods were tested: 1) adding the rhamnolipid during chemical synthesis and 2) adding the rhamnolipid after chemical synthesis of nZVI. Scanning electron microscopy field emission, X-ray diffractometry, Fourier transform infrared spectroscopy, thermogravimetric analysis, Dynamic Light Scattering, and zeta potential measurements were used to characterize bare nZVI and rhamnolipid-coated nZVI. The effects of the type of nZVI and initial NO3 concentration were examined. Nanoscale zer-valent iron with the addition of the rhamnolipid after synthesis achieved the best removal rate of nitrate (about 78%), with an initial nitrate concentration of 25 mg L-1. The results suggest that nZVI functionalized with rhamnolipids is a promising strategy for the in situ remediations of groundwater contaminated by NO3, heavy metal, and inorganic carbon.

Rhamnolipids and essential oils in the control of mosquito-borne tropical diseases.

The diseases transmitted by mosquito vectors are a great public health issue. Thus, effective vector control becomes the main strategy to reduce their prevalence. However, insecticide resistance has become a huge concern for the mitigation of mosquitoes; here, we propose the use of rhamnolipids in emulsion with clove oil against Aedes aegypti and Culex quinquefasciatus. The toxicity of rhamnolipids and clove oil to two species of mosquitoes transmitting tropical diseases was investigated. After 24 h, the LC50 was 140 mg/L when rhamnolipids were used and 154 mg/L when clove oil was used against Aedes aegypti larvae. In the case of Culex quinquefasciatus, the LC50 was 130 mg/L for rhamnolipids and 19 mg/L for clove oil. When the concentrations of the upper limits of one of the solutions (rhamnolipid or clove oil) were mixed, 100% mortality was obtained after 24 h. The bioassay of insecticidal action for solutions of rhamnolipids and clove oil in the lower limit, upper limit, and lethal concentration 50 to determine the effect on 50% of the population (KD50) achieved low results from KD50 to the upper limit compared to the other concentrations for both Aedes aegypti and Culex quinquefasciatus. The rhamnolipids and clove oil at the upper limit concentration had the greatest repellent activity against the two mosquito species. Bioassays using different concentrations of rhamnolipids revealed variations in the morphology of the intestinal epithelium (800 mg/L). A concentration of 900 mg/L led to the most severe morphological changes in the organization of the epithelium and the cells lining the intestines of these larvae. When larvae were exposed to a concentration of 1000 mg/L, the marginalization of chromatin in the nucleus of epithelial cells was very severe, indicating the onset of cell death.Key points• The toxicity of rhamnolipids and clove oil has a larvicidal, insecticidal, and repellent effect.• The combination of concentrations of these compounds enhances their action.• Different concentrations of rhamnolipids led to severe morphological changes in the organization of the epithelium and the cells and the intestines of larvae.

Silver nanoparticles stabilized by ramnolipids: Effect of pH.

Rhamnolipids are glycolipid biosurfactants that have remarkable physicochemical characteristics, such as the capacity for self-assembly, which makes these biomolecules a promising option for application in nanobiotechnology. Rhamnolipids produced from a low-cost carbon source (glycerol) were used to stabilize silver nanoparticles. Silver nanoparticles (AgNPs) have been the subject of studies due to their physical chemical as well as biological properties, which corroborate their catalytic and antimicrobial activity. We compared nanoparticles obtained with three different pH values during synthesis (5, 7 and 9) in the presence of rhamnolipids. Dynamic light scattering showed that larger particles were formed at pH 5 (78-190 nm) compared to pH 7 (6.5-43 nm) and 9 (5.6-28.1 nm). Moreover, nanoparticle stability (analyzed based on the zeta potential) was enhanced with the increase in pH from 5 to 9 (-29.86 ± 1.04, -37.83 ± 0.90 and -40.33 ± 0.57 mV, respectively). Field emission gun scanning electron microscopy confirmed the round morphology of the silver nanoparticles. The LSPR spectra of AgNP for the pHs studied are conserved. In conclusion, different pH values in the presence of rhamnolipids used in the synthesis of silver nanoparticles directly affect nanoparticle size and stability.

Rhamnolipid-Based Liposomes as Promising Nano-Carriers for Enhancing the Antibacterial Activity of Peptides Derived from Bacterial Toxin-Antitoxin Systems.

BACKGROUND: Antimicrobial resistance poses substantial risks to human health. Thus, there is an urgent need for novel antimicrobial agents, including alternative compounds, such as peptides derived from bacterial toxin-antitoxin (TA) systems. ParELC3 is a synthetic peptide derived from the ParE toxin reported to be a good inhibitor of bacterial topoisomerases and is therefore a potential antibacterial agent. However, ParELC3 is inactive against bacteria due to its inability to cross the bacterial membranes. To circumvent this limitation we prepared and used rhamnolipid-based liposomes to carry and facilitate the passage of ParELC3 through the bacterial membrane to reach its intracellular target - the topoisomerases. METHODS AND RESULTS: Small unilamellar liposome vesicles were prepared by sonication from three formulations that included 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine and cholesterol. ParELC3 was loaded with high efficiency into the liposomes. Characterization by DLS and TEM revealed the appropriate size, zeta potential, polydispersity index, and morphology. In vitro microbiological experiments showed that ParELC3 loaded-liposomes are more efficient (29 to 11 µmol·L-1) compared to the free peptide (>100 µmol·L-1) at inhibiting the growth of standard E. coli and S. aureus strains. RL liposomes showed high hemolytic activity but when prepared with POPC and Chol this activity had a significant reduction. Independently of the formulation, the vesicles had no detectable cytotoxicity to HepG2 cells, even at the highest concentrations tested (1.3 mmol·L-1 and 50 µmol·L-1 for rhamnolipid and ParELC3, respectively). CONCLUSION: The present findings suggest the potential use of rhamnolipid-based liposomes as nanocarrier systems to enhance the bioactivity of peptides.

A New Possibility for Fermentation Monitoring by Electrical Driven Sensing of Ultraviolet Light and Glucose.

Industrial fermentation generates products through microbial growth associated with the consumption of substrates. The efficiency of industrial production of high commercial value microbial products such as ethanol from glucose (GLU) is dependent on bacterial contamination. Controlling the sugar conversion into products as well as the sterility of the fermentation process are objectives to be considered here by studying GLU and ultraviolet light (UV) sensors. In this work, we present two different approaches of SnO2 nanowires grown by the Vapor-Liquid-Solid (VLS) method. In the GLU sensor, we use SnO2 nanowires as active electrodes, while for the UV sensor, a nanowire film was built for detection. The results showed a wide range of GLU sensing and as well as a significant influence of UV in the electrical signal. The effect of a wide range of GLU concentrations on the responsiveness of the sensor through current-voltage based on SnO2 nanowire films under different concentration conditions ranging was verified from 1 to 1000 mmol. UV sensors show a typical amperometric response of SnO2 nanowires under the excitation of UV and GLU in ten cycles of 300 s with 1.0 V observing a stable and reliable amperometric response. GLU and UV sensors proved to have a promising potential for detection and to control the conversion of a substrate into a product by GLU control and decontamination by UV control in industrial fermentation systems.

Efficient Conversion of Agroindustrial Waste into D(-) Lactic Acid by Lactobacillus delbrueckii Using Fed-Batch Fermentation.

PURPOSE: The goal of this paper is to describe the green conversion of agricultural waste products, such as molasses and corn steep liquor, into large amounts of D(-) lactic acid using a facilitated multipulse fed-batch strategy and affordable pH neutralizer. This is a very low-cost process because there is no need for hydrolysis of the waste products. The fed-batch strategy increases lactic acid productivity by avoiding inhibition caused by a high initial substrate concentration, and the selected controlling agent prevents cell stress that could be caused by high osmotic pressure of the culture media. METHODS: The effects of different carbon and nitrogen sources on lactic acid production were investigated, and the best concentrations of the medium components were determined. To optimize the culture conditions of the Lactobacillus delbrueckii strain, the effects of pH control, temperature, neutralizing agent, agitation, and inoculum size in batch cultures were investigated. Fed-batch strategies were also studied to improve production and productivity. RESULT: A high titer of D(-) lactic acid (162g/liter) was achieved after 48 hours of fermentation. Productivity at this point was 3.37 g/L·h. The optimum conditions were a temperature of 39°C, pH 5.5 controlled by the addition of Ca(OH)2, agitation at 150 rpm, and inoculum size of 25% (v/v). CONCLUSION: The production of high optical purity D(-) lactic acid through L. delbrueckii fermentation with molasses and corn steep liquor is a promising economical alternative process that can be performed on the industrial scale.

Polyhydroxyalkanoate Synthesis by Burkholderia glumae into a Sustainable Sugarcane Biorefinery Concept.

Polyhydroxyalkanoate (PHA) bioplastic was synthesized by Burkholderia glumae MA13 from carbon sources and industrial byproducts related to sugarcane biorefineries: sucrose, xylose, molasses, vinasse, bagasse hydrolysate, yeast extract, yeast autolysate, and inactivated dry yeast besides different inorganic nitrogen sources. Sugarcane molasses free of pre-treatment was the best carbon source, even compared to pure sucrose, with intracellular polymer accumulation values of 41.1-46.6% cell dry weight. Whereas, xylose and bagasse hydrolysate were poor inducers of microbial growth and polymer synthesis, the addition of 25% (v/v) sugarcane vinasse to the culture media containing molasses was not deleterious and resulted in a statistically similar maximum polymer content of 44.8% and a maximum PHA yield of 0.18 g/g, at 34°C and initial pH of 6.5, which is economic and ecologically interesting to save water required for the industrial processes and especially to offer a fermentative recycling for this final byproduct from bioethanol industry, as an alternative to its inappropriate disposal in water bodies and soil contamination. Ammonium sulfate was better even than tested organic nitrogen sources to trigger the PHA synthesis with polymer content ranging from 29.7 to 44.8%. GC-MS analysis showed a biopolymer constituted mainly of poly(3-hydroxybutyrate) although low fractions of 3-hydroxyvalerate monomer were achieved, which were not higher than 1.5 mol% free of copolymer precursors. B. glumae MA13 has been demonstrated to be adapted to synthesize bioplastics from different sugarcane feedstocks and corroborates to support a biorefinery concept with value-added green chemicals for the sugarcane productive chain with additional ecologic benefits into a sustainable model.

Active-electrode biosensor of SnO2 nanowire for cyclodextrin detection from microbial enzyme.

Cyclodextrin (CD) is a conical compound used in food and pharmaceutical industry to complexation of hydrophobic substances. It is a product of microbial enzymes which converts starch into CD during their activity. We aim to detect CD using active-electrode biosensor of SnO2. They were grown on active electrode by the VLS method. The CD consists of several glucose units which have hydroxyl groups which tend to bind to interface states present in nanowires changing their conductivity. Experimental results of electrical conductivity at different CD concentrations are presented. A model that describes the influence of adsorbed glucose on nanowires and its electrical properties is also presented. Some general observations are performed on the applicability of the CD adsorption method by the nanowire-based biosensor.

High-titer and productivity of l-(+)-lactic acid using exponential fed-batch fermentation with Bacillus coagulans arr4, a new thermotolerant bacterial strain.

Bacillus coagulans arr4 is a thermotolerant microorganism with great biotechnological potential for l-(+)-lactic acid production from granulated sugar and yeast extract. The highest l-(+)-lactic acid production was obtained with Ca(OH)2. The maximum production of l-(+)-lactic acid (206.81 g/L) was observed in exponential feeding using granulated sugar solution (900 g/L) and yeast extract (1%) at 50 °C, pH 6.5, and initial granulated sugar concentration of 100 g/L at 39 h. 5.3 g/L h productivity and 97% yield were observed, and no sugar remained. Comparing the simple batch with exponential fed-batch fermentation, the l(+) lactic acid production was improved in 133.22% and dry cell weight was improved in 83.29%, using granulated sugar and yeast extract. This study presents the highest productivity of lactic acid ever observed in the literature, on the fermentation of thermotolerant Bacillus sp. as well as an innovative and high-efficiency purification technology, using low-cost substances as Celite and charcoal. The recovery of lactic acid was 86%, with 100% protein removal, and the fermentation medium (brown color) became a colorless solution.

Biosurfactants produced by Scheffersomyces stipitis cultured in sugarcane bagasse hydrolysate as new green larvicides for the control of Aedes aegypti, a vector of neglected tropical diseases.

Biosurfactants are microbial metabolites with possible applications in various industrial sectors that are considered ecofriendly molecules. In recent years, some studies identified these compounds as alternatives for the elimination of vectors of tropical diseases, such as Aedes aegypti. The major bottlenecks of biosurfactant industrial production have been the use of conventional raw materials that increase production costs as well as opportunistic or pathogenic bacteria, which restrict the application of these biomolecules. The present study shows the potential of hemicellulosic sugarcane bagasse hydrolysate as a raw material for the production of a crystalline glycolipidic BS by Scheffersomyces stipitis NRRL Y-7124, which resulted in an emulsifying index (EI24) of 70 ± 3.4% and a superficial tension of 52 ± 2.9 mN.m-1. Additionally, a possible new application of these compounds as biolarvicides, mainly against A. aegypti, was evaluated. At a concentration of 800 mg.L-1, the produced biosurfactant caused destruction to the larval exoskeletons 12 h after application and presented an letal concentration (LC50) of 660 mg.L-1. Thus, a new alternative for biosurfactant production using vegetal biomass as raw material within the concept of biorefineries was proposed, and the potential of the crystalline glycolipidic biosurfactant in larvicidal formulations against neglected tropical disease vectors was demonstrated.

Environmentally Friendly Production of D(-) Lactic Acid by Sporolactobacillus nakayamae: Investigation of Fermentation Parameters and Fed-Batch Strategies.

The interest in the production of lactic acid has increased due to its wide range of applications. In the present study, the variables that affect fermentative D(-) lactic acid production were investigated: neutralizing agents, pH, temperature, inoculum percentage, agitation, and concentration of the medium components. An experimental design was applied to determine the optimal concentrations of the medium components and fermentation was studied using different feeding strategies. High production (122.41 g/L) and productivity (3.65 g/L·h) were efficiently achieved by Sporolactobacillus nakayamae in 54 h using a multipulse fed-batch technique with an initial medium containing 35 g/L of yeast extract (byproduct of alcohol production), 60 g/L of crystallized sugar, and 7.5 mL/L of salts. The fermentation process was conducted at 35°C and pH 6.0 controlled by NaOH with a 20% volume of inoculum and agitation at 125 rpm. The production of a high optically pure concentration of D(-) lactic acid combined with an environmentally friendly NaOH-based process demonstrates that S. nakayamae is a promising strain for D(-) lactic acid production.

Biodiesel byproduct bioconversion to rhamnolipids: Upstream aspects.

This study focused on two important aspects of the upstream process: the appropriate use of crude glycerol as a low-cost carbon source, and strain selection. The effect of different crude glycerol concentrations on rhamnolipid biosynthesis by two Pseudomonas aeruginosa strains (wild type LBI and mutant LBI 2A1) was studied. Finally, the synthesized rhamnolipids were characterized by mass spectrometry. When both strains were compared, 50 g/L was the most favorable concentration for both, but P. aeruginosa LBI 2A1 showed an increase in rhamnolipid production (2.55 g/L) of 192% over wild type (1.3 g/L). The higher rhamnolipid production could be related to a possible mechanism developed after the mutation process at high antibiotic concentrations. Mass spectrometry confirmed the glycolipid nature of the produced biosurfactant, and the homologue composition showed a wide mixture of mono and di-rhamnolipids. These results show that high glycerol concentrations can inhibit microbial metabolism, due to osmotic stress, leading to a better understanding of glycerol metabolism towards its optimization in fermentation media. Since P. aeruginosa LBI 2A1 showed higher conversion yields than P. aeruginosa LBI, the use of a mutant strain associated with a low cost carbon source might improve biosurfactant biosynthesis, therefore yielding an important upstream improvement.

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) production from biodiesel by-product and propionic acid by mutant strains of Pandoraea sp.

Pandoraea sp. MA03 wild type strain was subjected to UV mutation to obtain mutants unable to grow on propionic acid (PA) but still able to produce poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] from glycerol and PA at high 3HV yields. In shake flask experiments, mutant prp25 was selected from 52 mutants affected in the propionate metabolism exhibiting a conversion rate of PA into 3HV units of 0.78 g g-1 . The use of crude glycerol (CG) plus PA or valeric acid resulted in a copolymer with 3HV contents varying from 21.9 to 30 mol% and 22.2 to 36.7 mol%, respectively. Fed-batch fermentations were performed using CG and PA and reached a 3HV yield of 1.16 g g-1 , which is 86% of the maximum theoretical yield. Nitrogen limitation was a key parameter for polymer accumulation reaching up to 63.7% content and 18.1 mol% of 3HV. Henceforth, mutant prp25 is revealed as an additional alternative to minimize costs and support the P(3HB-co-3HV) production from biodiesel by-products. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1077-1084, 2017.

L-(+)-Lactic acid production by Lactobacillus rhamnosus B103 from dairy industry waste

ABSTRACT Lactic acid, which can be obtained through fermentation, is an interesting compound because it can be utilized in different fields, such as in the food, pharmaceutical and chemical industries as a bio-based molecule for bio-refinery. In addition, lactic acid has recently gained more interest due to the possibility of manufacturing poly(lactic acid), a green polymer that can replace petroleum-derived plastics and be applied in medicine for the regeneration of tissues and in sutures, repairs and implants. One of the great advantages of fermentation is the possibility of using agribusiness wastes to obtain optically pure lactic acid. The conventional batch process of fermentation has some disadvantages such as inhibition by the substrate or the final product. To avoid these problems, this study was focused on improving the production of lactic acid through different feeding strategies using whey, a residue of agribusiness. The downstream process is a significant bottleneck because cost-effective methods of producing high-purity lactic acid are lacking. Thus, the investigation of different methods for the purification of lactic acid was one of the aims of this work. The pH-stat strategy showed the maximum production of lactic acid of 143.7 g/L. Following purification of the lactic acid sample, recovery of reducing sugars and protein and color removal were 0.28%, 100% and 100%, respectively.

Agave syrup as a substrate for inulinase production by Kluyveromyces marxianus NRRL Y-7571 / Xarope de agave como substrato para produção de inulinase por Kluyveromyces marxianus NRRL Y-7571

The factorial planning was used to plan and optimize inulinase production by the yeast Kluyveromyces marxianus NRRL Y-7571. The experiments were conducted using a Central Composite Design (CCD) 22, at different concentrations of agave syrup (3.6 to 6.4%) and yeast extract (2.2 to 3.0%). After 96 hours of fermentation, the best condition for the inulinase production was 5% agave syrup and 2.5% yeast extract, which yielded an average of 129.21 U mL-1 of inulinase. Partial characterization of the crude enzyme showed that the optimal pH and temperature were 4.0 and 60°C, respectively. The enzyme showed thermal stability at 55°C for 4 hours.

O planejamento fatorial foi utilizado para planejar e otimizar a produção de inulinase pela levedura Kluyveromyces marxianus NRRL Y-7571. Os experimentos foram conduzidos por meio de Delineamento Composto Central Rotacional (DCCR) 22 em diferentes concentrações de xarope de agave (3,6 a 6,4%) e extrato de levedura (2,2 a 3,0%). Depois de 96 horas de fermentação, a melhor condição para produção de inulinase foi xarope de agave 5% e extrato de levedura 2,5%, com uma produção média de 129,21 U mL-1. A caracterização parcial do extrato enzimático bruto mostrou que a enzima apresenta pH e temperatura ótimos de 4,0 e 60oC, respectivamente. A enzima mostrou estabilidade térmica a 55oC durante 4 horas.

l-(+)-Lactic acid production by Lactobacillus rhamnosus B103 from dairy industry waste.

Lactic acid, which can be obtained through fermentation, is an interesting compound because it can be utilized in different fields, such as in the food, pharmaceutical and chemical industries as a bio-based molecule for bio-refinery. In addition, lactic acid has recently gained more interest due to the possibility of manufacturing poly(lactic acid), a green polymer that can replace petroleum-derived plastics and be applied in medicine for the regeneration of tissues and in sutures, repairs and implants. One of the great advantages of fermentation is the possibility of using agribusiness wastes to obtain optically pure lactic acid. The conventional batch process of fermentation has some disadvantages such as inhibition by the substrate or the final product. To avoid these problems, this study was focused on improving the production of lactic acid through different feeding strategies using whey, a residue of agribusiness. The downstream process is a significant bottleneck because cost-effective methods of producing high-purity lactic acid are lacking. Thus, the investigation of different methods for the purification of lactic acid was one of the aims of this work. The pH-stat strategy showed the maximum production of lactic acid of 143.7g/L. Following purification of the lactic acid sample, recovery of reducing sugars and protein and color removal were 0.28%, 100% and 100%, respectively.

Rhamnolipids: solution against Aedes aegypti?

Aedes aegypti mosquitoes are the primary transmitters of dengue fever, urban yellow fever, and chikungunya viruses. This mosquito has developed resistance to the insecticides currently used to control their populations. These chemical insecticides are harmful to the environment and can have negative effects on human health. Rhamnolipids are environmentally compatible biological surfactants, but their insecticidal activity has not been extensively studied. The present study evaluated the potential larvicidal, insecticidal, and repellent activities of rhamnolipids against A. aegypti. At concentrations of 800, 900, and 1000 mg/L, rhamnolipids eliminated all mosquito larvae in 18 h and killed 100% of adults at 1000 mg/L. According to the results it may be conclude that rhamnolipids should be applied to control larvae and mosquitos besides present the repellency activity against A. aegypti.

Produção de glicose e frutose por invertase de Saccharomyces cerevisiae imobilizada em suporte MANAE-Agarose / Glucose and Fructose Production by Saccharomyces cerevisiae Invertase Immobilized on MANAE-Agarose Support

Invertase from Saccharomyces cerevisiae was immobilized on agarose beads, activated with various groups (glyoxyl, MANAE or glutaraldehyde), and on some commercial epoxy supports (Eupergit and Sepabeads). Very active and stable invertase derivatives were produced by the adsorption of the enzyme on MANAE-agarose, MANAE-agarose treated with glutaraldhyde and glutaraldehyde-agarose supports. At pH 5.0, these derivatives retained full activity after 24h at 40 ºC and 50 ºC. When assayed at 40 °C and 50 °C, with the pH adjusted to 7.0, the invertase-MANAE-agarose derivative treated with glutaraldehyde retained 80% of the initial activity. Recovered activities of the derivatives produced with MANAE, MANAE treated with glutaraldehyde and glutaraldehyde alone were 73.5%, 44.4% and 36.8%, respectively. These three preparations were successfully employed to produce glucose and fructose in 3 cycles of sucrose hydrolysis.

Invertase de Saccharomyces cerevisiae foi imobilizada em agarose ativada com diferentes grupos (glioxil, MANAE ou glutaraldeído) e suportes epóxidos comerciais (Eupergit e Sepabeads). Derivados de invertase ativos e estabilizados foram produzidos pela adsorção da enzima em suportes MANAE-agarose, MANAE-agarose tratado com glutaraldeído e glutaraldeído-agarose. Em pH 5,0 estes derivados retiveram total atividade até 24h a 40 ºC e 50 ºC. Quando os ensaios foram a 40 °C e 50 °C com o pH alterado para 7,0, o derivado invertase-MANAE-agarose tratado com glutaraldeído apresentou 80% da atividade inicial. As atividades recuperadas dos derivados foram 73,5%, 44,4% e 36,8%, respectivamente para MANAE, MANAE tratado com glutaraldeído e glutaraldeído. Essas três preparações foram empregadas com sucesso em 3 ciclos de hidrólise da sacarose para produzir glicose e frutose.

Rhamnolipid emulsifying activity and emulsion stability: pH rules.

Rhamnolipids have excellent emulsifying power with a variety of hydrocarbons, aromatic compounds and vegetable oils. This paper evaluates the effect of pH ranging from 3 to 9 on the emulsifying activity and stability of rhamnolipids and hydrophobic substrates (benzene, soybean oil and kerosene). Sodium dodecyl sulfate was used as a reference to compare the level of chemical and biological surfactant activity after 24 h (E(24)). The results indicate that pH exerts an influence over emulsion formation and stability. For rhamnolipids, peak emulsifying activity occurred under base conditions for all substrates, with the highest value achieved with kerosene at pH 8. The results of the present study demonstrate that rhamnolipids could replace chemical surfactants, such as SDS, in different industrial fields.