Expression and characterization of an N-truncated form of the NifA protein of Azospirillum brasilense

Azospirillum brasilense is a nitrogen-fixing bacterium associated with important agricultural crops such as rice, wheat and maize. The expression of genes responsible for nitrogen fixation (nif genes) in this bacterium is dependent on the transcriptional activator NifA. This protein contains three structural domains: the N-terminal domain is responsible for the negative control by fixed nitrogen; the central domain interacts with the RNA polymerase σ54 co-factor and the C-terminal domain is involved in DNA binding. The central and C-terminal domains are linked by the interdomain linker (IDL). A conserved four-cysteine motif encompassing the end of the central domain and the IDL is probably involved in the oxygen-sensitivity of NifA. In the present study, we have expressed, purified and characterized an N-truncated form of A. brasilense NifA. The protein expression was carried out in Escherichia coli and the N-truncated NifA protein was purified by chromatography using an affinity metal-chelating resin followed by a heparin-bound resin. Protein homogeneity was determined by densitometric analysis. The N-truncated protein activated in vivo nifH::lacZ transcription regardless of fixed nitrogen concentration (absence or presence of 20 mM NH4Cl) but only under low oxygen levels. On the other hand, the aerobically purified N-truncated NifA protein bound to the nifB promoter, as demonstrated by an electrophoretic mobility shift assay, implying that DNA-binding activity is not strictly controlled by oxygen levels. Our data show that, while the N-truncated NifA is inactive in vivo under aerobic conditions, it still retains DNA-binding activity, suggesting that the oxidized form of NifA bound to DNA is not competent to activate transcription.

Avaliação da influência dos milhos QPM nas características sensoriais de bolo

Atualmente já não existem mais dúvidas sobre o potencial nutricional dos milhos QPM na alimentação humana. No entanto pouco se conhece sobre os milhos QPM (Quality Protein Maize) na produção de alimentos. O presente trabalho teve como objetivo avaliar o efeito da adição de diferentes variedades de milho nas características de aparência, aroma, sabor e textura de bolo de fubá utilizando análise descritiva quantitativa. Sete formulações foram avaliadas por uma equipe de provadores treinados, onze atributos sensoriais foram definidos. Após um período de treinamento, os provadores avaliaram as amostras através de delineamento de blocos incompletos balanceados usando escala não estruturada de 9cm. As mesmas foram também avaliadas em relação a aceitabilidade através da escala hedônica de 9 pontos, por 70 consumidores de bolo de fubá. Os resultados foram analisados através da ANOVA seguida pelo teste de Tukey. As amostras apresentaram diferença significativa (p<0,05) em todos os atributos avaliados. As formulações diferiram quanto à aceitabilidade (p<0,05). Estas se situaram entre os termos hedônicos "gostei ligeiramente" e "gostei moderadamente"

Amido resistente, a última geração no controle de energia e digestão saudável

Estudos clínicos demonstraram que amido resistente tem propriedades semelhantes a fibras e mostra benefícios fisiológicos em humanos, podendo resultar em prevenção de doenças. A Organização Mundial de Saúde (OMS) recomenda que cerca de 55% da energia ingerida seja proveniente de carboidratos. A preocupação com o tipo de carboidrato ingerido é importante, pois há fibras, por exemplo, que devem ser ingeridas de 25-30 g/dia/pessoa, e normalmente não o são. Por ser um alimento resistente à digestão e fermentado no intestino grosso, principalmente pelas bifidobactérias, o amido resistente é um alimento prebiótico. Durante a fermentação ocorre a produção de ácidos graxos de cadeia curta, principalmente o butirato, que contribui muito para a saúde do cólon, inibindo o crescimento de células cancerígenas devido à redução do pH no intestino grosso. Além disso, contribui para a produção da energia difusa progressiva (EDP), que é a energia liberada ao longo do tempo de uma digestão lenta, e para a queda do índice glicêmico dos alimentos, proporcionando uma menor resposta glicêmica e, conseqüentemente, uma resposta insulínica mais adequada, auxiliando no tratamento da diabete, principalmente do tipo 2 e mantendo o indivíduo com sensação de saciedade por um período maior de tempo. A National Starch Food Innovation possui o Hi-Maize 260 como amido resistente que, quando comparado com as fibras convencionais, apresenta muitas vantagens. É branco e possui sabor brando, tamanho pequeno de partícula e baixa capacidade de retenção de água, praticamente não altera a textura de produtos de baixa umidade como pães, macarrão, barrinha de cereais, etc, ao contrário das fibras convencionais, o que permite formular produtos com alto teor de fibras e ainda ser rotulado como simplesmente "amido de milho". Também apresenta teor calórico reduzido e pode ser usado como agente de corpo complementar em formulações com valor reduzido ou sem gordura.

Obtention and characterization of gluten-free baked products

This work had as objective the development of gluten-free breads and muffins using rice flour and maize and cassava starches. From seven samples resulting from a Simplex-Centroid design, the sensory and instrumental analyses of specific volume, elasticity, and firmness were performed. For the sensory analysis, the optimized formulation contained 50% of rice flour and 50% of cassava starch, and for the instrumental evaluation, the optimal simultaneous point for the three conducted analyses were 20% of rice flour, 30% of cassava starch, and 50% of maize starch. A comparative analysis of specific volume, elasticity, firmness, and triangular test was performed with pre-baked, baked, and frozen bread. Physicochemical, nutritional, and microbiological analyses were performed for both bread and muffin according to the Brazilian legislation.

Corn germ with pericarp in relation to whole corn: nutrient contents, food and protein efficiency, and protein digestibility-corrected amino acid score

The germ fraction with pericarp (bran) is generated in the industrial processing of corn kernel, and it is used for oil extraction and animal feed. This study evaluated the nutritional and protein quality of this fraction in relation to whole corn. The proximate composition, mineral contents, and amino acid profile of the germ fraction with pericarp and of whole corn were determined. A 4-week experiment was conducted using 36 weanling male Wistar rats, and three 10%-protein diets (reference, germ with 15% lipids and casein with 15% lipids), two 6%-protein diets (whole corn and casein), and a protein-free diet were prepared. The germ showed higher contents of proteins, lipids, dietary fiber (27.8 g.100 g-1), ash, minerals (Fe and Zn- approximately 5 mg.100 g-1), and lysine (57.2 mg.g-1 protein) than those of corn. The germ presented good quality protein (Relative Protein Efficiency Ratio-RPER = 80%; Protein Digestibility-Corrected Amino Acid Score-PDCAAS = 86%), higher than that of corn (RPER = 49%; PDCAAS = 60%). The corn germ fraction with pericarp is rich in dietary fiber, and it is a source of good quality protein as well as of iron and zinc, and its use as nutritive raw material is indicated in food products for human consumption.

Sugarcane starch: quantitative determination and characterization

Starch is found in sugarcane as a storage polysaccharide. Starch concentrations vary widely depending on the country, variety, developmental stage, and growth conditions. The purpose of this study was to determine the starch content in different varieties of sugarcane, between May and November 2007, and some characteristics of sugarcane starch such as structure and granules size; gelatinization temperature; starch solution filterability; and susceptibility to glucoamylase, pullulanase, and commercial bacterial and fungal α-amylase enzymes. Susceptibility to debranching amylolytic isoamylase enzyme from Flavobacterium sp. was also tested. Sugarcane starch had spherical shape with a diameter of 1-3 µm. Sugarcane starch formed complexes with iodine, which showed greater absorption in the range of 540 to 620 nm. Sugarcane starch showed higher susceptibility to glucoamylase compared to that of waxy maize, cassava, and potato starch. Sugarcane starch also showed susceptibility to debranching amylolytic pullulanases similar to that of waxy rice starch. It also showed susceptibility to α-amylase from Bacillus subtilis, Bacillus licheniformis, and Aspergillus oryzae similar to that of the other tested starches producing glucose, maltose, maltotriose, maltotetraose, maltopentose and limit α- dextrin.

Optimization of extrusion process for production of nutritious pellets

A blend of 50% Potato Starch (PS), 35% Quality Protein Maize (QPM), and 15% Soybean Meal (SM) were used in the preparation of expanded pellets utilizing a laboratory extruder with a 1.5 × 20.0 × 100.0 mm die-nozzle. The independent variables analyzed were Barrel Temperature (BT) (75-140 °C) and Feed Moisture (FM) (16-30%). The effect of extrusion variables was investigated in terms of Expansion Index (EI), apparent density (ApD), Penetration Force (PF) and Specific Mechanical Energy (SME), viscosity profiles, DSC, crystallinity by X-ray diffraction, and Scanning Electronic Microscopy (SEM). The PF decreased from 30 to 4 kgf with the increase of both independent variables (BT and FM). SME was affected only by FM, and decreased with the increase in this variable. The optimal region showed that the maximum EI was found for BT in the range of 123-140 °C and 27-31% for FM, respectively. The extruded pellets obtained from the optimal processing region were probably not completely degraded, as shown in the structural characterization. Acceptable expanded pellets could be produced using a blend of PS, QPM, and SM by extrusion cooking.