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Doctoral Thesis
DOI
https://doi.org/10.11606/T.87.2015.tde-04092015-151800
Document
Author
Full name
Diogo Robl
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2015
Supervisor
Committee
Araujo, Welington Luiz de (President)
Buckeridge, Marcos Silveira
Gomez, José Gregorio Cabrera
Piccoli, Rosane Aparecida Moniz
Taciro, Marilda Keico
Title in English
Hemicellulases and acessory proteins from filamentous fungi and actinomycetes for lignocellulose biomass deconstruction.
Keywords in English
Actinomycetes
Endophytics
Fungi
Hemicellulases
Sugar cane
Abstract in English
Endophytic microorganisms were screened for hemicellulases production using plate assays and liquid cultivations. Two strains were selected and used in further studies. Aspergillus niger DR02 strain and Annulohypoxylon stigyum DR47. In A.niger fed-batch submerged cultivation approaches were developed using liquor from hydrothermal sugar cane pretreatment, and maximum xylanase activities obtained were 458.1 U/mL for constant fed-batch mode. For A. stygium DR47 media optimization and bioreactor cultivation using citrus bagasse and soybean bran were explored and revealed a maximum production of 6.26 U/mL of pectinase and 10.13 U/mL of β-glucosidase. Improved hemicellulase production was also done by genetic modifications at carbon catabolic repression levels in A. niger. Mass spectrometric studies were done in enzymatic extracts produced and contributed to understand the enzymatic supplementation of Celluclast 1.5L.In addition the extracts produced were tested for an enzyme formulation and that was able to increase sugar cane bagasse hydrolysis.
Title in Portuguese
Hemicelulases e proteínas acessórias de fungos filamentosos e de actinomicetos para desconstrução de biomassa lignocelulósica.
Keywords in Portuguese
Actinomicetos
Cana de açúcar
Endofíticos
Fungos
Hemicelulases
Abstract in Portuguese
Microrganismos endofíticos foram selecionados para a produção de hemicelulases através de testes em placa e de cultivo líquido. Duas linhagens foram selecionadas para posteriores cultivos, Aspergillus niger DR02 e Annulohypoxylon stigyum DR47. Para o fungo A.niger batelada alimentada em cultivo submerso com licor do tratamento hidrotérmico do bagaço de cana de açúcar permitiram obtenção de máxima atividade de xilanase de 458,1 U/mL. Para a linhagem A. stygium DR47 otimização do cultivo utilizando bagaço cítrico e farelo de soja foram explorados e revelaram produção de 6,26 U/mL de pectinase e 10,13 U/mL de β-glucosidase. Melhoramento da produção de hemicelulase foi realizado com modificações genéticas ao nível de repressão catabólica de carbono em A. niger. Estudos de espectrometria de massas foram realizados nos extratos enzimáticos produzidos e contribuíram para entender a suplementação da Celluclast 1.5L. Além disso, os extratos produzidos foram testados para uma formulação que foi capaz de aumentar a hidrólise do bagaço de cana de açúcar.
 
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Publishing Date
2015-09-17
 
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