O agronegocio brasileiro possui grande competitividade internacional, principalmente pelos grandes esforcos historicamente dispendidos pelos institutos publicos de pesquisa e Universidades. No entanto, muito dos avancos obtidos pelo agronegocio brasileiro e decorrente do uso em larga escala de fertilizantes e agroquimicos, que por sua vez tem que ser importados em grandes quantidades para suprir a demanda dos agricultores brasileiros. Neste sentido, a seguranca e a sustentabilidade do agronegocio brasileiro esta ameacada pela susceptibilidade destes insumos aos precos do mercado internacional, alem de estar fundamentado na confianca de entrega destes insumos pelos fornecedores. Em adicao, a sociedade global apresenta uma nova apreensao quanto a necessidade de alcancar produtividades mais elevadas e de uma maneira mais sustentavel, para prover a populacao crescente com alimentos e energia. As relacoes naturais existentes entre as plantas e a microbiota do solo estao no cerne desta questao, uma vez que a quantidade de informacoes cientificas disponibilizadas pela literatura mundial dando suporte ao potencial real de aplicacao destas associacoes em beneficio da producao de alimentos e energia e crescente. A substituicao total do uso de fertilizantes nitrogenados pela fixacao biologica de nitrogenio realizada pelos rizobios, nos cultivos de soja realizados no Brasil, corrobora a eficiencia destas associacoes e imprimem a necessidade de expandir os mecanismos relacionados com o suprimento de nutrientes para diferentes culturas nao-leguminosas. Para o alcance destes objetivos, sao necessarios ainda muitos esforcos de pesquisa para um melhor entendimento dos mecanismos que controlam estas interacoes naturais entre plantas e a microbiota associativa, como tambem para o desenvolvimento de formulacoes de biofertilizantes que apresentem uma elevada eficiencia sob uma grande variedade de condicoes edafoclimaticas, como as que ocorrem no Brasil. Algumas abordagens biotecnologicas dirigidas para o alcance destes objetivos sao revisadas e discutidas ao longo desta revisao.
This study aimed to compare several DNA polymerases in order to optimize costs of microsatellite amplification reaction through the PowerPlex®16 BIO System. Three different polymerase brands have been used: (i) Paq5000 DNA Polymerase (Stratagene) and (ii) Platinum® Taq DNA Polymerase (Invitrogen), which have low commercial price; and (iii) Advantage 2 Polymerase Mix (Clontech), a DNA polymerase with high commercial price. Herein, neither samples nor positive control amplified in the reactions carried out with Paq5000 DNA Polymerase. Furthermore, the reactions performed with Platinum® Taq DNA Polymerase showed contrasting results, amplifying alleles only in some samples. On the other hand, the two microsatellite loci used (amelogenin e D18S51) were amplified in the analyses carried out with the Advantage 2 Polymerase Mix. Our findings indicated that the enzymes Paq5000 DNA Polymerase and Platinum® Taq DNA Polymerase have low processing capacity, showing inefficiency in the microsatellite detection; and that Advantage 2 Polymerase Mix is highly effective and suitable for microsatellite amplification assays.
Dengue is an arboviral disease of great clinical relevance among neglected diseases. It is transmitted by Dengue virus (DENV), which infects about 25 million people worldwide each year. So far there is no available licensed vaccine or specific drug to be used against DENV, but only palliative treatments to overcome the symptoms of the disease, considered a serious public health problem. Among the molecular targets verified as strategies for developing antivirals against DENV, we highlight the serine proteasic complex NS2B-NS3pro, essential for the viral maturation and replication. This molecular target is found conserved in all four serotypes, acting in major viral processes. In this context, this study aimed to understand various aspects related to DENV and especially the proteasic complex NS2B-NS3pro as a catalytic mechanism and candidate inhibitors of this pharmacological target applied to dengue.
The entomopathogenic fungus Beauveria bassiana CG432 produced an extracellular lipase on the 7 th day of fermentation in Alves broth containing 0.25% Triton X-100 and Tween 80 surfactants, and 3.0% olive oil at 28°C and 200rpm. Optimal temperature and pH conditions for lipase activity were found to be range from 40 to 60°C and pH 7.5 and 8.0 using 50 mM Tris-HCl buffer, verified by a hydrolysis p -nitrophenol palmitate assay. Under these conditions, lipase was 50% stable for approximately 10h, however it was stable for up to 10 days (-18°C), refrigerated (4°C) or kept at room temperature (25°C).
The environment diversity is directly related to environmental balance, which is directly affected by industrial and municipal waste release. Some species of microorganisms can act as decomposers in bioremediation process, aiding in contaminated areas recovery. The Maringá city, located in state of Paraná – Brazil, have been demonstrated rapid population growth and excelling in the construction sector, textile, and others, resulting in an increase of waste generated, necessitating the deployment of new technologies for waste treatment, in order to reduce them, so that decrease the socio-economic and environmental impact. The city counts with the Plan for Waste Manager (PGR, by its initials in Portuguese) that aims at controlling the waste produced and the proper disposal thereof. Aiming to improve the treatment of waste generated, we made a survey to relate the industrial and domestic waste produced in Maringá with possible ways of treatment, in sustainable processes, using the bioremediation potential of microorganisms.
This study aimed to compare several DNA polymerases in order to optimize costs of microsatellite amplification reaction through the PowerPlex®16 BIO System. Three different polymerase brands have been used: (i) Paq5000 DNA Polymerase (Stratagene) and (ii) Platinum® Taq DNA Polymerase (Invitrogen), which have low commercial price; and (iii) Advantage 2 Polymerase Mix (Clontech), a DNA polymerase with high commercial price. Herein, neither samples nor positive control amplified in the reactions carried out with Paq5000 DNA Polymerase. Furthermore, the reactions performed with Platinum® Taq DNA Polymerase showed contrasting results, amplifying alleles only in some samples. On the other hand, the two microsatellite loci used (amelogenin e D18S51) were amplified in the analyses carried out with the Advantage 2 Polymerase Mix. Our findings indicated that the enzymes Paq5000 DNA Polymerase and Platinum® Taq DNA Polymerase have low processing capacity, showing inefficiency in the microsatellite detection; and that Advantage 2 Polymerase Mix is highly effective and suitable for microsatellite amplification assays
Humans need amino acids, especially the essential ones, which are obtained from the proteins ingested in the diet. With the rampant growth of the world population, it is necessary to increase the protein supplies, especially those of plant origin, which now play a major role as a food source. Leaves of ora-pronobis (Pereskia aculeata Miller) are a high quality protein source and have been used in the traditional cuisine of Minas Gerais for decades. However, it is important to analyze the anti-nutritional and/or toxic factors to consider the use of the plant leaves as an alternative source of nutrients. This work aimed to study the presence of proteic anti-nutritional factors, such as protease inhibitors and lectins, and to evaluate the “in vitro” digestibility throughout Tricine SDS-PAGE of these factors extracted from ora-pro-nobis (Pereskia aculeata Miller) leaves. The extraction of protease inhibitors and lectins from ora-pro-nobis leaves was performed in phosphate buffer. The partial purification of the fractions presenting activity of trypsin inhibitor and lectins was performed by ammonium sulfate precipitation and gel-filtration chromatography. The “in vitro” activity of the fractions with protease inhibitor and lectins, with or without heat treatment in the presence of digestive enzymes, showed that the cooking time of 1 minute was sufficient for protease inhibitors degradation. However, this time was not satisfactory for lectins, which remained resistant even after heating. This work shows that with proper cooking, ora-pro-nobis leaves can be used in the diet as a source of aminoacids
Conditions of carbonic anhydrase extraction and activity measurement should be adapted to plant species and to tissues of plants. Different extraction and reaction media have been compared in ten different tree species in order to define the best conditions for carbonic anhydrase activity measurement. Bias and variability of differences in measured values were analysed according to Bland and Altman method. A comparison of titrimetric method with the traditional electrometric method of estimation of CA activity in the ten species revealed that the measures were in agreement justifying the use of both methods for measurement of the enzyme activity interchangeably. Use of Tris-HCl buffer, 0.1 M (pH 8.3) allowed for a better stabilization of the enzyme during extraction and estimation. Use of potassium phosphate, sodium borate and veronal buffers is discouraged during extraction of the enzyme as the enzyme activities varied with species. Titrimetric method was found to be better over the electrometric method since change in colour appeared to be a better indicator of the end point of the reaction than measurements using a pH meter.
Citrus juice for export has a slight bitter taste which is characteristic and acceptable by the consumer but high levels of naringin reduce its quality and commercial value. Naringin (a flavonoid linked to a rhamnose and glucose) is the main bitter component of several citrus fruits. The reduction in bitterness may be achieved by naringinase, an enzyme that degrades naringin. This study aimed the production of naringinase of Aspergillus niger by fermentation of low-cost substrates from agroindustry. The experiment was planned according to a 2³ factorial design which included three replications at the center point having as variables (g/L): naringin (0.20; 0.30 and 0.40); soy bean molasses (0.35; 1.05 and 1.75 of total sugars) and sugar cane molasses (0.35; 1.05 and 1.75 of total sugars). Using the results of 144 hours of fermentation the statistical analysis and predictions were carried out. The statistical analysis showed that the significant variables for the production of naringinase were sugar cane molasses and soy molasses, as long as naringin was used at its lowest concentration. The predictive analysis showed a maximum value of 1.35U/mL could be achieved when naringin (0.25g/L), sugar cane molasses (1.75g/L) and soy bean molasses (1.75g/L) were used. When the validity test was carried out a value of 1.344 (99.3% of prevision) was obtained.
The objective of this study was to evaluate the chemical composition and technological properties of grains of soybean (Glycine max) BRS 284 and BMX Potência RR cultivars, produzed in Maua da Serra - PR, in the crop years 2011 and 2012. The proximate composition, alimentary fiber, isoflavones and technological properties were determined. Both cultivars showed high content of proteins BRS 284 (33,24%) and BMX Potência RR (34,74%), lipids BRS 284 (22,54%) and BMX Potência RR (21,72%) and total fiber BRS 284 (26,64%) and BMX Potência RR (27,13%), being the bigger fraction constituted of insoluble fiber. Both cultivars showed high value of total isoflavones BRS 284 (591.70 mg.100g-1) and BMX Potência RR (865.36 mg.100g-1), due to local cultivation conditions having lower averages temperatures. The BRS 284 and BMX Potência RR values for cultivars showed swelling volume (VI) of 4.31 mL.g-1 and 4.12 mL.g-1, water absorption index (IAA) of 2.75 g.g-1 and 2.75 g.g-1 and oil absorption index (IAO) of 2.87 g.g-1 and 2.60 g.g-1, respectively. The cultivars were different for moisture, lipids, protein, and total isoflavones. Both cultivars display characteristics to be used as raw material for various soybean foods.
A Ciclomaltodextrina-Glucanotransferase (CGTase) e a unica enzima capaz de converter amido em ciclodextrinas (CDs) pela reacao de ciclizacao das cadeias lineares. Ciclodextrinas sao oligossacarideos ciclicos contendo um minimo de 6 unidades de D-(+)-glicopiranose unidas por ligacoes alfa-1,4. Possuem capacidade de formar complexos de inclusao com grande variedade de moleculas hospedes em solucao. Com o objetivo exploratorio de aumentar a producao da CGTase para obtencao de CDs, realizou-se cultivo de Bacillus firmus CEPA 37, em reator batelada com pulsos de amido durante 48, 96 e 200 horas de cultivo. Observou-se a manutencao do pH entre 8,9 e 10, houve um bom consumo de substrato e um bom crescimento celular (em torno de 8 g/L). A atividade enzimatica de CGTase atingiu um pico de 0,11 U/mL apos 59 e 200 horas de cultivo, bem como uma atividade especifica com relacao a gama-CD de 1,4 U/mg de proteinas com 200 horas de cultivo. Houve aumento da quantidade de proteinas soluveis no meio de cultura apos os pulsos de amido, indicando que o micro-organismo buscou formas de disponibilizar nitrogenio para si, uma vez que a fonte de carbono foi suplementada e a de nitrogenio nao. Os Pulsos de amido tambem favoreceram o crescimento celular.
Humans need amino acids, especially the essential ones, which are obtained from the proteins ingested in the diet. With the rampant growth of the world population, it is necessary to increase the protein supplies, especially those of plant origin, which now play a major role as a food source. Leaves of ora-pro-nobis (Pereskia aculeata Miller) are a high quality protein source and have been used in the traditional cuisine of Minas Gerais for decades. However, it is important to analyze the anti-nutritional and/or toxic factors to consider the use of the plant leaves as an alternative source of nutrients. This work aimed to study the presence of proteic anti-nutritional factors, such as protease inhibitors and lectins, and to evaluate the “in vitro” digestibility throughout Tricine SDS-PAGE of these factors extracted from ora-pro-nobis (Pereskia aculeata Miller) leaves. The extraction of protease inhibitors and lectins from ora-pro-nobis leaves was performed in phosphate buffer. The partial purification of the fractions presenting activity of trypsin inhibitor and lectins was performed by ammonium sulfate precipitation and gel-filtration chromatography. The “in vitro” activity of the fractions with protease inhibitor and lectins, with or without heat treatment in the presence of digestive enzymes, showed that the cooking time of 1 minute was sufficient for protease inhibitors degradation. However, this time was notsatisfactory for lectins, which remained resistant even after heating. This work shows that with proper cooking, ora-pro-nobis leaves can be used in the diet as a source of aminoacids.
Strawberry is a fruit with fragile structure and high respiration rate, which results in a relatively short postharvest. With the increasing cultivation in Brazil, it is mainly consumed as fresh fruit or processed as jam or frozen pulp. Also an option for the processing of fermented fruit is strawberry. This study aimed to develop a manufacturing process of fermented strawberry in laboratory, and analyze the kinetics of fermentation beverage produced as well as its sensory characteristics. The drink had produced a delicate flavor with mild strawberry flavor, pH of 3.51, total acidity of 4.5 and an content of 9.62% being in conformity within the standards required by the Brazilian legislation on drinks.
The entomopathogenic fungus Beauveria bassiana penetrates the cuticle of insects by action of extracellular hydrolytic enzymes such as proteases, lipases and chitinases. Proteases can be induced in cultivation submerged in presence of protein nature. Thus, the objective of this work is to evaluate the production of proteases of B. bassiana CG432 in cultivation submerged in minimal medium Vogel supplemented with cuticle cicadas, Aedes aegypti , casein, chitin, gelatin, urea and amino acids: methionine, proline, alanine and glycine as substrates inductors. The inoculum contained 10 6 spores and cultivation was conducted to 28°C, 200 rpm for 5 days. Enzymatic extracts obtained by centrifugation were analyzed for protein content, production biomass and protease activity. The substrates examined in cultivation, the cuticle of cicadas, the larvae of A. aegypti and casein showed higher protease activity and, among the amino acids, methionine was presented the best result. Among these inducers to cuticle and the larvae were that induced higher specific activity enzyme and casein contributed to the higher biomass fungal growth.
A água é considerada o composto mais abundante, de suma importância e de incontáveis aplicações para o ser humano, como no abastecimento público, industrial ou agropecuário, preservação da vida aquática, recreação e transporte. No Brasil, soluções alternativas coletivas/SAC’s, como minas de água e poços artesianos ainda são utilizadas para abastecimento. Apesar de consideradas “limpas”, estas águas, utilizadas para consumo humano podem ser contaminadas durante o processo de captação ou durante seu trajeto até o ponto de consumo representando um risco à saúde coletiva devido à predisposição a doenças de veiculação hídrica. Neste trabalho foi avaliada, durante cinco meses, a qualidade da água de amostras de sete locais (SAC’s) da área urbana do município de Divinópolis - MG, por meio de análises físico-química, microbiológica e parasitológica. Foi detectada presença recorrente de coliformes fecais em dois locais, ausência de parasitas nas amostras analisadas e uma concentração significativa de nitrato em alguns pontos.
A imobilização de enzimas é uma alternativa para sua reutilização e redução de custos em processos industriais. O bagaço de cana-de-açúcar pode ser um suporte viável, pois é atóxico e tem baixo custo. A beta-glicosidase hidrolisa ligações beta-glicosídicas e pode ser aplicada em vários setores. Assim, o objetivo deste trabalho foi avaliar o bagaço de cana-de-açúcar submetido a tratamentos químicos e térmico para imobilização de beta-glicosidase de soja. O bagaço foi tratado com NaOH, etanol 70%, calor úmido a 121 ºC por 15 min e ativação com glutaraldeído 2,5%. A beta-glicosidase de soja foi obtida por ultrafiltração em membranas de 100 e 10 kDa. Os maiores rendimentos de imobilização da beta-glicosidase (68,3%) foram obtidos utilizando bagaço ativado com glutaraldeído e quando esta ativação foi precedida pelos tratamentos com NaOH e autoclavagem (76,1%). As médias de proteína e enzima imobilizada obtidas pelos tratamentos do bagaço não diferiram significativamente entre si (p?0,05).
As proteinas MYB figuram como uma das maiores familias de fatores de transcricao (FT) presentes em plantas, nas quais desempenham a regulacao de importantes processos, incluindo a defesa. A classe R2R3-MYB e a mais numerosa entre as plantas e a analise da conservacao do seu dominio MYB e seus motivos (C-terminal) permite agrupa-los em subgrupos. Estudos apontam que R2R3 estao envolvidas no controle de processos especificos, incluindo respostas a estresse biotico. Este trabalho objetivou a identificacao e classificacao filogenetica de R2R3-MYBs de soja, atraves de bioinformatica, bem como a analise de expressao dos FTs. A analise de 1390 sequencias proteicas permitiu a identificacao de 264 R2R3-MYBs, classificados em 42 subgrupos, assim como a atribuicao de funcoes putativas dos mesmos. Analise in silico do perfil de expressao dos MYBs (Genevestigator) demonstrou que eles sao induzidos por infeccao por Phakopsora pachyrhizi, tornando-os interessantes alvos para estudos funcionais futuros em soja. Palavras-chave : GmMYB, motivo, filogenia, P. pachyrhizi , soja
Visando reduzir custos com adubação nitrogenada e considerando o grande potencial biotecnológico dos microrganismos de solo, o objetivo deste trabalho foi estudar a diversidade genética de rizobactérias obtidas de distintas classes de solos da região oeste do Paraná cultivados sob diferentes manejos. Foram isoladas 400 estirpes, destas 269 foram submetidas à reação de Rep-PCR e destas, selecionou-se 103 estirpes para análise polifásica do perfil de PCR-RFLP da região que codifica o gene 16S rRNA. A biodiversidade encontrada pela análise estabeleceu um total de 10 agrupamentos genéticos, dos quais determinados manejos agrícolas, como o cultivo de crotalária, contribuíram com uma maior abundância de isolados. Os resultados demonstraram uma alta diversidade em todas as regiões avaliadas, apontando a influência concomitante dos diferentes manejos de cultivo e tipo de vegetação sobre a dominância de determinados grupos genéticos dentro das comunidades.
