Bioquímica e Biologia Molecular
URI permanente desta comunidadehttps://locus.ufv.br/handle/123456789/11837
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Item Elaboration and characterization of Japanese Raisin Tree (Hovenia dulcis Thumb.) pseudofruits fermented alcoholic beverage(Food Science and Technology, 2017-12) Pinto, Juliana Tensol; Oliveira, Tânia Toledo de; Queiroz, José Humberto de; Dias, Disney Ribeiro; Schwan, Rosane Freitas; Oliveira, Diego Pinto de; Alvarenga, Luana FarahHovenia dulcis pseudofruits have underexplored properties for food purposes, despite their pleasant sensory characteristics and therapeutic benefits. The aim of this study was the elaboration and chemical characterization of the alcoholic fermented beverage of H. dulcis, using selected strain of Saccharomyces cerevisiae (CCMA 0200). The resulting fermented beverage presented high content of phenolic compounds and antioxidant activity when compared to other fruits and beverages (DPPH and ABTS assay). The alcohol content was 12.9 oGL and total sugars 3.57g/L. By the GC-MS analysis, 39 compounds were identified including metabolites with therapeutic potential such as eugenol, trans-farnesol salicylates. The flavonoid dihidromyricetin was identified and quantified (75.17 mg/L) by HPLC-DAD and UPLC-MS/MS. The results reinforce the interest on nutraceutical and functional properties of this beverage and opens perspectives for new studies that value this underexplored pseudofruit.Item Hydrolysis of galacto-oligosaccharides in soy molasses by α -galactosidases and invertase from Aspergillus terreus(Brazilian Archives of Biology and Technology, 2010-05) Reis, Angélica Pataro; Guimarães, Valéria Monteze; Ferreira, Joana Gasperazzo; Queiroz, José Humberto de; Oliveira, Maria Goreti Almeida; Falkoski, Daniel Luciano; Almeida, Maíra Nicolau de; Rezende, Sebastião Tavares deTwo α -galactosidase (P1 and P2) and one invertase present in the culture of Aspergillus terreus grown on wheat straw for 168 h at 28ºC were partially purified by gel filtration and hydrophobic interaction chromatographies. Optimum pH and temperatures for P1, P2 and invertase preparations were 4.5-5.0, 5.5 and 4.0 and 60, 55 and 65ºC, respectively. The KM app for ρ -nitrophenyl-α -D-galactopyranoside were 1.32 mM and 0.72 mM for P1 and P2, respectively, while the KM app value for invertase, using sacarose as a substrate was 15.66 mM. Enzyme preparations P1 and P2 maintained their activities after pre-incubation for 3 h at 50ºC and invertase maintained about 90% after 6 h at 55 ºC. P1 and P2 presented different inhibition sensitivities by Ag+, D-galactose, and SDS. All enzyme preparations hydrolyzed galacto-ologosaccharides present in soymolasses.Item Nematicidal activity of Paecilomyces marquandii proteases on infective larvae of Ancylostoma spp(Brazilian Archives of Biology and Technology, 2016-01) Genier, Hugo Leonardo André; Queiroz, José Humberto de; Braga, Fabio Ribeiro; Soares, Filippe Elias de Freitas; Araújo, Jackson Victor de; Pinheiro, Iara RebouçasThe present study aimed to evaluate the action of Paecilomyces marquandii proteases on Ancylostoma spp L3. White halos in the zymogram confirmed the proteolytic action. Difference (p <0.01) between the number of L3 in the differents groups was found, with 41.4% of reduction of Ancylostoma spp. L3 before 24 hours.Item Perfil biométrico, histomorfométrico e bioquímico no tratamento com atorvastatina cálcica de ratas com osteoporose induzida com dexametasona(Revista Brasileira de Ortopedia (English Edition), 2018-09) Ferreira Junior, Davilson Bragine; Pizziolo, Virgínia Ramos; Oliveira, Tânia Toledo de; Matta, Sérgio Luis Pinto da; Píccolo, Mayra Soares; Queiroz, José Humberto deAvaliar os efeitos da atorvastatina cálcica no tratamento da osteoporose induzida com dexametasona. A indução da osteoporose consistiu na administração de dexametasona na dose de 7,5 mg/kg de peso corporal, por via intramuscular, uma vez por semana durante quatro semanas, à exceção dos animais do grupo controle (G1). Os animais foram distribuídos nos seguintes grupos: G1 (grupo controle sem osteoporose), G2 (grupo controle com osteoporose sem tratamento), G3 (grupo controle com osteoporose tratado com alendronato de sódio 0,2 mg/kg) e G4 (grupo com osteoporose tratado com atorvastatina cálcica 1,2 mg/kg). Após 30 e 60 dias do início do tratamento dos animais, foram feitas as dosagens dos níveis séricos de fosfatase alcalina, fosfatase alcalina óssea, avaliação biométrica e histomorfométrica óssea. Em relação às análises biométricas e histomorfométricas, aos 60 dias de tratamento o G4 apresentou densidade óssea (índice Seedor), densidade trabecular óssea e espessura da cortical de 0,222 ± 0,004 g/cm, 59,167 ± 2,401% e 387,501 ± 8,573 μm, respectivamente, com diferença positiva, estatisticamente significativa (p < 0,05), em relação ao grupo G2. Aos 30 e 60 dias de tratamento, o G4 apresentou níveis séricos de fosfatase alcalina óssea estatisticamente significativos (p < 0,05) e superiores a todos os grupos (7,451 ± 0,173 μg/L e 7,473 ± 0,529 μg/L, respectivamente). O tratamento com atorvastatina cálcica demonstrou a capacidade desse fármaco de aumentar a atividade osteoblástica e a atividade reparadora tecidual óssea, atuar de forma diferente do alendronato de sódio, que demonstrou atividade preponderantemente antirreabsortiva.Item In vitro and in silico characterization of a novel dextranase from Pochonia chlamydosporia(3 Biotech, 2018-03) Sufiate, Bruna; Araújo, Jackson Victor de; Queiroz, José Humberto de; Gouveia, Angélica S; Cardoso, Evandro; Moreira, Samara Silveira; Soares, Filippe Elias de Freitas; Braga, Fabio RibeiroThe objective of this study was to purify, characterize, and phylogenetically and structurally analyze the dextranase produced by the fungus Pochonia chlamydosporia. Dextranase produced by the fungus P. chlamydosporia was purified to homogeneity in two steps, with a yield of 152%, purification factor of 6.84 and specific activity of 358.63 U/mg. Its molecular weight was estimated by SDS-PAGE at 64 kDa. The enzyme presented higher activity at 50 °C and pH 5.0, using 100 mM citrate–phosphate buffer, was inhibited by Ag1+, Hg2+, Cu2+, Mg2+, and presented KM of 23.60 µM. Mature dextranase is composed of 585 amino acids residues, with a predicted molecular weight of 64.38 kDa and pI 5.96. This dextranase showed a strong phylogenetic similarity when compared to Trichoderma harzianum dextranase. Its structure consists of two domains: the first composed by 15 β strands, and the second composed by a right-handed parallel β-helix.Item Minimum cocktail of cellulolytic multi-enzyme complexes obtained from white rot fungi via solid-state fermentation(3 Biotech, 2018-01) Soares, Filippe Elias de Freitas; Queiroz, Paula Viana; Tavares, Gabriella Peterlini; Sufiate, Bruna Leite; Queiroz, José Humberto de; Cardoso, Wilton Soares; Santos, Fernando Almeida; Kasuya, Maria Catarina MegumiThe objective of this work was to optimize the total cellulase activity of the crude extract cocktails from five white rot fungi produced by solid-state fermentation, by means of the central composite design. The white rot fungi Pleurotus ostreatus PLO 06, Pleurotus eryngii PLE 04, Trametes versicolor TRAM 01, Pycnosporus sanguineus PYC 02 and Phanerochaete chrysosporium PC were tested. For optimization process aiming at the maximum value of total cellulase activity (FPAse), the multi-enzyme cellulase complexes (crude extracts) of each fungus were mixed simultaneously in different proportions. There was increase in FPAse activity for the cocktails formed by the extracts of the five fungi together, compared to the extracts of each fungus alone. The model presented the minimum cocktail of enzymes for maximum total cellulase activity, with 100.00 μL PYC; 100.00 μL PC; 100.00 μL PLO06; 100.00 μL PLE04 and 200 μL TRAM01. The maximum value found was of 304.86 U/L. The result of the cocktails was very relevant, showing that there is an enzymatic complementation in the extracts that should be further studied. Concentrated extract cocktails should also be evaluated for biomass saccharification.Item Nematicidal action of Pleurotus eryngii metabolites(Biocatalysis and Agricultural Biotechnology, 2017-10) Sufiate, Bruna Leite; Soares, Filippe Elias de Freitas; Moreira, Samara Silveira; Gouveia, Angélica de Souza; Monteiro, Thalita Suelen Avelar; Freitas, Leandro Grassi de Freita; Queiroz, José Humberto dePleurotus eryngii is one of the most cultivated and consumed mushroom species in North Africa, Europe and Asia. Fungi from Pleurotus genus have demonstrated nematophagous activities, however most of the literature reports are focused on Pleurotus ostreatus. The aim of this work was to evaluate P. eryngii action on Panagrellus sp. and to evaluate the effect of this fungus culture extract on Meloidogyne javanica eggs. P. eryngii fungus and its extract significantly reduced (p < 0.01) the number of intact Panagrellus sp. larvae after 24 h treatment in 60% and 90%, respectively. This effect is not related to enzymatic activity, but to the presence of other metabolites. M. javanica eggs, when treated with P. eryngii extract, showed a 53% reduction (p < 0.01) in the number of intact eggs. The M. javanica intact eggs reduction is attributed to enzymatic activity, once the extract showed proteolytic and chitinolytic activities of, respectively, 32.74 and 3.57 U/mL. P. eryngii fungus has predatory activity against Panagrellus sp. larvae due to toxins production and negatively affects M. javanica eggs and juveniles development due to chitinases and proteases production, evidencing this fungus potential to be used in biological control.Item Nematicidal activity of proteases from Euphorbia milii(Biocatalysis and Agricultural Biotechnology, 2017-04) Sufiate, Bruna Leite; Soares, Filippe Elias de Freitas; Freitas, Filippe Elias de; Queiroz, José Humberto deThe present work aimed to characterize the proteases present in Euphorbia milii latex and to evaluate the nematicidal potential of these enzymes. Results suggest that proteases present in E. milii latex concentrate samples are milin and miliin. Difference (p < 0.01) between larvae number in the different groups was found, with 65.59% and 96.46% Panagrellus redivivus larvae reduction after 24 and 48 h, respectively.Item The nematophagous fungus Monacrosporium thaumasium and its nematicidal activity on Angiostrongylus vasorum(Revista Iberoamericana de Micología, 2013-09-24) Soares, Filippe Elias de Freitas; Braga, Fabio Ribeiro; Araújo, Jackson Victor de; Lima, Walter dos Santos; Queiroz, José Humberto deThe dog acts as a reservoir and environmental disseminator of potentially zoonotic parasites. The objective of this work was to study the fungus Monacrosporium thaumasium regarding its nematicidal potential in laboratory trials and its proteolytic profile. The in vitro test was carried out through two assays (A and B). In assay A, conidia of the fungus N34a were added in positive coprocultures for Angiostrongylus vasorum. In assay B, crude extract (treated group) and distilled water (control group) were added to coprocultures. Next, the proteolytic profile of crude extract of the nematophagous fungus M. thaumasium (NF34a) was revealed by performing a zymogram. There was a reduction (p < 0.01) in the averages of larvae recovered from the treated groups (conidia and crude extract) in relation to control groups. The zymogram suggested that the nematophagous fungus M. thaumasium produces a protease of approximately 40 kDa. The results of this work confirm that the conidia as well as the crude extract of the fungus M. thaumasium may be used to control A. vasorum L1. The proteolytic profile suggested the presence of one protease (Mt1) of approximately 40 kDa that in the future may be used in biological control of L1 of this nematode.Item Use of sorghum straw (Sorghum bicolor) for second generation ethanol production: pretreatment and enzymatic hydrolysis(Química Nova, 2013-04-04) Tavares, Gabriella Peterlini; Cardoso, Wilton Soares; Tardin, Flávio Dessaune; Queiroz, Paula Viana; Mota, Samuel Sampaio; Kasuya, Maria Catarina Megumi; Queiroz, José Humberto deAgronomic biomass yields of forage sorghum BRS 655 presented similar results to other energy crops, producing 9 to 12.6 tons/ha (dry mass) of sorghum straw. The objective of this study was to evaluate the lignocellulosic part of this cultivar in terms of its potential in the different unit processes in the production of cellulosic ethanol, measuring the effects of pretreatment and enzymatic hydrolysis. Three types of pre-treatments for two reaction times were conducted to evaluate the characteristics of the pulp for subsequent saccharification. The pulp pretreated by alkali, and by acid followed by delignification, attained hydrolysis rates of over 90%.