RESUMO Objetivou-se com este trabalho otimizar a produção de enzimas celulolíticas por hps C. gloeosporioideshps URM 7080 isolado de lesões foliares de Manihot esculenta, sintomáticas de Antracnose. Utilizou-se farelo de entrecasca de mandioca (MA) e palma forrageira triturada (PA) como substratos para fermentação submersa nas concentrações 0,5, 0,75 e 1%, pH inicial 5,0, 6,0 e 7,0; agitações 80, 110 e 140 rpm e temperaturas de 28, 32 e 36 °C em experimento fatorial fracionário 25 com 32 ensaios + 4 repetições no ponto central. Os melhores parâmetros para produção de cada enzima isolada foram: 1% de MA, 0,5% de PA, 28° C, com o pH inicial 5,0 para FPAse (Celulase total) e pH 7,0 para CMCase (Endoglucanase); Para βG (β-glucosidase), pH inicial 5,0 à 36° C, e concentração de 0,5 de PA e agitação de 140 rpm para as três enzimas em conjunto.

ABSTRACT The aim of the present study was to optimize the production of cellulolytic enzymes by C. gloeosporioides URM 7080 isolated from leaf lesions in Manihot esculenta showing symptoms of anthracnose. Cassava bran (MA) and crushed forage cactus (PA) were used as substrates for submerged fermentation at the concentrations of 0.5%, 0.75% and 1%; initial pH of 5.0, 6.0 and 7.0; agitation at 80, 110 and 140 rpm, and temperatures of 28, 32 and 36°C, in a 25 fractional factorial experiment with 32 trials + 4 replicates at the central point. The best parameters for the production of each isolated enzyme were: 1% MA, 0.5% PA, 28°C, and initial pH of 5.0 for FPase (total cellulase) and pH of 7.0 for CMCase (endoglucanase); initial pH of 5.0 at 36°C for βG (β-glucosidase), and concentration of 0.5% PA at 140 rpm for the three enzymes together.

RESUMO A partir de coletas realizadas em plantios comerciais de mandioca (Manihot esculenta Crantz) em 11 municípios do Estado da Paraíba, obtiveram-se 27 isolados dos fungos Colletotrichum gloeosporoides f. sp. manihotis Henn (Penn.) e Fusarium oxysporum Schlent, fitopatogênicos à cultura. Desses isolados foi obtida a produção enzimática do complexo celulolítico, sob fermentação submersa à 120 rpm e 30 °C durante 7 dias, tendo CMC (Carboximetilcelulose) como fonte de carbono. As atividades enzimáticas foram quantificadas por meio do grau de hidrólise de tiras de papel de filtro (FPAse) e de CMC (CMCase). Utilizou-se a liberação de Paranitrofenol para quantificação das atividades da β-glucosidase (βG). As atividades mais elevadas observadas foram: 23,13 e 1,51 U/ml para FPAse e CMCase pelos isolados de C. gloeosporioides URM 7124 e 7125. Para βG, 0,0014 U/mL, pelo isolado 13.

ABSTRACT Material collected from commercial crops of cassava (Manihot esculenta Crantz) in 11 municipalities in the state of Paraíba included 27 isolates of the fungi Colletotrichum gloeosporoides f. sp. manihotis Henn (Penn.) and Fusarium oxysporum Schlent., which are pathogenic to the culture. From these isolates, cellulolytic enzyme complex production was obtained under submerged fermentation at 120 rpm and 30 °C for 7 days, with CMC (Carboxymethyl Cellulose) as carbon source. Enzyme activities were quantified by means of the degree of hydrolysis of strips of filter paper (FPAse) and CMC (CMCase). The release of Paranitrofenol was used to quantify the activities of β-glucosidase (βG). The highest activities were: 23.13 and 1.51 U/ml for FPase and CMCase by the isolates of C. gloeosporioides URM 7124 and 7125. For βG, 0.0014 U/ml by isolate 13.

Breadfruit is an exotic tree in Brazil, very well acclimatized. Despite of being a laticifer plant, the knowledge about its latex is scarce. However, it is known that proteolytic enzymes represents over 50% of the latex composition in laticifers plants. The aim of this study was to evaluate the potential of breadfruit (Artocarpus altilis var. Apyrena) latex as a source of milk clotting proteases and partially characterize it. The proteolytic activity of the fraction of crude extract was assessed using azocasein and quantitation of total protein, the bicinchoninic acid (BCA). Milk-clotting activity was analyzed using skim milk at 12%. The enzyme activity was analyzed under different temperature conditions (35 to 80°C) and pH (5.8 to 10.7) presenting optimum activity in alkaline pH (8.5) and 50°C; being stable to the two variables at 120 minutes during the test. The clotting activity was directly proportional to the temperature at the better concentration of CaCl2 (10mmol L-1). The results indicate that enzyme is a possible replacement for calf rennet

There are several obstacles to the use of chymosin in cheese production. Consequently, plant proteases have been studied as possible rennet substitutes, but most of these enzymes are unsuitable for the manufacture of cheese. The aim of this study was to evaluate the potential of latex from Sideroxylon obtusifolium as a source of milk-clotting proteases and to partially characterize the enzyme. The enzyme extract showed high protease and coagulant activities, with an optimal pH of 8.0 and temperature of 55 °C. The enzyme was stable in wide ranges of temperature and pH. Its activity was not affected by any metal ions tested; but was inhibited by phenylmethanesulfonyl fluoride and pepstatin. For the coagulant activity, the optimal concentration of CaCl2 was 10 µmol L- 1. Polyacrylamide gel electrophoresis showed four bands, with molecular weights between 17 and 64 kDa. These results indicate that the enzyme can be applied to the cheese industry.