Análise de metilxantinas em dezesseis progênies de erva mate extraídas por dióxido de carbono supercrítico.

A Ilex paraguariensis apresenta grande potencial de utilização pela diversidade de seus compostos químicos. Dentre os principais grupos de compostos presentes em erva mate citam-se as metilxantinas, com predominância de cafeína, teobromina e traços de teofilina. Na quantificação convencional destes compostos utiliza-se extração com solvente orgânico que expressa o conteúdo total de cafeína presente na amostra. Como forma alternativa de obtenção das metilxantinas pode-se utilizar a extração por fluído supercrítico (EFSC). Esta extração caracteriza-se pela obtenção de produtos de elevada qualidade, sem as inconveniências de resíduos de solventes e alterações nas propriedades do extrato presente na extração convencional. O objetivo deste trabalho foi quantificar metilxantinas presentes nas amostras de dezesseis progênies oriundas de quatro procedências (Ivaí/PR, Barão de Cotegipe/RS, Quedas do Iguaçu/PR e Cascavel/PR), cultivadas em três localidades (Ivaí/PR, Rio Azul/PR e Guarapuava/PR) utilizando a extração por solventes orgânicos e pela EFSC usando como solvente o CO2. As metilxantinas foram quantificadas por HPLC e espectrometria comparando-se com padrão. Os teores médios de metilxantinas, correspondendo à soma de cafeína e teobromina, foram de 19,112 mg/100 g nas progênies de Ivaí/PR, 8,906 mg/100 g nas progênies de Rio Azul/PR e de 12,796 mg/100 g nas progênies de Guarapuava/PR. Ao compararmos a extração supercrítica com a extração convencional de cafeína os valores médios encontrados foram de 2,808±0,7 % para Ivaí/PR, 1,537±0,2 % em Rio Azul/PR e 1,728±0,5 % para Guarapuava/PR. A EFSC usando o dióxido de carbono como solvente aliada à análise HPLC mostrou-se eficiente na caracterização e quantificação das metilxantinas presentes nas amostras analisadas.Seção: Controle de Qualidade/ Composição Química. Feira do Agronegócio da Erva-mate, 1., 2003, Chapecó. Integrar para promover o agronegócio da erva-mate

Application of an endo-xylanase from Aspergillus japonicus in the fruit juice clarification and fruit peel waste hydrolysis.

The endo-xylanase from Aspergillus japonicus (UFMS 48.136) was purified in a single step using carboximethylcellulose chromatographic column and applied in fruit juice clarification process and fruit peel waste hydrolysis. This purification procedure resulted in 38.9-fold purification of endo-xylanase with 83.3% final yield. MALDITOF analysis confirmed the molecular mass of 32 kDa. The optimal purified endo-xylanase activity was at a range of pH from 5.0 to 6.0 and from 50 to 60 +-C, retaining more than 70% of its activity at all pH studied (3.0?8.0) for 24 h at room temperature. The A. japonicus endo-xylanolytic activity stimulation curve was assayed in the presence of different birchwood xylan concentrations (ranging from 0.02 to 0.5% w/v) and the endoxylanase activity presented a Vmax of 467.4 +- 30.38 μmol/min/mg, with a km of 2.59 +- 0.17 mg/mL, a kcat of 253.95 +- 16.51 s -1 and a kcat/km value of 98.05 +- 4.41 mL s -1 mg -1. The endo-xylanase was activated by Mn2þ (34.5%) and inhibited by Cu2þ (56.9%). The endo-xylanase was activated by β-mercaptoethanol, Triton X-100, Tween-20, Tween-80 and ferulic acid. In the clarification assay, endo-xylanase successfully clarified the juices of mango (51.11%), banana (9.99%) and tangerine (8.54%). Furthermore, the enzyme also hydrolysed all fruit peel wastes that were tested. In summary, A. japonicus endo-xylanase showed potential for applications in fruit juice clarification and in the treatment of fruit peel wastes, and it is a good candidate for the food industry due to its wide pH stability under acidic conditions

A novel xylan degrading β-D-xylosidase: purification and biochemical characterization

Aspergillus ochraceus, a thermotolerant fungus isolated in Brazil from decomposing materials, produced an extracellular b-xylosidase that was purified using DEAE-cellulose ion exchange chromatography, Sephadex G-100 and Biogel P-60 gel filtration. b-xylosidase is a glycoprotein (39 % carbohydrate content) and has a molecular mass of 137 kDa by SDS-PAGE, with optimal temperature and pH at 70 C and 3.0–5.5, respectively.b-xylosidase was stable in acidic pH (3.0–6.0) and 70 C for 1 h. The enzyme was activated by 5 mM MnCl2 (28 %)and MgCl2 (20 %) salts. The b-xylosidase produced by A. ochraceus preferentially hydrolyzed p-nitrophenyl-b- D-xylopyranoside, exhibiting apparent Km and Vmax values of 0.66 mM and 39 U (mg protein)-1 respectively, and to a lesser extent p-nitrophenyl-b-D-glucopyranoside. The enzyme was able to hydrolyze xylan from different sources,suggesting a novel b-D-xylosidase that degrades xylan. HPLC analysis revealed xylans of different compositions which allowed explaining the differences in specificity observed by b-xylosidase. TLC confirmed the capacity.This work was supported by the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), and the Conselho de Desenvolvimento Científico e Tecnológico (CNPq). J. A. J. and M. L. T. M. P are Research Fellows of CNPq. M. M. was a recipient of a FAPESP fellowship and this work is part of her Doctoral Thesis. It is also part of the project SISBIOTA CNPq: 563260/2010-6 and FAPESP: 2010/52322-3