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Master's Dissertation
DOI
10.11606/D.87.2012.tde-06062012-102804
Document
Author
Full name
Felipe Senne de Oliveira Lino
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2012
Supervisor
Committee
Kilikian, Beatriz Vahan (President)
Azzoni, Adriano Rodrigues
Gomez, José Gregorio Cabrera
Title in Portuguese
Engenharia evolutiva aplicada a Trichoderma sp. para produção de celulases.
Keywords in Portuguese
Trichoderma
Engenharia de produção
Microbiologia industrial
Mutagênese
Abstract in Portuguese
O projeto visou aumentar a produção de celulases em fungos T. harzianum IPT 821 e T. reesei QM 9414. Esporos foram submetidos à radiação UV-C. Células das colônias mais capacitadas ao crescimento foram sucessivamente cultivadas em meio solidificado, contendo concentrações progressivamente reduzidas fonte de carbono, de modo a resultar pressão ambiental seletiva e crescente. Após esta etapa as linhagens isoladas foram cultivadas em meio composto por bagaço de cana/farelo de trigo na proporção 80/20, 60% de umidade, 30°C por 72h. Uma linhagem, originária da cepa IPT 821, apresentou atividade de 3,7 FP U/gms, 50% superior em relação à parental: 2,6 U/gms (p=0,001; p<0,05). Análises das frações enzimáticas indicaram uma diferença significativa (p=0,001; p<0,05) na atividade de xilanase: 4,7 U/gms (mutante) e 4 U/gms (parental). Ensaios de hidrólise, Avicel como substrato (1% de sólidos; w/v) indicaram um aumento de quase 70% na hidrólise em 48h, da mutante em comparação à parental (8,7% (mutante) e 4,8% (parental), concentração enzimática de 2,5 FP U/gms).
Title in English
Evolutionary engineering applied in Trichoderma sp. for the production of cellulase.
Keywords in English
Trichoderma
Industrial microbiology
Mutagenesis
Production engineering
Abstract in English
This project aimed to increase cellulase production in fungi T. harzianum IPT 821 and T. reesei QM 9414. Spores were exposed to UV-C radiation. Colonies were plated and those showing best growth were successively cultivated in plates containing increasingly stress conditions reduced concentrations of the carbon source thus creating a progressive selective pressure. After this pre-selection step isolated strains were cultivated in medium comprising sugar cane bagasse and wheat straw, in the proportion of 80/20 respectively, 60% of moisture, 30°C for 72h. One strain, originated from IPT 821 strain, showed a cellulolytic activity of 3,7 U/gdw; 50% superior to the parental strain: 2,6 U/gdw (p=0,001; p<0,05). Analysis of the enzymatic cocktail showed significant difference (p=0,001; for p <0,05) on xylanase activity: 4,7 U/gdw (mutant strain) and 4 U/gdw (parental strain). Hydrolysis assays, using Avicel as substrate (1% w/v) showed an increase on hydrolysis of about 70%, in 48h (8,7% (mutant strain) and 4,8% (parental strain), enzyme load of about 2,5 U/gdw).
 
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Publishing Date
2012-08-22
 
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