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

Retrospective evaluation of whole exome and genome mutation calls in 746 cancer samples

Funder: NCI U24CA211006Abstract: The Cancer Genome Atlas (TCGA) and International Cancer Genome Consortium (ICGC) curated consensus somatic mutation calls using whole exome sequencing (WES) and whole genome sequencing (WGS), respectively. Here, as part of the ICGC/TCGA Pan-Cancer Analysis of Whole Genomes (PCAWG) Consortium, which aggregated whole genome sequencing data from 2,658 cancers across 38 tumour types, we compare WES and WGS side-by-side from 746 TCGA samples, finding that ~80% of mutations overlap in covered exonic regions. We estimate that low variant allele fraction (VAF < 15%) and clonal heterogeneity contribute up to 68% of private WGS mutations and 71% of private WES mutations. We observe that ~30% of private WGS mutations trace to mutations identified by a single variant caller in WES consensus efforts. WGS captures both ~50% more variation in exonic regions and un-observed mutations in loci with variable GC-content. Together, our analysis highlights technological divergences between two reproducible somatic variant detection efforts

Screening of different agroindustrial by-products for industrial enzymes production by fermentation processes

Agroindustrial by-products are an abundant source of biocompounds that contain valuable nutrients, which are not exploited. In this work, lignocellulosic wastes (LW) were used in submerged fermentation (SmF) and solid-state fermentation (SSF) by Aspergillus niger NRRL3 to obtain valuable enzymes required in industries. SmF using soya bean hulls (SH), wheat bran (WB) and a by-product of wheat flour (F) produced the highest activities of endo-1,4-β-xylanase (Xyl) and endo-1,4-β-D-glucanase (EG) being at least 3 times lower than those obtained by SSF. The highest ratio of Xyl to EG was obtained in SmF with F. Xyl obtained by SmF with WB was the most thermally resistant. The enzymatic extract obtained in SmF using SH presented a high power of saccharification. The production of enzymes for further application such as bioethanol generation process revalue these LW and can help offset growing environmental problems.Fil: Taddia, Antonela. 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: Boggione, María Julia. 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: Tubio, Gisela. 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

Structural and functional analysis of Aspergillus niger xylanase to be employed in polyethylenglycol/salt aqueous two-phase extraction

The structure and enzymatic activity of Aspergillus niger xylanase were evaluated in different media to establish an appropriate protocol for the extraction of the enzyme in polymer/salt aqueous two-phase systems. Different factors were studied: the concentration and molecular weight (1000, 2000, 4600 and 8000) of polyethyleneglycol, the concentration and type of salt (sodium citrate and potassium phosphate) and pH, time and temperature. Xylanase was stable for 5 h at pH between 2.7 and 9.0 and at temperatures up to 50 °C. Fluorescence spectroscopy and circular dichroism experiments showed that neither the secondary/tertiary structure of the enzyme nor its catalytic activity were significantly altered in the presence of either salt or PEG. Xylanase partitioned into the PEG-rich phase driven by the excluded volume effect. Partitioning was more favorable to the polymer phase in the PEG1000/NaCit system, where Kp was 12 times higher than in the others aqueous two-phase systems. These results demonstrate the potential application of the PEG1000/NaCit system as a first step for the extraction of Aspergillus niger xylanase.Fil: Loureiro, Dana Belen. 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: Romanini, Diana. 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: Tubio, Gisela. 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

Enzymatic cocktail production by a co-cultivation Solid-State Fermentation for detergent formulation

The industry of detergents is one of the main claimers of enzymes. Here we show a promising technique, a co-cultivation Solid-State Fermentation (SSF) of Aspergillus niger and Aspergillus oryzae for the production of enzymatic cocktails. It was optimized by a central composite design where a proportion 75 % of wheat bran and 25 % of soybean husk as substrate and a conidia ratio 25 % A. niger and 75 % A. oryzae were the optimal conditions. Furthermore, the cocktail was analyzed by Nanoscale liquid chromatography coupled to tandem mass spectrometry (Nano LC-MS/MS) and it was concluded that 54.17 % of proteins belong to the enzymes of interest, where 28.79 % were amylases (308.47 IU/mL), 10.56 % were proteases (2228 IU/mL,), 13.47 % were cellulases (3.97 IU/mL) and 1.34 % were lipases (0.789 IU/mL). Finally, the enzymes presented a great stability in a wide range of pH and temperatures; and an excellent cleaning efficiency against cleaning control strips indicating that they could be used for formulations.Fil: Morilla, Esteban Amador. 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: Mutti Stegmann, Paula. 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: Tubio, Gisela. 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

Relationship between the protein surface hydrophobicity and its partitioning behaviour in aqueous two-phase systems of polyethyleneglycol- dextran

In order to develop possible correlations to predict partioning behaviour of proteins, five mammalian albumins (goat, bovine, equine, human and pig ones) with similar physico-chemical properties (molecular mass and isoelectrical point) were chosen. Evaluation of the relationship between hydrophobicity and partitioning coefficient (Kr) in polyethylenglycol-dextran (PEG-DxT500) systems formed by polyethyleneglycols of different molecular mass (3350, 6000 and 10,000) was investigated by estimating relative surface hydrophobicity (So) with a fluorescent probe, 1 anilino-8-naphthalene sulfonate. No relationship between Kr and So was found for systems formed by PEG3350, while aqueous two-phase systems with PEG6000 and PEG10,000 gave better correlations. The results obtained may be explained on the basis of an increase in the interaction between the latter PEGs and the protein due to their higher hydrophobic character which increases as the PEG molecular mass does so. In this way, systems with PEGs of higher molecular mass give the highest resolution to exploit hydrophobicity in partitioning.Fil: Tubio, Gisela. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Departamento de Química y Física. Área Fisicoquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Nerli, Bibiana Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Departamento de Química y Física. Área Fisicoquímica; ArgentinaFil: Picó, Guillermo Alfredo. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Departamento de Química y Física. Área Fisicoquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentin

Insights on fungal solid-state fermentation for waste valorization: Conidia and chitinase production in different reactor configurations

Different reactor configurations are paired with a wide variety of agro-industrial wastes of different biodegradability to produce fungal conidia by solid-state fermentation. This work presents a preliminary comparative study between packed-bed and tray reactor configurations to produce Beauveria bassiana and Trichoderma harzianum conidia using two different substrates in terms of biodegradability: rice husk or beer draff complemented with wood chips. Conidia production, mean temperature and respiration indexes have been analysed in most of the presented reactor configurations. Both strains showed higher conidia production when using beer draff complemented with wood chips as substrate due to the use of a mixture as substrate. When working with beer draff, chitinase analyses obtained similar profiles in both strains but higher overall values using TH. Conidia and chitinase production maximums were not achieved at the same time, having 2–3 days of difference depending on the strain. No significant differences in mean temperature were shown between most of the performed fermentations. As a result of the present work, further scaling of both packed bed and tray configurations using beer draff and wood chips to produce BB or TH conidia would be advisable. More experiments should be performed to optimize both conidia and chitinase productions to enhance the quality of the final product.Fil: Sal, A. Universidad Autónoma de Barcelona. Departamento de Ingeniería Química, Biológica y Ambiental. Grupo de Investigación en Compostaje. España.Fil: Echegaray, T. Universidad Autónoma de Barcelona. Departamento de Ingeniería Química, Biológica y Ambiental. Grupo de Investigación en Compostaje. España.Fil: Boggione, María Julia. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos (IPROByQ-CONICET). Argentina.Fil: Tubio, Gisela. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos (IPROByQ-CONICET). Argentina.Fil: Barrena, R. Universidad Autónoma de Barcelona. Departamento de Ingeniería Química, Biológica y Ambiental. Grupo de Investigación en Compostaje. España.Fil: Artola, A. Universidad Autónoma de Barcelona. Departamento de Ingeniería Química, Biológica y Ambiental. Grupo de Investigación en Compostaje. España

Partitioning of xylanase from Thermomyces lanuginosus in PEG/NaCit aqueous two-phase systems: Structural and functional approach

The structure and catalytic activity of xylanase from Thermomyces lanuginosus were studied in different media (containing polyethylene glycol -PEG- or salt) at different temperatures. The aim was to study how the native structure of the enzyme is affected to understand the partitioning behavior of xylanase in PEG/sodium citrate (PEG/NaCit) aqueous two-phase systems. The presence of PEGs of different molar masses slightly altered the native structure of xylanase, although its catalytic activity was not affected. All the polymers assayed protect the native structure (and catalytic activity) of xylanase against temperature, except for PEG1000. Surface hydrophobicity experiments showed that xylanase favorable interacts with PEGs. Partitioning experiments confirmed this result and demonstrated that PEG1000/NaCit is the best system to partition xylanase from Thermomyces lanuginosus, since the Kp was 17.7 ± 0.3.Fil: Loureiro, Dana Belen. 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: Braia, Mauricio Javier. 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: Romanini, Diana. 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: Tubio, Gisela. 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

Mixed Cultures of Aspergillus niger and Rhizopus oryzae Using Lignocellulosic Substrates to Improve Hydrolytic Enzyme Production

This study aims to evaluate the potential of filamentous fungi consortiums to design a bioprocess for producing hydrolytic enzymatic cocktails on lignocellulosic biomass. Four different microorganisms were analyzed in solid and submerged monoculture fermentation systems: Aspergillus niger, Trichoderma harzianum, Rhizopus oryzae, and Thermomyces lanuginosus. A. niger and R. oryzae were selected as the most suitable to form a consortium. Two different inoculation strategies were assessed, one was simultaneous and the other was a 12-h post-phase inoculation. Xylanase, cellulase, amylase, pectinase, and protease activities were evaluated in mono and mixed cultures. Furthermore, the secretomes of the fermentation systems were analyzed by mass spectrometry to compare and determine the difference in the protein profiles excreted. The mixed culture of A. niger and R. oryzae, which were inoculated simultaneously in solid-state fermentation, showed the highest levels of enzyme at 96 h, 30°C, 1 × 107 conidia, and 65% relative humidity. The proportion of xylanase (98 IU/gds), cellulase (27 IU/gds), amylase (30 IU/gds), pectinase (21 IU/gds) and protease (108,000 IU/gds) was enhanced by 25% regarding the monoculture, demonstrating that the use of microbial consortiums can be a promising alternative to obtain enzymatic cocktails with high synergism.Fil: Morilla, Esteban Amador. 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: Taddia, Antonela. 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: Sortino, Maximiliano Andrés. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Tubio, Gisela. 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

Characterization of the Interaction Between Pancreatic Trypsin and an Enteric Copolymer as a Tool for Several Biotechnological Applications

Protein-polyelectrolyte complexes are very interesting systems since they can be applied in many long-established and emerging areas of biotechnology. From nanotechnology to industrial processing, these complexes are used for many purposes: to build multilayer particles for biosensors; to entrap and deliver proteins for pharmaceutical applications; to isolate and immobilize proteins. The enteric copolymer poly(methacrylic acid-co-methyl methacrylate) 1:2 (MMA) has been designed for drug delivery although its chemical properties allow to use it for other applications. Understanding the interaction between trypsin and this polymer is very important in order to optimize the mechanism of formation of this complex for different biotechnological applications.The formation of the trypsin-MMA complex was studied by spectroscopy and isothermal titration calorimetry. Structural analysis of trypsin was carried out by catalytic activity assays, circular dichroism and differential scanning calorimetry. Isothermal titration calorimetry experiments showed that the insoluble complex contains 12 trypsin molecules per MMA molecule at pH 5 and they interact with high affinity to form insoluble complexes. Both electrostatic and hydrophobic forces are involved in the formation of the complex. The structure of trypsin is not affected by the presence of MMA, although it interacts with some domains of trypsin affecting its thermal denaturation as seen in the differential scanning calorimetry experiments. Its catalytic activity is not altered. Dynamic light scattering demonstrated the presence of a soluble trypsin-copolymer complex at pH 5 and 8. Turbidimetric assays show that the insoluble complex can be dissolved by low ionic strength and/or pH in order to obtain free native trypsin.Fil: Braia, Mauricio Javier. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Rosario. Instituto de Procesos Biotecnologicos y Quimicos Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; ArgentinaFil: Loureiro, Dana Belen. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Rosario. Instituto de Procesos Biotecnologicos y Quimicos Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; ArgentinaFil: Tubio, Gisela. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Rosario. Instituto de Procesos Biotecnologicos y Quimicos Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; ArgentinaFil: Romanini, Diana. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Rosario. Instituto de Procesos Biotecnologicos y Quimicos Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentin