45 research outputs found
Non-ionic aqueous micellar extraction of trypsin inhibitors and isoflavones from soybean meal: process optimization
Simultaneous extraction of trypsin inhibitors and soy isoflavones from soybean meal was investigated using the non-destructive phytochemical extraction process, namely aqueous micellar system. The ethoxylated aliphatic alcohols Genapol X-080, Tergitol 15-S-7, and Tergitol 15-S-9, all non-toxic and biodegradable surfactants, were assessed as potential extractants. A Box-Behnken multifactorial design with the application of the Derringer desirability was used to determine the conditions that maximized the trypsin inhibitors and isoflavone extraction while minimizing the protein extraction. The optimum condition of 5% m/m of surfactant in 50 mM aqueous sodium citrate solution pH 4.5, at 45 °C for 45 min, was established for the three surfactants. The novel methodology would allow the extraction of the main soybean antinutritional factors, trypsin inhibitors, and the valuable isoflavones, preserving the nutritional quality of the treated material. This represents a sustainable alternative methodology for industrial purposes due to its low cost, biodegradability, non-toxicity, and easy scaling up.info:eu-repo/semantics/publishedVersio
Production of soy protein concentrate with the recovery of bioactive compounds: from destruction to valorization
This work aimed to develop a novel methodology based on aqueous micellar systems (AMS), for producing soy protein concentrates (SPC) from soybean flour and recovering high-valuable bioactive compounds as by-products. Ethoxylated aliphatic alcohols Tergitol 15-S-7 and Tergitol 15-S-9, non-toxic and biodegradable surfactants, were selected to form the AMS. The methodology consisted of an extractive stage of soybean flour with AMS, which rendered both a pellet, i.e., the SPC, and a supernatant containing the extracted bioactive compounds. The latter was further heated above the cloud point temperature, thus resulting in a biphasic system formed by a micelle-rich phase (MP) and an aqueous phase (AP). Obtained SPC showed a noticeable loss (∼90%) of trypsin inhibitor activity, a total protein content close to 60%, soluble protein amounts varying from 19% to 34%, and remarkable released (by simulated digestion) antioxidant and antihypertensive activities. Those indicators are similar to or even better than those corresponding to SPC from the classical acid-extraction method. The AMS also exhibited an enhanced efficiency for extracting antinutrients such as non-digestible oligosaccharides, trypsin inhibitors, and lectins mostly recovered at the AP and separated from isoflavones, which were concentrated and isolated at the MP. The recovery of all the mentioned bioactive compounds, whether beneficial or undesirable, broadens their uses in research, food, and pharmacological fields. This successful performance, simplicity, scalability, and sustainability make the proposed AMS-based extraction a powerful tool for processing plant derivatives and valorizing their by-products.info:eu-repo/semantics/publishedVersio
Liquid–liquid equilibrium of the Ucon 50-HB5100/sodium citrate aqueous two-phase systems
The phase diagrams of Ucon 50-HB5100/sodium citrate aqueous two-phase systems were determined at 5, 20 and 40 ◦C. Two medium pHs
5.20 and 8.20 were assayed. The binodal curves were satisfactorily described using a four-parameter sigmoidal equation. The two-phase area was
expanded by increasing both pH and temperature. The reliability of the measured tie line compositions was ascertained by correlation equations
given by Othmer–Tobias and Bancroft.VALNATURACONICETALFA II-0440-FA-Exchange programme between Universities
of the European Union and Latin Americ
Liquid-liquid equilibrium and partitioning features of bovine trypsin in Ucon 50 HB5100 /sodium citrate aqueous two phase systems
The phase diagrams of Ucon 50-HB-5100, a non-ionic random copolymer of ethylene oxide and propylene
oxide (EOPO) and sodium citrate aqueous two-phase systems were determined at different pHs (5.20
and 8.20) and temperatures (5, 20 and 40º C). The binodal curves were determined by refractive index and
enzymatic assay of the solution and described using a four-parameter sigmoidal equation, the reliability
of the measured tie line compositions was ascertained by correlation equations given by Othmer Tobias
and Bancroft. The two-phase area was expanded by increasing both pH and temperature. The partitioning
of bovine trypsin and a-chymotrypsin, proteases of similar physico-chemical properties was investigated
in order to evaluate the applicability of partitioning as a putative method to isolate from pancreas and to
obtain any information about their partitioning mechanism. The effect of different factors such as pH, tie
line length and the presence of an inorganic salt on the protein partition coefficient were analyzed.Se caracterizaron las curvas binomiales correspondientes a los sistemas bifásicos acuosos formados por
Ucon 50-HB-5100, un copolímero al azar de óxido de etileno y óxido de propileno (EOPO) y citrato de
sodio a diferentes pHs (5,20 y 8,20) y temperaturas (5, 20 y 40º C). Las curvas binomiales se obtuvieron
por determinación del índice de refracción y ensayos enzimáticos de las soluciones correspondientes; las
composiciones de las líneas de unión se corroboraron por las ecuaciones propuestas por Othmer Tobias y
Bancroft. El aumento del pH y la temperatura condujeron a un aumento del área bifásica. También se
ensayó el comportamiento de reparto de dos proteasas tripsina y α-quimotripsina con el objetivo de
emplear los principios de partición como método de aislamiento y purificación de Tripsina a partir de páncreas
bovino y de obtener información acerca del mecanismo de partición de la misma. Se analizó el efecto
del pH, longitud de la línea de unión y presencia de sales inorgánicas sobre el coeficiente de partición
de las enzimas
Aqueous micellar two-phase system as an alternative method to selectively remove soy antinutritional factors
In this work, different antinutritional factors (trypsin inhibitors, isoflavones and raffinose family oligosaccharides) were selectively removed from soy flour by using aqueous micellar two-phase systems (AMTPS). The effects of independent variables including temperature (30–60 °C), time (10–40 min) and solid to liquid ratio (0.025–0.050 g/L) on the extraction of each antinutritional factor were analyzed using a full factorial design. As general tendency, temperature and time were the most significant parameters (p < 0.05). The best condition for the selective recovery (97% of isoflavones at top phase, and more than 50% of the rest of ANFs at bottom phase) were 5 g/L of Genapol X-080, 0.2 moL/L of sodium citrate pH 5.00, 30 °C, 40 min and 0.050 g/L. Besides, in vitro gastrointestinal digestions assays demonstrated that the treated soy flour improved its protein digestibility. The findings of this work represent the introduction of a novel methodology to selectively remove soy antinutritional factors.Fil: Haidar, Carla Nahir. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Coscueta, Ezequiel Ricardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Cordisco, Estefanía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Nerli, Bibiana Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Pellegrini Malpiedi, Luciana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentin
Biosurfactants produced by Pseudomonas syringae pv tabaci: A versatile mixture with interesting emulsifying properties
In recent years, worldwide awareness about environmental concerns has led to an extensive searching for products of natural origin. Biosurfactants (BS), are an attractive alternative to the chemical counterparts. In these sense, this works evaluate for the first time the emulsifying and surface activity of biosurfactants produced by P. syringae pv. tabaci. Chemical characterization revealed that the produced BS extract mostly consisted in a mixture of syringopeptins and arthrofactins. Besides, this sample was able to reduce water surface tension up to 36.89 mN/m. This activity was improved by modifying conditions of pH, temperature and salinity. For instance, the surface tension was reduced up to 29 mN/m when using NaCl 5 % w/w. The BS also exhibited a high emulsifying activity, this being comparable or even superior to that of commercial bioemulsifiers such xanthan, tara and guar gums. A notorious emulsification index, close to 90 %, was exhibited by BS in coconut oil/water mixtures. Moreover, emulsifying activity resulted to be little affected by different operational conditions, thus retaining about 80 % of its emulsificant index in presence of 20 % w/w of NaCl at 80 °C. These findings are indicative of the BS promising properties and its potential as additive in a variety of industrial applications.Fil: Haidar, Carla Nahir. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Pereira, Matheus M.. Universidade Tiradentes; BrasilFil: Lima, Álvaro S.. Universidade Tiradentes; BrasilFil: Nerli, Bibiana Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Pellegrini Malpiedi, Luciana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentin
Cloud point extraction based on non-ionic surfactants: An ecofriendly tool for recovering papain from papaya latex
The potential use of cloud point extraction with aqueous micellar two-phase systems (AMTPS), formed by the non-ionic biodegradable surfactants Triton X-114 (TX-114) and Genapol X-080 (GX-080), to recover papain (PAP) from fresh papaya latex was evaluated. Binodal curves obtained for both surfactants, in absence and presence of PAP, showed cloud points below 40 °C, the PAP presence slightly favoring the phase separation phenomenon. Partitioning experiments showed that PAP migrated toward the micelle-poor phase independently of the surfactant and the working temperature assayed, this behavior being consistent with this enzyme hydrophilic character. PAP recoveries of over 60% in the poor-micelle phase were obtained when partitioned fresh papaya latex in both surfactant-based AMTPS. TX-114 systems (4.00% w/w, 33 °C) allowed recovering 66% of PAP with a purification factor of 4.6. On GX-080 systems (4.00% w/w, 42 °C) a yield of 78% PAP and a purification factor of 3.7 were achieved. This AMTPS also allowed the separation of PAP from reducing sugars present in papaya latex. These results show that TX-114 and GX-080-based AMTPS could be potentially used as simple and economical tools at recovering PAP from papaya latex.Fil: DI GIACOMO, MELISA. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Bertoni, Fernando Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Rocha, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Nerli, Bibiana Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Rodriguez, Fernanda Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentin
An alternative method to isolate protease and phospholipase A2 toxins from snake venoms based on partitioning of aqueous two-phase systems
Snake venoms are rich sources of active proteins that have been employed in the diagnosis and treatment of health disorders and antivenom therapy. Developing countries demand fast economical downstream processes for the purification of this biomolecule type without requiring sophisticated equipment. We developed an alternative, simple and easy to scale-up method, able to purify simultaneously protease and phospholipase A2 toxins from Bothrops alternatus venom. It comprises a multiple-step partition procedure with polyethylene-glycol/phosphate aqueous two-phase systems followed by a gel filtration chromatographic step. Two single bands in SDS-polyacrylamide gel electrophoresis and increased proteolytic and phospholipase A2 specific activities evidence the homogeneity of the isolated proteins.Fil: Gomez, Gabriela Noemi. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; ArgentinaFil: Nerli, Bibiana Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Acosta, Ofelia Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentina. Universidad Nacional del Nordeste; ArgentinaFil: Picó, Guillermo Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Leiva, Laura Cristina Ana. Universidad Nacional del Nordeste; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentin
Molecular features determining different partitioning patterns of papain and bromelain in aqueous two-phase systems
The partitioning patterns of papain (PAP) and bromelain (BR), two well-known cysteine-proteases, in polyethyleneglycol/sodium citrate aqueous two-phase systems (ATPSs) were determined. Polyethyleneglycols of different molecular weight (600, 1000, 2000, 4600 and 8000) were assayed. Thermodynamic characterization of partitioning process, spectroscopy measurements and computational calculations of protein surface properties were also carried out in order to explain their differential partitioning behavior. PAP was observed to be displaced to the salt-enriched phase in all the assayed systems with partition coefficients (KpPAP) values between 0.2 and 0.9, while BR exhibited a high affinity for the polymer phase in systems formed by PEGs of low molecular weight (600 and 1000) with partition coefficients (KpBR) values close to 3. KpBR values resulted higher than KpPAP in all the cases. This difference could be assigned neither to the charge nor to the size of the partitioned biomolecules since PAP and BR possess similar molecular weight (23,000) and isoelectric point (9.60). The presence of highly exposed tryptophans and positively charged residues (Lys, Arg and His) in BR molecule would be responsible for a charge transfer interaction between PEG and the protein and, therefore, the uneven distribution of BR in these systems.Fil: Rocha, María Victoria. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Nerli, Bibiana Beatriz. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin
Application of the aqueous two-phase systems of ethylene and propylene oxide copolymer-maltodextrin for protein purification
In this study, the effect of several factors that govern the partitioning behaviour of three model proteins, such as bovine serum albumin, lysozyme and trypsin was analysed in a two-phase system formed by maltodextrin and a copolymer of ethylene and propylene oxides. The protein partition coefficient (K r) showed to be very sensitive to temperature changes, protein molecular weight, pH medium and the lyotropic ion presence. The phase diagram obtained for these novel polymer-polymer two-phase systems shows two phases with high polymer concentrations. The maltodextrin is enriched in the bottom phase while the copolymer of ethylene and propylene oxides is found in the upper phase. Since this copolymer is thermoreactive, the upper phase can be removed and heated above the copolymer's cloud point resulting in the formation of a new two-phase system with a lower water phase, containing the target protein and an upper copolymer-rich phase. Our results show that systems formed by maltodextrin and a copolymer of ethylene and propylene oxides may be considered as an interesting alternative to be used in protein purification due to their low cost, and also because they offer a viable solution to problems of polymer removal and recycling.Fil: Bolognese, Belén. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; ArgentinaFil: Nerli, Bibiana Beatriz. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Picó, Guillermo Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentin