Lentinus citrinus (Walleyn e Rameloo) DPUA 1535: Crescimento, produ????o e extra????o de proteases por fermenta????o submersa

• Author(s): Kirsch, Larissa de Souza
• Date: 2015
• Origin: Oasisbr
• Subject(s): Lentinus citrinus; Proteases; Fermenta????o submersa; Lentinus citrinus; Proteases; Submerged culture; CI??NCIAS BIOL??GICAS
• Document Type: Master thesis
• Language: Portuguese

Description

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CAPES - Coordena????o de Aperfei??oamento de Pessoal de N??vel Superior

Lentinus citrinus (Walleyn e Rammeloo) DPUA 1535 from Culture Collections DPUA 1535/UFAM was analyzed as the production of proteases by submerged culture. A full factorial design 25 with three central points was established to evaluate the influence of nutrient sources, pH of the culture medium, temperature and incubation time, in the production of biomass and proteases. The extraction of proteases from culture broth with higher activity was prepared by aqueous two-phase systems, PEG/phosphate by using a full factorial design 23 with three central points. The results showed that the best condition for growth and proteases production was determinated in medium formulated with soluble starch, gelatin, pH 7,0, 25 oC in 5 days. It was found, in culture conditions, that only nitrogen source and temperature were significant in growth of L. citrinus, while for proteases production all variables were statistically significant. In PEG-phosphate systems, in all runs, the proteases moved to top phase, as well as the best conditions for extraction of enzymes were with obtained with PEG 6000, 17,5% (w/w) and phosphate 25% (w/w).

Lentinus citrinus (Walleyn e Rammeloo) DPUA 1535 da Cole????o de Culturas DPUA/UFAM foi submetido a fermenta????o submersa para a produ????o de proteases. Um planejamento fatorial completo 25 com 3 pontos centrais foi elaborado para avaliar a influ??ncia de fontes nutricionais, pH do meio de cultivo, temperatura e tempo de incuba????o na produ????o de biomassa e de proteases em cultivo submerso. A extra????o das proteases do extrato bruto de maior atividade foi realizada pelo Sistema de Duas Fases Aquosas - SDFA - PEG-fosfato, utilizando-se um planejamento fatorial 23 com 3 pontos centrais. Os resultados mostraram que a melhor condi????o para o crescimento e produ????o das proteases secretadas por L. citrinus foi determinada no meio formulado com amido sol??vel 0,5% (p/v) e gelatina 0,2% (p/v), pH 7,0, a 25 ??C, em 5 dias. Verificou-se que, nas condi????es de fermenta????o somente fonte de nitrog??nio e temperatura influenciaram no crescimento de L. citrinus, enquanto na atividade proteol??tica todas as vari??veis foram estatisticamente significativas. No processo de extra????o, em todos os ensaios do sistema PEG-Fostato, as proteases particionaram para a fase superior e as melhores condi????es de extra????o dessas enzimas foram obtidas com PEG 6000 17,5% (m/m) e Fosfato 25% (m/m).