β-1,3 Glucanases e quitinases: aplicação na lise de leveduras e inibição de fungos

Abstract

RESUMOObjetivou-se, no presente trabalho, a aplicação de -1,3 glucanases e quitinases da linhagem Cellulosimicrobium cellulans 191 na lise de leveduras e inibição de fungos, respectivamente. O delineamento experimental mostrou que as melhores condições para a lise de Saccharomyces cerevisiae KL-88 pela -1,3 glucanase foi pH 6,5 e 35ºC. As células de leveduras incubadas por 10 h em frascos sem agitação mostraram-se mais susceptíveis à lise pela ação da enzima. Foi obtido maior lise da levedura quando a suspensão de células foi submetida ao tratamento com -1,3 glucanase e cisteína 1mM. A enzima invertase intracelular ou ligada à célula de S. cerevisiae KL-88 e K. marxianus NCYC 587 foi extraída após tratamento da suspensão celular com -1,3 glucanase, sendo que o tratamento prévio das leveduras com a enzima aumentou a susceptibilidade das células à lise com ultra-som. A preparação de quitinase foi capaz de formar halos de inibição de alguns fungos.Termos para indexação: -1,3 glucanases, quitinases, lise celular, inibição. ABSTRACTThe aim of this work was the application of -1,3 glucanases and chitinases by Cellulosimicrobium cellulans 191 strain on yeast cell lysis and fungi inhibition, respectively. The experimental design study showed that the best conditions to Saccharomyces cerevisiae KL-88 lysis by -1,3 glucanase extract were pH 6,5 and 35ºC. This study also demonstrated that the yeast cells were more susceptible to lysis after 10 h of cultivation in flasks without agitation. Lysis activity was increased when S. cerevisiae KL-88 cell suspension was treated with -1,3 glucanase and cystein 1mM. The enzyme invertase of S. cerevisiae KL-88 and Kluyveromyces marxianus NCYC 587 was extracted after treatment of cell suspension with -1,3 glucanase and the previous treatment of yeasts with the enzyme, increased the susceptibility to lysis when ultrasonic treatment was used. The chitinase presented growth inhibition halos for some of the fungi.

Application of Doehlert experimental design in the optimization of experimental variables for the Pseudozyma sp. (CCMB 306) and Pseudozyma sp. (CCMB 300) cell lysis

“…The organism was only able to lyse the wall in the presence of alkaline protease (SCOTT; SCHEKMAN, 1980). Fleuri and Sato (2008) produced β-1,3-glucanase from Cellulomonas cellulans 191 and used it in the analysis of Saccharomyces cerevisiae KL-88 and the extraction of intracellular enzymes. In the experimental design, these authors used temperature and pH of lysis as independent variables and used time and agitation to grow the yeasts.…”

“…Mathematical models have been increasingly used to help explain responses of biochemical reactions. This technique is often used for optimization and/or verification of the influence of medium components and cultivation parameters for enzyme production (FLEURI; SATO, 2008).…”

“…Mathematical models have been increasingly used to help explain responses of biochemical reactions. This technique is often used for optimization and/or verification of the influence of medium components and cultivation parameters for enzyme production (FLEURI; SATO, 2008).The production and optimization of the parameters that affect the enzymatic synthesis should always be investigated because the optimal conditions vary among different organisms and different enzymes (BRAVO et al, 2000). In many cases, the interaction of parameters that influence fermentation processes can be evaluated with a reduced number of tests using an experimental design (THÉODORE; PANDA, 1995).…”

Elucidação parcial da estrutura de aminoglucanooligossacarídeos (AGO's) produzidos enzimaticamente

“…A enzima (2,68 U de enzima/mL de tampão fosfato 0,01 M, pH 7,0) purificada por Fleuri [16] , foi aplicada sobre a quitina coloidal, na concentração de 6 mg do substrato/mL de enzima, em um reator tipo batelada. A reação, conduzida utilizando 10 mL da enzima sob agitação, foi mantida a 45 °C por períodos de até 1 hora.…”

beta-(1,3)-Glucanases Fúngicas: Produção e Aplicações Biotecnológicas.

“…Estas hidrolases participam diretamente do processo de controle biológico porque hidrolisam β-1,3-1,6-glucanas constituintes da parede celular de alguns patógenos (IORIO et al, 2008;FLEURI;SATO, 2008).…”