Produção e avaliação das aplicações de enzimas quitinolíticas e queratinolíticas
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Data
2011-08-25
Autores
Santos, Emerson dos [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
Enzimas são catalisadores fundamentais para os sistemas biológicos sendo amplamente estudadas e aplicadas em todo o mundo, especialmente na área da saúde. Neste trabalho foi avaliada a aplicação de enzimas quitinolíticas e queratinolíticas em ensaios in vitro contra Pedicullus humanus capitis e na degradação de calos humanos, respectivamente. Foram selecionados os micro-organismos com maior potencial para produção dessas enzimas e determinados os parâmetros de produção, as condições de separação, purificação e caracterização das enzimas. Os micro-organismos com maior potencial para a produção de quitinases e queratinases foram os fungos Metarhizium anisopliae CG374 e Aspergillus oryzae, respectivamente. As maiores produções de quitinases foram obtidas em biorreator a 28°C, pH 7,0, 1,5 vvm e 200 rpm após 120 h. Quando produzida em biorreator hiperbárico sob pressão de 5 bar, apresentou um aumento de 21,3% na produtividade em relação ao biorreator convencional. Para queratinases em biorreator, os parâmetros de produção foram 28°C, pH 6, 200 rpm, 1,5 vvm e 120 h. A produção das quitinases e queratinases em biorreator foi 83 e 154% maior do que em frascos Erlenmeyer, respectivamente. A quitinase foi concentrada 5,12 vezes na purificação apresentando massas molares aproximadas de 24, 28 e 80 kDa. Fração purificada contendo quitinase apresentou estabilidade a 40°C por 60 minutos. A queratinase foi concentrada 4,52 vezes após purificação com massas molares de 40 e 50 kDa apresentando estabilidade térmica a 40°C por 60 min. A avaliação das enzimas quitinolíticas frente a piolhos humanos apontou para uma eficiência 9,1% maior quando comparadas com produtos comerciais. A avaliação da ação de enzimas queratinolíticas na degradação de calos humanos demonstrou resultados significativos quando comparados com produtos disponíveis no mercado...
Enzymes are catalysts for fundamental biological systems is extensively studied and applied worldwide, especially in health. In this study were evaluated the application of enzymes chitinolytic and keratinolytic in tests in vitro against Pedicullus humanus capitis and degradation of human callus, respectively. Were selected microorganisms with the greatest potential for production of these enzymes and determined the production parameters, conditions of separation, purification and characterization of enzymes. The microorganisms with the higher potential for the production of chitinases and keratinases were Metarhizium anisopliae CG374 and Aspergillus oryzae, respectively. The highest yields of chitinases obtained in fermentations in a bioreactor were 28°C, pH 7.0, 1.5 vvm and 200 rpm after 120 h. When produced in the hyperbaric bioreactor at a pressure of 5 bar, an increase of 21.3% in yield over conventional production in a conventional bioreactor was observed. To keratinases in a bioreactor, the production parameters were 28°C, pH 6.0, 200 rpm, and 1.5 vvm after 120h. The production of chitinases and keratinases in bioreactor was 83 and 154% higher than in Erlenmeyer flasks, respectively. The chitinase was 5.12 times concentred on the purification presenting molar weight of approximately 24, 28 and 78 kDa. Fraction containing purified chitinase showed thermal stability of 40°C for 60 min. Keratinase was concentred 4.52 times after purification and presents molar weight of 40 and 50 kDa. The purified fraction was thermal stability at 40°C for 60 min. Evaluation of chitinolytic enzymes against human lice showed an efficiency of 9.1% higher for the enzymatic extract when compared with commercial products. The evaluation of the action of keratinolytic enzyme in the degradation of human callus showed significative results when compared to products on the market less time reaching maximum degradation
Enzymes are catalysts for fundamental biological systems is extensively studied and applied worldwide, especially in health. In this study were evaluated the application of enzymes chitinolytic and keratinolytic in tests in vitro against Pedicullus humanus capitis and degradation of human callus, respectively. Were selected microorganisms with the greatest potential for production of these enzymes and determined the production parameters, conditions of separation, purification and characterization of enzymes. The microorganisms with the higher potential for the production of chitinases and keratinases were Metarhizium anisopliae CG374 and Aspergillus oryzae, respectively. The highest yields of chitinases obtained in fermentations in a bioreactor were 28°C, pH 7.0, 1.5 vvm and 200 rpm after 120 h. When produced in the hyperbaric bioreactor at a pressure of 5 bar, an increase of 21.3% in yield over conventional production in a conventional bioreactor was observed. To keratinases in a bioreactor, the production parameters were 28°C, pH 6.0, 200 rpm, and 1.5 vvm after 120h. The production of chitinases and keratinases in bioreactor was 83 and 154% higher than in Erlenmeyer flasks, respectively. The chitinase was 5.12 times concentred on the purification presenting molar weight of approximately 24, 28 and 78 kDa. Fraction containing purified chitinase showed thermal stability of 40°C for 60 min. Keratinase was concentred 4.52 times after purification and presents molar weight of 40 and 50 kDa. The purified fraction was thermal stability at 40°C for 60 min. Evaluation of chitinolytic enzymes against human lice showed an efficiency of 9.1% higher for the enzymatic extract when compared with commercial products. The evaluation of the action of keratinolytic enzyme in the degradation of human callus showed significative results when compared to products on the market less time reaching maximum degradation
Descrição
Palavras-chave
Enzimas - Produção, Quitinase, Queratinase, Enzyme, Chitinase, Keratinase
Como citar
SANTOS, Emerson dos. Produção e avaliação das aplicações de enzimas quitinolíticas e queratinolíticas. 2011. 132 f. Tese (doutorado) - Universidade Estadual Paulista, Faculdade de Ciências Farmacêuticas, 2011.