Produção, recuperação e caracterização de proteinas alergenicas da biomassa de Drechslera (Helminthosporium) monoceras obtida por fermentação em estado solido

AUTOR(ES)
DATA DE PUBLICAÇÃO

2002

RESUMO

The purpose of this work was the production, recovery and characterization of allergenic proteins of Drechslera (Helminthosporium) monoceras biomass; cultivated by solid-state fermentation, for the obtainment of standardized mould allergenic extract, applicable to diagnosis of allergy diseases. Bioreactors based on Raimbault-type aerated fixed bed columns were used for solid-state fermentation, and sugar cane bagasse, wheat and maize bran were tested as substrate. The most important variables of solid fermentation were optimized by statistical experimental design and by surface response analysis, and the best results, in terms of productivity, were obtained at pH 9.5 and 45.8% moisture. The solid state fermentation conditions were determined for wheat bran used as substrate (with 0.59 mm dimension); 0.4 mg/mL of inoculum; 2.0 Uh of air and 25°C. The kinetic of biomass growth was determined under the optimized conditions of the process. Water was used as solvent for the obtainment of the crude extracts. Potentially allergenic extracts were obtained by protein precipitation from the crude extracts by the use of organic solvents and ammonium sulfate, followed by salt and polyphenols removal. Characterization of potentially allergenic extracts was done by SDS-PAGE electrophoresis, isoeletric focusing, gel permeation chromatography and by cutaneous "prick-tests", dot-blotting and immunoblotting analysis for the allergenic evaluation of extracts. Twelve proteins were identified from the potentially allergenic extracts, in the range of low and high molecular mass, between 14 and 156 kDa. From cutaneous tests, 24% of the patients with respiratory allergy symptoms showed positive reactions. The immunoblotting assays revealed six proteins with allergenic activity due to their stronger reactivity, with the following molecular masses: 155.9; 104.2; 44.2; 35.7; 27.8 and 14.3 kDa

ASSUNTO(S)

engenharia bioquimica fermentação alergia fungos - extração de proteinas

ACESSO AO ARTIGO

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