Carlota de Oliveira Rangel-Yagui

Effect of reducing the light intensity on the growth and production of chlorophyll by Spirulina platensis

Abstract The influence of light intensity reduction on Spirulina platensis cultivation was investigated, using urea and KNO 3 as nitrogen sources. The reduction of light intensity from 5 to 2 klux was studied both on the 9th and the 13th day of cultivation. Increases of up to 29% in the total chlorophyll production were observed for the cultivations with light intensity reduction, in comparison with the cultivations carried out at fixed light intensities.

Micellar solubilization of ibuprofen: influence of surfactant head groups on the extent of solubilization

An important property of micelles with particular significance in pharmacy is their ability to increase the solubility of poorly soluble drugs in water, thus increasing their bioavailability. In this work, the solubilization of ibuprofen (IBU) was studied in micellar solutions of three surfactants possessing the same hydrocarbon tail but different hydrophilic head groups, namely sodium dodecyl sulphate (SDS), dodecyltrimethylammonium bromide (DTAB), and n-dodecyl octa(ethylene oxide) (C12EO8). The results showed that, irrespective of the surfactant type, the solubility of IBU increased linearly with increasing surfactant concentration, as a consequence of the association between the drug and the micelles. The 80 mM DTAB and the 80 mM C12EO8 micellar solutions resulted in a 16-fold increase in solubility of IBU when compared to the buffer solution, whereas the 80 mM SDS micellar solution resulted in a 5.5-fold increase in IBU solubility. The highest value of molar solubilization capacity (c) was obtained with DTAB, c = 0.97, followed by C12EO8 ,c = 0.72, and finally SDS, c = 0.23. However, due to the stronger tendency of the nonionic surfactant in forming micelles in solution, at the same surfactant concentration, we obtained the same solubility of IBU in both DTAB and C12EO8.

LPS removal from an E. coli fermentation broth using aqueous two-phase micellar system

In biotechnology, endotoxin (LPS) removal from recombinant proteins is a critical and challenging step in the preparation of injectable therapeutics, as endotoxin is a natural component of bacterial expression systems widely used to manufacture therapeutic proteins. The viability of large‐scale industrial production of recombinant biomolecules of pharmaceutical interest significantly depends on the separation and purification techniques used. The aim of this work was to evaluate the use of aqueous two‐phase micellar system (ATPMS) for endotoxin removal from preparations containing recombinant proteins of pharmaceutical interest, such as green fluorescent protein (GFPuv). Partition assays were carried out initially using pure LPS, and afterwards in the presence of E. coli cell lysate. The ATPMS technology proved to be effective in GFPuv recovery, preferentially into the micelle‐poor phase (KGFPuv < 1.00), and LPS removal into the micelle‐rich phase (%REMLPS > 98.00%). Therefore, this system can be exploited as the first step for purification in biotechnology processes for removal of higher LPS concentrations.

Cruzain inhibition by hydroxymethylnitrofurazone and nitrofurazone: investigation of a new target in Trypanosoma cruzi

Nitrofurazone (NF) and its derivative, hydroxymethylnitrofurazone (NFOH), have presented antichagasic activity. NFOH has higher activity and lower mutagenicity. The aim of this work was to assess whether NF and its derivative NFOH would also be inhibitors of cruzain, besides their trypanothione reductase inhibitory activity. In vitro cruzain inhibition tests were performed for both compounds, and the 50% inhibitory concentration (IC50) for NF and NFOH presented values of 22.83 ± 1.2 μM and 10.55 ± 0.81 μM, respectively. AM1 semi-empirical molecular modeling studies were performed to understand the activity of the compounds, corroborating the observed cruzain inhibitory activity.

Therapeutic l-asparaginase: upstream, downstream and beyond

Abstract l-asparaginase (l-asparagine amino hydrolase, E.C.3.5.1.1) is an enzyme clinically accepted as an antitumor agent to treat acute lymphoblastic leukemia and lymphosarcoma. It catalyzes l-asparagine (Asn) hydrolysis to l-aspartate and ammonia, and Asn effective depletion results in cytotoxicity to leukemic cells. Microbial l-asparaginase (ASNase) production has attracted considerable attention owing to its cost effectiveness and eco-friendliness. The focus of this review is to provide a thorough review on microbial ASNase production, with special emphasis to microbial producers, conditions of enzyme production, protein engineering, downstream processes, biochemical characteristics, enzyme stability, bioavailability, toxicity and allergy potential. Some issues are also highlighted that will have to be addressed to achieve better therapeutic results and less side effects of ASNase use in cancer treatment: (a) search for new sources of this enzyme to increase its availability as a drug; (b) production of new ASNases with improved pharmacodynamics, pharmacokinetics and toxicological profiles, and (c) improvement of ASNase production by recombinant microorganisms. In this regard, rational protein engineering, directed mutagenesis, metabolic flux analysis and optimization of purification protocols are expected to play a paramount role in the near future.

Growth and content of Spirulina platensis biomass chlorophyll cultivated at different values of light intensity and temperature using different nitrogen sources

The effects of light intensity and temperature in S. platensis cultivation with potassium nitrate or urea as nitrogen source were investigated, as well as the biomass chlorophyll contents of this cyanobacteria, through the Response Surface Methodology. Experiments were performed at temperatures from 25 to 34.5ºC and light intensities from 15 to 69 µmol photons m−2 s−1, in mineral medium. In cultivations with both sources of nitrogen, KNO3 and urea, statistic evaluation through multiple regression, no interactions of such independent variables were detected in the results of the dependent variables maximum cell concentration, chlorophyll biomass contents, cell and chlorophyll productivities, as well as in the nitrogen-cell conversion factor. In cultivation performed with both sources of nitrogen, it was possible to obtain satisfactory adjustments to relate the dependent variables to the independent variables. The best results were achieved at temperature of 30ºC, at light intensity of 60 µmol photons m−2s−1, for cell growth, with cell productivity of approximately 95 mg L−1 d−1 in cultivations with urea. For the chlorophyll biomass content, the most adequate light intensity was 24 µmol photons m−2 s−1.

Can affinity interactions influence the partitioning of glucose-6-phosphate dehydrogenase in two-phase aqueous micellar systems?

In this work, we provide an investigation of the role and strength of affinity interactions on the partitioning of the glucose-6-phosphate dehydrogenase in aqueous two-phase micellar systems. These systems are constituted of micellar surfactant solutions and offer both hydrophobic and hydrophilic environments, providing selectivity to biomolecules. We studied G6PD partitioning in systems composed of the nonionic surfactants, separately, in the presence and absence of affinity ligands. We observed that G6PD partitions to the micelle-poor phase, owing to the strength of excluded-volume interactions in these systems that drive the protein to the micelle-poor phase, where there is more free volume available.

Polymeric micelles and molecular modeling applied to the development of radiopharmaceuticals

Micelles composed of amphiphilic copolymers linked to a radioactive element are used in nuclear medicine predominantly as a diagnostic application. A relevant advantage of polymeric micelles in aqueous solution is their resulting particle size, which can vary from 10 to 100 nm in diameter. In this review, polymeric micelles labeled with radioisotopes including technetium (99mTc) and indium (111In), and their clinical applications for several diagnostic techniques, such as single photon emission computed tomography (SPECT), gamma-scintigraphy, and nuclear magnetic resonance (NMR), were discussed. Also, micelle use primarily for the diagnosis of lymphatic ducts and sentinel lymph nodes received special attention. Notably, the employment of these diagnostic techniques can be considered a significant tool for functionally exploring body systems as well as investigating molecular pathways involved in the disease process. The use of molecular modeling methodologies and computer-aided drug design strategies can also yield valuable information for the rational design and development of novel radiopharmaceuticals.

Biopharmaceuticals from microorganisms: from production to purification

The use of biopharmaceuticals dates from the 19th century and within 5–10 years, up to 50% of all drugs in development will be biopharmaceuticals. In the 1980s, the biopharmaceutical industry experienced a significant growth in the production and approval of recombinant proteins such as interferons (IFN α, β, and γ) and growth hormones. The production of biopharmaceuticals, known as bioprocess, involves a wide range of techniques. In this review, we discuss the technology involved in the bioprocess and describe the available strategies and main advances in microbial fermentation and purification process to obtain biopharmaceuticals.

A 2D-QSPR approach to predict blood-brain barrier penetration of drugs acting on the central nervous system

Farmacos que atuam no sistema nervoso central (SNC) devem atravessar a barreira hematoencefalica (BHE) para exercerem suas acoes farmacologicas. A difusao passiva atraves da BHE pode ser parcialmente expressa pelo coeficiente de particao entre os compartimentos encefalico e sanguineo (logBB, brain/blood partition coefficient). Considerando-se que a avaliacao experimental de logBB e dispendiosa e demorada, metodos teoricos como estudos das relacoes entre estrutura quimica e propriedade (QSPR, Quantitative Structure-Property Relationships) podem ser utilizados na previsao dos valores de logBB. Neste estudo, uma abordagem de QSPR-2D foi aplicada a um conjunto de 28 moleculas com acao central, usando logBB como propriedade biologica. O melhor modelo de QSPR [n = 21, r = 0,94 (r² = 0,88), s = 0,28 e Q² = 0,82] apresentou tres descritores moleculares: o coeficiente calculado de particao n-octanol/agua (ClogP), area de superficie polar (PSA) e polarizabilidade (α). Seis dos sete compostos do conjunto de avaliacao foram bem previstos pelo modelo, o que corresponde a um bom poder de previsao externa (85,7%). Os resultados obtidos podem auxiliar de forma relevante em estudos futuros, orientando quais descritores moleculares devem ser considerados para estimar logBB e prever a passagem atraves da BHE de moleculas estruturalmente relacionadas as do conjunto investigado.