ABSTRACT
This study demonstrates the production of an active enzyme cocktail produced by growing Trichoderma harzianum on sugarcane bagasse. The component enzymes were identified by LCMS-MS. Glycosyl hydrolases were the most abundant class of proteins, representing 67% of total secreted protein. Other carbohydrate active enzymes involved in cell wall deconstruction included lytic polysaccharide mono-oxygenases (AA9), carbohydrate-binding modules, carbohydrate esterases and swollenin, all present at levels of 1%. In total, proteases and lipases represented 5 and 1% of the total secretome, respectively, with the rest of the secretome being made up of proteins of unknown or putative function. This enzyme cocktail was efficient in catalysing the hydrolysis of sugarcane bagasse cellulolignin to fermentable sugars for potential use in ethanol production. Apart from mapping the secretome of T. harzianum, which is a very important tool to understand the catalytic performance of enzyme cocktails, the gene coding for T. harzianum swollenin was expressed in Aspergillus niger. This novel aspect in this work, allowed increasing the swollenin concentration by 95 fold. This is the first report about the heterologous expression of swollenin from T. harzianum, and the findings are of interest in enriching enzyme cocktail with this important accessory protein which takes part in the cellulose amorphogenesis. Despite lacking detectable glycoside activity, the addition of swollenin of T. harzianum increased by two-fold the hydrolysis efficiency of a commercial cellulase cocktail. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:327-336, 2016.
Subject(s)
Cellulases/analysis , Cellulose/metabolism , Proteome/metabolism , Saccharum/metabolism , Trichoderma/metabolism , Biocatalysis , Cellulases/metabolism , Cellulose/biosynthesis , Cellulose/chemistry , Hydrolysis , Proteome/chemistry , Saccharum/chemistry , Trichoderma/chemistryABSTRACT
The present study validates the methodology and the information collecting instrument for analysis of nutritional security/insecurity on the urban and rural family level, proposed by the USDA (United States Department of Agriculture). An intentional sample of domiciles was selected to represent different social strata, 194 families were enrolled in urban Manaus, of which 174 had children, in the following neighborhoods: Jesus me Deu, Nova Israel, Cidade Nova, Coroado and Conj. Petro. In the rural area between the municipalities of Iranduba and Manacapuru, 209 riparian families were interviewed, and of these 131 had children. The final validation of the questionnaire (global internal consistency) was made by comparing, the levels of nutritional security/insecurity, with the defined social strata and food consumption indicators. The results demonstrated that the groups of highest nutritional insecurity were the very poor. The instrument presented high validation and internal consistency.
O presente estudo validou a metodologia e o instrumento de coleta de informação para análise da segurança/insegurança alimentar, em famílias urbanas e rurais no estado do Amazonas conforme o proposto pelo USDA (Departamento de Agricultura dos Estados Unidos). Valendo-se de amostra intencional de domicílios, selecionadas para representar estratos sociais diferentes foram computadas 194 famílias sendo 174 com crianças na área urbana de Manaus, envolvendo os seguintes bairros: Jesus me Deu, Novo Israel, Cidade Nova, Coroado e Conjunto Petro. Na área rural foram entrevistadas 209 famílias ribeirinhas e destas 131 com crianças, distribuídas entre os Municípios de Iranduba e Manacapuru. A validação final do questionário (Consistência interna global) deu-se por meio da comparação dos níveis de segurança e insegurança alimentar, com os estratos definidos dos indicadores sociais e de consumo. Pode-se concluir que os grupos com maior insegurança alimentar foram os situados em estratos sociais mais baixos e de baixo consumo de alimentos sensíveis a estas condições. O instrumento de coleta apresentou alta validade e consistência interna.