Effect of process parameters and surfactant additives on the obtained activity of recombinant tryptophan hydroxylase (TPH1) for enzymatic synthesis of 5-hydroxytryptophan (5-HTP).

5-hydroxytryptophan (5-HTP) is an intermediate molecule in the biosynthesis of serotonin, an important neurotransmitter, regulating a series of metabolic and psychological functions in humans. In this work, we studied the heterologous production of Human tryptophan hydroxylase (TPH1) in Escherichia coli, for the synthesis of 5-hydroxytryptophan (5-HTP) from Tryptophan (Trp). To quantify TPH1 activity, a simple fluorescence-based microtiter plate assay was established, based on the changes in fluorescence emission at 340 nm between substrate and product when excited at 310 nm, allowing quick and reliable quantification of released 5-HTP.

Setting the Foundations of Aqueous Three-Phase Systems (A3PS) in the Quest for a Rational Design.

This work presents the first in-depth study of Aqueous Three-Phase Systems (A3PS) with the main purpose of unveiling their behaviour, hence contributing to the development of this new field. Thus, a complete definition of a quaternary system was carried through by describing all the regions in detail to represent them later on in a regular-tetrahedral diagram. The three aqueous faces of the tetrahedron demonstrated an undeviating influence in the segregation capacity.

Biorefining brewery spent grain polysaccharides through biotuning of ionic liquids.

Brewery spent grain (BSG), a relevant waste from beer industry mainly composed of polysaccharides and lignin, is experiencing a surge in the production with its associated environmental impact. Thus, this manuscript bets in the application of aqueous solutions of a cholinium-based ionic liquid (IL) containing glycinate as anion ([N][Gly]) for an efficient delignification pretreatment. The operation at 90 °C yielded drastic lignin reduction (75.

Molecular dynamics studies on the structure and interactions of ionic liquids containing amino-acid anions.

Several molecular dynamics (MD) simulations have been performed in order to obtain structural information on ionic liquids (ILs) based on amino-acid anions. Six hydrophilic ILs containing cholinium or imidazolium cations combined with alaninate, glycinate or lysinate anions were modelled using the all-atom CL&P and OPLS-AA force fields. Both pure ILs and their aqueous solutions have been studied.

Demonstrating the viability of halolipase production at a mechanically stirred tank biological reactor.

The definition of halophiles as "the coming stars of industrial biotechnology" in a recent review demands new research efforts for their efficient production at bioreactor scale. In this sense, the scarcity of information about halolipases production has furthered the research on the viability of Halomonas sp. LM1C culture in a mechanically stirred bioreactor.

Unravelling the suitability of biological induction for halophilic lipase production by Halomonas sp. LM1C cultures.

In this study, the viability of using biological induction as an alternative to the conventional chemical induction in lipase production by a novel halophilic microorganism, Halomonas sp. LM1C, has been demonstrated. Thus, a 9-times increase of lipase activity (3000U/L) was recorded when Staphylococcus equorum sp.

Scaling-up and ionic liquid-based extraction of pectinases from Aspergillus flavipes cultures.

The viability of the scaling-up of pectinases production by Aspergillus flavipes at 5L-bioreactor scale has been demonstrated by keeping constant the power input, and a drastic increase in the endo- and exopectinolytic enzyme production was recorded (7- and 40-fold, respectively). The main process variables were modelled by means of logistic and Gompertz equations. In order to overcome the limitations of the conventional downstream strategies, a novel extraction strategy was proposed on the basis of the adequate salting-out potential of two biocompatible cholinium-based ionic liquids (NCl and NHPO) in aqueous solutions of Tergitol, reaching more than 90% of extraction.

Simultaneous biotreatment of Polycyclic Aromatic Hydrocarbons and dyes in a one-step bioreaction by an acclimated Pseudomonas strain.

A Pseudomonas stutzeri strain acclimated to the presence of neoteric contaminants has been proposed for simultaneously remediating an effluent polluted with Polycyclic Aromatic Hydrocarbons and a diazo dye. The pollutants chemical nature imposed a strict control of both the medium composition and the operating conditions. pH, temperature and agitation rates of 7.

Pesticide removal from aqueous solutions by adding salting out agents.

Phase segregation in aqueous biphasic systems (ABS) composed of four hydrophilic ionic liquids (ILs): 1-butyl-3-methylimidazolium methylsulfate and 1-ethyl-3-methylimidazolium methylsulfate (C(n)C₁im C₁SO₄, n = 2 and 4), tributylmethyl phosphonium methylsulfate (P₄₄₄₁ C₁SO₄) and methylpyridinium methylsulfate (C₁Py C₁SO₄) and two high charge density potassium inorganic salts (K₂CO₃ and K₂HPO₄) were determined by the cloud point method at 298.15 K. The influence of the addition of the selected inorganic salts to aqueous mixtures of ILs was discussed in the light of the Hofmeister series and in terms of molar Gibbs free energy of hydration.

North Western Spain hot springs are a source of lipolytic enzyme-producing thermophilic microorganisms.

Several hot springs in Galicia (North Western Spain) have been investigated as potential sources of lipolytic enzyme-producing thermophilic microorganisms. After isolating 12 esterase producing strains, 9 of them were assured to be true lipase producers, and consequently grown in submerged cultures, obtaining high extracellular activities by two of them. Furthermore, a preliminary partial characterization of the crude lipase, obtained by ultrafiltration of the cell-free culture supernatant, was carried out at several pH and temperature values.

Scaling-up the production of thermostable lipolytic enzymes from Thermus aquaticus YT1.

The lipolytic enzymes synthesized by Thermusaquaticus YT1 present extremely interesting properties of thermostability (more than 70% of activity after 12 days at 80°C and a half-life time of 1 h at 95°C), which point out the interest of proposing efficient strategies to successfully tackle the scale-up of the production process. In this study,viable scaling-up of the production process was implemented,and relevant aspects affecting the enzyme synthesis, such as the mineral composition of the culture medium, the aeration and the agitation have been evaluated.A strategy combining the modification of the culture medium and the aeration degree was also approached by adding perfluorocarbons, compounds which improve the availability of oxygen in the culture medium.

A process for extracellular thermostable lipase production by a novel Bacillus thermoamylovorans strain.

A lipolytic enzyme-producing thermophilic microorganism, recently isolated from a hot spring in Galicia (North Western Spain), has been investigated. First, the strain was genetically identified and tentatively named Bacillus thermoamylovorans CH6B. It produced significant levels (around 450 U/L) of extracellular lipolytic activity in shake flask cultures, and the most suitable conditions for this biological process were found at temperatures between 50 and 55 °C, and an initial pH value around 7.

New insight into phase equilibria involving imidazolium bistriflamide ionic liquids and their mixtures with alcohols and water.

The fluid phase equilibria (liquid-liquid demixing behavior (LLE)) of mixtures of ionic liquids of the 1-alkyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide family, [C(n)mim][NTf(2)], with 2-methylpropanol or n-octanol were investigated. Binary mixtures of [C(4)mim][NTf(2)] + alcohol and [C(6)mim][NTf(2)] + alcohol were compared to pseudobinary mixtures of (0.5[C(2)mim] + 0.

Phase equilibria of haloalkanes dissolved in ethylsulfate- or ethylsulfonate-based ionic liquids.

The temperature-composition phase diagrams of 40 binary mixtures composed of a haloalkane dissolved in either 1-ethyl-3-methylimidazolium ethylsulfate or 1-ethyl-3-methylimidazolium ethylsulfonate were measured from ambient temperature to the boiling point temperature of the solute. The coexistence curves corresponding to liquid-liquid equilibria (LLE) boundaries were visually determined and the experimental results have been correlated using either the nonrandom two-liquid (NRTL) model or a set of empirical equations capable of describing the corresponding upper critical solution temperatures (UCSTs) loci. The different types of LLE behavior were discussed in terms of the type of ionic liquid solvent, the alkyl-chain length of the solute, and the type and pattern of halogen substitution present in the haloalkane.

Strategies for utilisation of food-processing wastes to produce lipases in solid-state cultures of Rhizopus oryzae.

The aim of the present study was to investigate the feasibility of several food-processing wastes as support substrate for lipolytic enzymes production by the fungus Rhizopus oryzae under solid-state conditions. Different experiments were conducted to select the variables that allow obtaining high levels of lipolytic enzyme activity. In particular, the use of inert and non-inert solid materials and lipidic and surfactant compounds was evaluated.

Thermostable lipolytic enzymes production in batch and continuous cultures of Thermus thermophilus HB27.

Several studies in laboratory-scale bioreactors are undertaken in order to verify the beneficial effect of thermal spring water in the culture medium of Thermus thermophilus HB27. Two bioreactor configurations, stirred tank and airlift, are investigated to determine the most suitable one to develop a continuous process. Water mineral composition affects the lipolytic enzyme secretion and growth of T.

Strategies for improving extracellular lipolytic enzyme production by Thermus thermophilus HB27.

In Thermus thermophilus HB27 cultures the localisation of lipolytic activity is extracellular, intracellular and membrane bound, with low percentage for the former. Therefore, the extracellular secretion must be increased in order to simplify the downstream process and to reduce the economic cost. This study focuses on the design of an innovative operational strategy to increase extracellular lipolytic enzyme production by T.