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Doctoral Thesis
DOI
https://doi.org/10.11606/T.11.2019.tde-20191220-131804
Document
Author
Full name
Maria Bernadete Deléo Aguilar
Institute/School/College
Knowledge Area
Date of Defense
Published
Piracicaba, 1991
Supervisor
Title in Portuguese
Atividade celulolítica e protoplastização em Humicola sp e Trichoderma pseudokoningii var. Rifai
Keywords in Portuguese
ATIVIDADE ENZIMÁTICA
CELULASE
FUNGOS CELULOLÍTICOS
FUSÃO DE PROTOPLASTOS
MELHORAMENTO GENÉTICO
Abstract in Portuguese
No presente trabalho foram realizados estudos com as linhagens selvagem e mutante morfológico mor13 de Humicola e as linhagens selvagem e mutantes auxotróficos leu e ribo de Trichoderma pseudokoningii, através de utilização de técnicas de obtenção, regeneração e fusão de protoplastos, visando o melhoramento genético da produção de celulases, visando o melhoramento genético da produção de celulases. A análise de crescimento sob três temperaturas de incubação revelou que a temperatura de 37° C foi favorável tanto para as linhagens de Humicola sp como para as linhagens de Trichoderma pseudokoningii. Foram realizadas dosagens das atividades das enzimas avicelase, carboximeticelulase e β- glucosidase das cinco linhagens com a finalidade de comparar sua capacidade celulolítica. As atividades da avicelase e carboximetilcelulase foram altas e não houveram diferenças significativas entre as linhagens. As atividades da β-glucosidase entre as linhagens foram estatisticamente diferentes, sendo que a atividade do mutante mor13 de Humicola sp foi superior às atividades da β-glucosidases das demais linhagens. Os protoplastos das linhagens de Humicola sp foram obtidos após 1 hora de tratamento enzimático sendo o melhor estabilizador osmótico, o sulfato de magnésio; enquanto que para as linhagens de Trichoderma pseudokoningii, o tratamento enzimático foi de 3 horas e o melhor estabilizador osmótico foi o sulfato de amônia. A regeneração dos protoplastos de todas as linhagens dos dois gêneros foi conseguida pela utilização de uma mistura de sulfato de magnésio e sulfato de amônia. Através do uso de polietineloglicol e da técnica de eletrofusão, não foram obtidos produtos de fusão.
Title in English
Not Avaliable
Abstract in English
In this work studies were carried out with the wild and morphological mor13 mutant strains of Humicola sp and the wild, auxotropic leu, ribo mutants strains of Trichoderma pseudokoningii, aiming the genetic breeding for the cellulose production through the improvement of the techniques of obtention, regeneration and fusion of protoplasts. The growth analysis under three incubation temperatures revealed that the temperature of 37° C was favorable for the growth of the Humicola sp strains as much as the Trichoderma pseudokonigii strains. The dosages of the enzymes activities, endoglucanase, exoglucanase and β-glucosidase of the five strains were performed with the objective of to compare your cellulolytic ability. The endoglucanase and exoglucanase activities were high and no significative differences were detected among the strains. The β-glucosidase activities were different statistically among the strains and the activity of the mor13 mutant of Humicola sp was higher than the activities of β-glucosidases of the others strains. The protoplasts of the Humicola sp strains were obtained after one hour of enzymatic treatment, and the better osmotic stabilizier was magnesium sulphate; while that for the Trichoderma pseudokoningii strains, the enzymatic treatment was of three hours and the better osmotic stabilizer was ammonium sulphate. The regeneration of the protoplasts of each strain of the both genera was obtained by use of the mixture of magnesium sulphate with ammonium sulphate. Through the use of polyethylene glycol or of the electrofusion technique no fusion products was obtained.
 
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Publishing Date
2019-12-20
 
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