Liquid-liquid phase equilibria of aqueous biphasic systems based on glycerol formal:Application on tetracycline recovery from water

Biopharmaceuticals are commonly present in relatively low concentrations in aqueous solutions, making their detection and purification detrimental. In this work, we used novel aqueous biphasic systems based on glycerol formal (GF) to extract an important antibiotic - tetracycline. We report cloud points (solubility curve) and tie-lines for three ternary systems, containing GF, water, and inorganic salt (either K3PO4, K2HPO4, or K2CO3) at constant temperature of 298 K and at 0.1 MPa. The tie-line data of these ternary systems were correlated using the nonrandom two-liquid model, and binary interaction parameters of activity coefficients were estimated. The experimental and correlated tie-line data were compared in terms of average root-mean-square deviation and showed satisfactory agreements. The partition coefficients of tetracycline between two phases were measured, and corresponding extraction efficiencies were calculated. The maximum value of partition coefficient was 1551 for the system containing K3PO4, followed by values of 1145 and 927 for systems containing K2CO3 and K2HPO4, respectively. The calculated extraction efficiencies were very high - greater than 98.8%, demonstrating high potential for using aqueous biphasic systems based on GF for separation and purification processes

The effect of salts on the liquid–liquid phase equilibria of PEG600 + salt aqueous two-phase systems

Six new ATPSs were prepared by combining polyethylene glycol PEG600 with potassium citrate, dipotassium hydrogen phosphate, sodium formate, potassium formate, sodium sulfate, and lithium sulfate. Complete phase diagrams, including the binodal curve and three tie-lines, were determined at 23 °C. The experimental data obtained for the binodal curve were successfully adjusted to the Merchuk equation, and the reliability of tie-line data was confirmed using the equations suggested by Othmer–Tobias and Bancroft. The ability of each ion to induce ATPS formation was investigated. Na+ proved to be more effective in ATPS formation than K+ and Li+. For potassium salts, the order observed for the effectiveness of the anions was: HPO42– > C6H5O73– > HCO2–. Regarding the sodium salts, it was found that SO42– is clearly more effective than HCO2–. The position of the ions in the Hofmeister series and their free energy of hydration (ΔGhyd) were used to explain the ability of the ions to induce PEG salting-out. Furthermore, the effective excluded volume (EEV) of the salts was determined and the following order was found: Na2SO4 > K2HPO4 > Li2SO4 > K3C6H5O7 > NaCHO2 > KCHO2. Similar order was obtained when analyzing the size of the heterogeneous regions, suggesting the practical use of EEV as a comparison parameter between different ATPSs.This work is partially supported by project PEst-C/EQB/LA0020/2011, financed by FEDER through COMPETE-Programa Operacional Factores de Competitividade and by FCT-Fundacao para a Ciencia e a Tecnologia. Sara Silverio acknowledges her Ph.D. grant from FCT (SFRH/BD/43439/2008)

OptFlux: an open-source software platform for in silico metabolic engineering

<p>Abstract</p> <p>Background</p> <p>Over the last few years a number of methods have been proposed for the phenotype simulation of microorganisms under different environmental and genetic conditions. These have been used as the basis to support the discovery of successful genetic modifications of the microbial metabolism to address industrial goals. However, the use of these methods has been restricted to bioinformaticians or other expert researchers. The main aim of this work is, therefore, to provide a user-friendly computational tool for Metabolic Engineering applications.</p> <p>Results</p> <p><it>OptFlux </it>is an open-source and modular software aimed at being the reference computational application in the field. It is the first tool to incorporate strain optimization tasks, i.e., the identification of Metabolic Engineering targets, using Evolutionary Algorithms/Simulated Annealing metaheuristics or the previously proposed OptKnock algorithm. It also allows the use of stoichiometric metabolic models for (i) phenotype simulation of both wild-type and mutant organisms, using the methods of Flux Balance Analysis, Minimization of Metabolic Adjustment or Regulatory on/off Minimization of Metabolic flux changes, (ii) Metabolic Flux Analysis, computing the admissible flux space given a set of measured fluxes, and (iii) pathway analysis through the calculation of Elementary Flux Modes.</p> <p><it>OptFlux </it>also contemplates several methods for model simplification and other pre-processing operations aimed at reducing the search space for optimization algorithms.</p> <p>The software supports importing/exporting to several flat file formats and it is compatible with the SBML standard. <it>OptFlux </it>has a visualization module that allows the analysis of the model structure that is compatible with the layout information of <it>Cell Designer</it>, allowing the superimposition of simulation results with the model graph.</p> <p>Conclusions</p> <p>The <it>OptFlux </it>software is freely available, together with documentation and other resources, thus bridging the gap from research in strain optimization algorithms and the final users. It is a valuable platform for researchers in the field that have available a number of useful tools. Its open-source nature invites contributions by all those interested in making their methods available for the community.</p> <p>Given its plug-in based architecture it can be extended with new functionalities. Currently, several plug-ins are being developed, including network topology analysis tools and the integration with Boolean network based regulatory models.</p

Conversion of rice straw to bio-based chemicals: an integrated process using Lactobacillus brevis

Commercialization of lignocellulosic biomass as a feedstock for bio-based chemical production is problematic due to the high processing costs of pretreatment and saccharifying enzymes combined with low product yields. Such low product yield can be attributed, in large part, to the incomplete utilization of the various carbohydrate sugars found in the lignocellulosic biomass. In this study, we demonstrate that Lactobacillus brevis is able to simultaneously metabolize all fermentable carbohydrates in acid pre-processed rice straw hydrolysate, thereby allowing complete utilization of all released sugars. Inhibitors present in rice straw hydrolysate did not affect lactic acid production. Moreover, the activity of exogenously added cellulases was not reduced in the presence of growing cultures of L. brevis. These factors enabled the use of L. brevis in a process termed simultaneous saccharification and mixed sugar fermentation (SSMSF). In SSMSF with L. brevis, sugars present in rice straw hydrolysate were completely utilized while the cellulase maintained its maximum activity due to the lack of feedback inhibition from glucose and/or cellobiose. By comparison to a sequential hydrolysis and fermentation process, SSMSF reduced operation time and the amount of cellulase enzyme necessary to produce the same amount of lactic acid