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 O amendoim (Arachis hypogaea L.) é uma planta oleaginosa que se destaca no Nordeste Brasileiro devido a sua adaptação as condições climáticas. No entanto, existem escassas informações sobre as características das cultivares existentes na região quanto ao seu comportamento em situações de estresses e as mudanças bioquímicas das sementes. O objetivo do trabalho foi estudar as alterações fisiológicas e bioquímicas em sementes envelhecidas de amendoim. Os experimentos foram conduzidos nos Laboratórios de Análise de Sementes (LAS) e Biotecnologia do CENLAG da Universidade Federal do Agreste de Pernambuco – (UFAPE). Foram utilizadas sementes de duas cultivares: IAC Tatu ST e Caiana, envelhecidas artificialmente a 42 °C e 100 % de umidade relativa durante 96 horas, sendo retiradas amostras a cada 24 horas para avaliações fisiológicas e bioquímicas. A qualidade fisiológica e o metabolismo oxidativo foram avaliados por meio dos testes de germinação e vigor, e modificações enzimáticas (CAT e APX), respectivamente. O aumento do período de envelhecimento acelerado resultou em alterações fisiológicas e bioquímicas ocorridas nas sementes das duas cultivares, refletindo o efeito da deterioração causado pelo envelhecimento acelerado nas sementes de amendoim. A atividade das enzimas nas sementes envelhecidas de amendoim foi específica em função das cultivares, sendo que a atividade da enzima catalase foi reduzida nas sementes da cultivar Caiana, com menor vigor.

ABSTRACT Peanut (Arachis hypogaea L.) is an oleaginous plant that is well-adapted to the climatic conditions in the northeast of Brazil. However, there is little information on the existing cultivars in the region with regards to their behavior under stress conditions and the biochemical changes of the seeds. The objective of this study was to investigate the physiological and biochemical changes in aged peanut seeds. The experiments were conducted at the Seed Analysis Laboratories (LAS) and Biotechnology of CENLAG, Federal University of Agreste of Pernambuco (UFAPE). Seeds of two cultivars, IAC Tatu ST and Caiana, were artificially aged at 42 °C and 100 % relative humidity for 96 h, and removed every 24 h for evaluation. Physiological potential was evaluated using germination, vigor test, and enzymatic modifications as the parameters. An increase in the accelerated aging period resulted in physiological and biochemical changes in the seeds of both cultivars. The activity of the enzymes in the aged peanut seeds was specific to the cultivar used, and the seeds of the cultivar Caiana had less vigor and decreased activity of the enzyme catalase.

RESUMO A Caatinga, bioma exclusivamente brasileiro vem sofrendo elevada ação antrópica, o que interfere diretamente na qualidade de seu solo. Para avaliar e acompanhar a qualidade do solo vem-se utilizando indicadores químicos e biológicos, com a determinação da atividade enzimática, ganhando destaque. Neste sentido, objetivou-se avaliar a atividade enzimática de solos do bioma Caatinga em áreas preservadas e antropizadas, pertencentes ao Parque Nacional do Catimbau, situado em Pernambuco, Brasil, nos dois períodos do ano (chuvoso e seco), por três anos consecutivos, 2014, 2015 e 2016. Foram avaliadas as atividades enzimáticas de β-glicosidase, urease, arilsulfatase, fosfatase ácida e alcalina de solos. A ação antrópica proporcionou redução das atividades enzimáticas de β-glicosidase, urease e arilsulfatase. Entre os períodos do ano, o seco apresentou menores valores de atividade enzimática de β-glicosidase, urease e arilsulfatase. Entre os anos de avaliação, ao correlacionar a precipitação acumulada e a atividade enzimática, as maiores correlações foram observadas na área preservada em ambos períodos do ano. Comprovou-se a fragilidade dos solos da Caatinga sob a ação antrópica em ambas as estações dos períodos do ano, ao longo do tempo. A antropização crescente de áreas do Parque Nacional do Catimbau apresenta efeito direto na qualidade do solo, afeta seu equilíbrio ecológico, causando a sua degradação e redução de sua qualidade. Tornam-se necessárias medidas urgentes de fiscalização adequada para garantir a sua preservação ambiental.

ABSTRACT The Caatinga is an exclusively Brazilian biome that has been under high anthropogenic action, which directly interferes in the quality of its soil. In order to evaluate and monitor the quality of the soil, chemical and biological indicators are used, with the determination of the enzymatic activity gaining prominence. The objective of this study was to evaluate the enzymatic activity of soils of the Caatinga biome in preserved and anthropized areas belonging to the Catimbau National Park, located in Pernambuco, Brazil, during the two periods of the year (rainy and dry), for three consecutive years, 2014, 2015 and 2016. Enzymatic activities of β-glucosidase, urease, arylsulfatase, and acid and alkaline phosphatase in the soil were evaluated. Anthropic action caused reduction in the enzymatic activities of β-glucosidase, arylsulfatase and urease. Regarding the periods of the year, the dry one showed smaller values of enzymatic activity of β-glucosidase, arylsulfatase and urease. Among the evaluation years, when correlating accumulated precipitation and enzymatic activity, the highest correlations were observed in the preserved area in both periods of the year. The fragility of Caatinga soils under anthropic action was observed in both periods of the year, over time. The increasing anthropization of areas of the Catimbau National Park has a direct effect on soil quality, affects its ecological balance, causing degradation and reduction of its quality. Appropriate enforcement measures are urgently needed to ensure their environmental preservation.

RESUMO O feijão caupi, [Vigna unguiculata (L.) Walp.] tem uma grande importância em virtude do elevado teor proteico e energético na alimentação humana. Dentre as principais doenças da cultura do feijão, destaca-se a antracnose, causada pelo desenvolvimento do fungo hpsColletotrichum lindemuthianumhps. A indução de resistência torna-se uma alternativa para controle desse micro-organismo, entre os indutores mais utilizados, destaca-se o acibenzolar-S-metil (ASM). Dessa forma, o presente trabalho teve como objetivo avaliar a eficiência do ASM em feijão caupi no controle da antracnose, causada pelo fungo hpsColletotrichum lindemuthianumhps URM 5771, bem como a ativação de enzimas relacionadas à patogenicidade. O estudo foi conduzido em casa de vegetação e os tratamentos foram dispostos em um esquema de delineamento inteiramente casualizado, sendo cinco tratamentos (Tratamento 1 - 0,15g L-1; Tratamento 2 - 0,30g L-1; Tratamento 3 - 0,45g L-1 e Tratamento 4 - 0,60g L-1), incluindo o controle (apenas água) e quatro repetições. Os tratamentos com maiores doses do ASM proporcionaram maior atividade enzimática e consequentemente maior resistência às plantas de feijão caupi.

ABSTRACT Cowpea [Vigna unguiculata (L.) Walp.] is of great importance due to its high protein and energy content for human diet. Among the major diseases of bean culture is anthracnose, caused by the development of the fungus Colletotrichum lindemuthianum. Resistance induction becomes an alternative to control this microorganism; among the most commonly used inducers is acibenzolar-S-methyl (ASM). Thus, the present study aimed to evaluate ASM efficiency in controlling anthracnose in cowpea, which is caused by the fungus Colletotrichum lindemuthianum URM 5771, as well as the activation of enzymes related to its pathogenicity. The study was conducted in a greenhouse and treatments were arranged in a completely randomized design, constituting 5 treatments (Treatment 1 - 0.15g L-1; Treatment 2 - 0.30g L-1; Treatment 3 - 0.45g L-1 and Treatment 4 - 0.60g L-1), including control (water only) and four replicates. Treatments with higher ASM levels provided higher enzymatic activity and, consequently, greater resistance to cowpea plants.

Abstract Protein hydrolysates originating from egg white have already been reported to be bioactive and, among their biological activities, possess the antioxidant property that protects the body from early ageing and diseases linked to oxidation. Therefore, the objective of this work was to evaluate the antioxidant activity of hydrolysates obtained by the hydrolysis of egg white from hen poultry. The protease produced by Aspergillus avenaceus URM 6706 was purified and subsequently applied to hydrolysate the egg white, and the degree of hydrolysis was verified during the protease exposure time (4-24 h). The hydrolysis was intensified over time of exposure to the protease. It was possible to detect the antioxidant activities of eliminating the 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) radical (ABTS•+) from 97% to 99% and 2,2-diphenyl-1-picrylhydrazyl (DPPH•) up to 27%, as well as the chelation of Cu2+ metal ions up to 62% and Fe2+ up to 54%. The elimination of ABTS•+ radical had a positive correlation with the degree of hydrolysis; however, all the other activities tested showed a negative correlation with the degree of hydrolysis. The results obtained suggest that the egg white of hen chicken represents a food source of animal origin with potential application in the functional food industry.

Abstract Different responses are observed in plants after infection by pathogens. The usage of systemic inducers, whether biotic or abiotic, has the function to increase the enzymatic production of the plant defense system. In that sense, our present research aims to evaluate the disease severity of anthracnose in the cowpea cultivar “sempre-verde” infected with Colletotrichum lindemuthianum URM 5771 and subjected to both biotic and abiotic resistance inducers, as well as evaluate the enzymatic activity of plant defense. The experiment was performed in a greenhouse with a completely randomized split-plot design with six treatments: four abiotic plant resistance inducers, acibenzolar-S-methyl, β-aminobutyric acid, chitosan, and salicylic acid; one biotic inducer, the filamentous fungi Trichoderma aureoviride URM 5158; and a control with only water. The plant leaves were sampled at four time points, 4, 8, 12 and 20 days after inoculation with the plant pathogen, to measure the enzymatic activity of β-1,3-glucanase, catalase, peroxidase, polyphenoloxidase, and protease. The abiotic inducers acibenzolar-S-methyl, β-aminobutyric acid, and chitosan and the biotic inducer presented the highest potential in reducing the anthracnose caused by C. lindemuthianum URM 5771 in cowpea plants by showing the lowest values of injury severity over time. The disease progression showed a proportionate shift in enzymatic activity in that the inducers salicylic acid and T. aureoviride URM 5158 promoted the highest enzymatic activity when compared to the other inducers. High correlations between the enzymatic activity of peroxidase and polyphenoloxidase and anthracnose severity were observed in cowpea plants. The use of abiotic and biotic resistance inducers presents high potential for anthracnose control in cowpea plants; however, more studies should be carried out covering different environments, plant cultivars and climatic conditions.

Resumen Se observan diferentes respuestas en la planta después de la infección por patógenos. El uso de inductores sistémicos ya sean bióticos o abióticos tiene la función de aumentar la producción enzimática del sistema de defensa de la planta. En ese sentido, nuestra presente investigación tiene como objetivo evaluar la severidad de la enfermedad en plantas de caupí cultivares “sempre-verde” infectadas con Colletotrichum lindemuthianum URM 5771, el agente de la antracnosis, y sometidas a inductores de resistencia tanto bióticos como abióticos, así como estimular la enzimática actividad de defensa de planta. El experimento se realizó en invernadero con un diseño de parcelas divididas completamente aleatorizado en el tiempo, con seis tratamientos: cuatro inductores de resistencia de plantas abióticas acibenzolar-S-metilo, ácido β-aminobutírico, quitosano y ácido salicílico, un inductor biótico, los hongos filamentosos Trichoderma aureoviride URM 5158, y un control con solo agua. Las hojas de la planta se muestrearon en cuatro puntos de tiempo, 4, 8, 12 y 20 días después de la inoculación del patógeno de la planta, para medir la actividad enzimática de la β-1,3-glucanasa, la catalasa, la peroxidasa, la polifenoloxidasa y la proteasa. Los inductores abióticos acibenzolar-S-metil, β-aminobutírico, quitosano y el inductor biótico presentaron el mayor potencial para reducir la antracnosis causada por C. lindemuthianum URM 5771 en las plantas de caupí al mostrar los valores más bajos en la escala de severidad de las lesiones a lo largo del tiempo. La progresión de la enfermedad proporcionó un cambio en la actividad enzimática donde los inductores de ácido salicílico y T. aureoviride URM 5158 promovieron la actividad enzimática más alta en comparación con los otros inductores. Se observaron altas correlaciones entre la actividad enzimática de la peroxidasa y la polifenoloxidasa y la severidad de la antracnosis en las plantas de caupí. El uso de inductores de resistencia abióticos y bióticos presenta un alto potencial para la lucha contra la antracnosis en las plantas de caupí, sin embargo, se deben realizar más estudios que cubran diferentes ambientes, variedades de plantas y condiciones climáticas.

ABSTRACT The Caatinga biome features an exclusive endemic biodiversity, and is characterized by the presence of xerophytic, deciduous vegetation, high temperatures, and low rainfall. This important park has undergone anthropization, especially through extraction of firewood and timber and growing plants for raising goats. The objectives of this study were to compare the communities of filamentous fungi present in the preserved area and in the anthropized soil of the Catimbau National Park in Buíque, PE, Brazil, and to evaluate the impacts of anthropization on such communities. A total of 12 collections of soil samples were made, six in the preserved area and six in the anthropic area, and the physicochemical properties of the soil samples were analyzed. Fungi were isolated through suspension and serial dilution methods. After growth, the samples were purified and identified based on classical taxonomy, according to specific literature. The diversity, evenness, richness, dominance, frequency, and similarity among the species of filamentous fungi in both areas were assessed based on ecological indexes. A total of 4,488 colony-forming units of filamentous fungi were obtained, which were distributed into 65 species belonging to 15 genera. In the preserved area, higher abundance and richness of species were observed, with predominance of the genera Aspergillus and Penicillium. In both areas, diversity and equitability were high, demonstrating that the species are well distributed in these areas. In the preserved area, the dominant genera were Aspergillus, Gongronella, and Penicillium, whereas Aspergillus was the dominant genus in the anthropic area. Two distinct communities were observed in the areas analyzed. Principal component analysis showed that Penicillium simplicissimum influences the total diversity of both communities. The anthropization that occurred in the Catimbau National Park has changed the composition of the filamentous fungal communities of the site, restricting the number of species and decreasing the abundance of these important microorganisms. This results in ecosystem damage and likely causes relevant major imbalances, with serious consequences, such as possible disappearance of the aforementioned species, as well as of species yet undiscovered by the scientific community.

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

A mandioca é utilizada na alimentação humana, animal e na indústria. No Estado da Paraíba, é explorada comercialmente no Litoral, na microrregião do Brejo Paraibano e como cultura de subsistência no interior. Tendo em vista a ausência de estudos epidemiológicos da cultura no Estado, objetivou-se com este trabalho avaliar o progresso da severidade e da incidência da mancha parda (Cercosporidium henningsii), mancha branca (Phaeoramularia manihotis) e antracnose (Colletotrichum gloeosporioides) em condições de infecção natural em campo ao longo de um ano, no Sítio Geraldo, situado no município de Alagoa Nova-PB, Brejo paraibano. Os dados para os estudos epidemiológicos foram registrados no 18º dia de cada mês, no período de setembro de 2012 a agosto de 2013. Para avaliar a incidência, calculou-se a porcentagem de plantas apresentando sintomas. A severidade foi quantificada a partir das observações das lesões nos tecidos foliares com o emprego de escala diagramática, escala descritiva e índice de Índice de Mackinney. Os dados da curva de progresso foram submetidos à análise de correlação de Pearson em relação aos dados meteorológicos mensais. Calculou-se também a área abaixo da curva de progresso da doença (AACPD). Dentre as plantas selecionadas 95%, 17% e 8,5% apresentaram os sintomas de mancha parda, mancha branca e antracnose, respectivamente. As lesões destas doenças foram observadas na ordem de 13,3%; 4,7% e 3,3% de extensão nos tecidos aéreos. Os valores de incidência e severidade das três doenças aumentaram gradativamente no decorrer do período de avaliação, atingindo o resultado final de 61,02; 32,45 e 15,28 para AACPD. As taxas de progresso da severidade mensais mais elevadas foram de 0,441 para mancha parda em setembro, 0,197 para mancha branca e 0,192 para antracnose, em novembro e dezembro. Houve correlação positiva entre o índice pluviométrico e o progresso das doenças e negativa em relação à diminuição das temperaturas médias mensais para as três enfermidades.

The cassava is used in human and animal food, as well as in the industry. In the state of Paraíba, it is commercially explored on the Coast, in the microregion of "Brejo Paraibano" and as subsistence crop in inner regions of the state. In view of the lack of epidemiological studies of this crop in Paraiba State, the aim of this study was to evaluate the severity progress and the incidence of brown leaf spot (Cercosporidium henningsii), anthracnose (Colletotrichum gloeosporioides) and white leaf spot (Phaeoramularia manihotis) under conditions of natural infection in the field over one year at the small farm Geraldo, in the city of Alagoa Nova-PB, "Brejo Paraibano". The data used for the epidemiological studies were recorded on the 18th day of every month, from September 2012 t oAugust 2013. Toassess the incidence, the percentage of plant sex hibiting symptoms was calculated. Disease severity was quantified based on observations of lesions in leaf tissues by using a diagrammatic scale, descriptive scale and the Mackinneyindex. The disease progresscurve data were subjected to Pearson correlation analysis relative to the monthly meteorological data. The area under the disease progresscurve (AUDPC) was also calculated. Of the selected plants, 95%, 17% and 8.5% showed symptoms of brown leaf spot, white leaf spot and anthracnose, respectively. The lesions of these diseases were observed in the order of 13.3%, 4.7% and 3.3% of extension in shoot tissues. The values of incidence and severity of the three diseases gradually increased over the evaluation period, reaching the final results of 61.02, 32.45 and 15.28 for AUDPC. The highest monthlyrates of severity progress were 0.441 for brown leaf spotin September, 0.197 for white leaf spotand 0.192 for anthracnose in November and December. There was a positive correlation between the pluviometric indexand the disease progressand a negative correlation for the decline in the monthly average temperatures for the three diseases.

Objetivando-se levantar e quantificar a incidência de doenças da mandioca em plantios comerciais no Estado da Paraíba, realizaram-se coletas em 21 municípios de quatro microrregiões do Estado no primeiro semestre de 2011. As doenças detectadas foram mancha parda (Cercosporidium henningsii), mancha branca (Phaeoramularia manihotis), antracnose (Colletotrichum gloeosporioides f. sp. manihotis) e podridão mole (Fusarium oxysporum). A incidência apresentou-se com variação, sendo os maiores índices constatados nos municípios das microrregiões do Curimataú Ocidental, Sapé e Brejo paraibano. A mancha parda ocorreu em todos os municípios avaliados, sendo seguida da mancha branca, cuja porcentagem de plantas atacadas foi inferior. A antracnose foi detectada em cerca de 70% dos municípios avaliados, porém com incidência reduzida. A maior incidência da podridão causada por F. oxysporum foi observada na cidade de Imaculada, microrregião de Serra do Teixeira.

To survey and quantify the incidence of the diseases affecting cassava (Manihot esculenta) in commercial crops of the state of Paraiba - Brazil, samplings were conducted in 21 municipalities at four microregions of the state during the first semester of 2011. The detected diseases were brown leaf spot (Cercosporidium henningsii), white leaf spot (Phaeoramularia manihotis), anthracnose (Colletotrichum gloeosporioides f. sp. manihotis) and root rot (Fusarium oxysporum). The incidence varied, and the highest indexes were found for the municipalities of the microregions of Curimataú Ocidental, Sapé and Brejo Paraibano. Brown leaf spot occurred in all evaluated municipalities, followed by white leaf spot, which showed inferior percentage of affected plants. Anthracnose was detected for around 70% of the evaluated municipalities but had reduced incidence. The highest incidence of root rot caused by F. oxysporum was observed for the city Imaculada, microregion of the Serra do Teixeira.

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.

Enzymes obtained by fermentation processes offer a number of advantages and have been widely researched and used throughout the world. This study aimed to partially characterise an inulinase produced from palm and cassava peel. The enzyme was produced via the solid-state fermentation of Aspergillus japonicus URM5633. The optimal temperatures were 50ºC and 55ºC, and the optimal pH values were 5.2 and 3.4 for inulinase fermentatively produced from palm and cassava peel, respectively. The thermostability measurements for inulinase produced in palm showed that the relative activity remained below 100% until 30 minutes of stability for all temperatures, but reached 106.8% at a temperature of 50ºC after 60 minutes. Inulinase from the crude extract of cassava peel was pH stable and only decreased to 55% of the maximal activity over the course of the assay, suggesting that this enzyme can be used in inulinase production and can be utilized in food industries.

A enzima coagulante de leite obtida de látex de raiz de Jacaratia corumbensis O. kuntze foi caracterizada parcialmente e purificada, por precipitação fracionária com sulfato de amônio e cromatografia de troca de íon. Foram utilizadas cinco frações de sulfato de amônio (AS1 - 0-20%; AS2 - 20-40%; AS3 - 40-60%; AS4 - 60-80%; AS5 - 80-100%), a fração 40-60% (AS3) mostrou alta atividade coagulante com um fator de purificação de 1,2 vezes em relação ao extrato bruto. Esta fração foi aplicada em coluna Mono Q obtendo dois picos de proteína (p1 e p2), o p1 mostrou melhor atividade coagulante. O pH ótimo para o extrato bruto e parcialmente purificado foi 6,5 e 7,0, respectivamente. A atividade coagulante foi atingida a 55ºC para ambos os extratos, bruto e parcialmente purificado. A enzima foi inibida por ácido iodoacético que sugere que esta enzima é uma cisteína protease, com peso molecular de 33 kDa.

The partial characterization and purification of milk clotting enzyme obtained from the (root latex) of Jacaratia corumbensis O. kuntze was studied, by fractional precipitation with ammonium sulphate and ion exchange chromatography. The ammonium sulphate precipitate showed five fractions (AS1- 0-20%; AS2 - 20-40%; AS3 - 40-60%; AS4 - 60-80%; AS5 - 80-100%) and among the fractions obtained, the 40-60% fraction (AS3) showed the highest milk clotting activity with a purification factor of 1.2 fold in relation to the crude extract. This fraction when applied on Mono Q column yielded two protein peaks (p1 and p2), but p1 pool showed the best milk-clotting activity. The optimal pH for the crude and partially purified extract was 6.5 and 7.0, respectively. The maximum milk-clotting activity was at 55ºC for the both crude and partially purified extracts. The enzyme was inhibited by iodoacetic acid which suggested that this enzyme was a cysteine protease, with molecular weight of 33 kDa.

Enzimas amilolíticas têm sido amplamente investigadas com a finalidade de melhorar os processos industriais para a degradação do amido. Foi determinado que a extração da enzima em sistema bifásico aquosos é um método aplicável para separação e purificação de biomoléculas em misturas. Vários sistemas compostos de soluções aquosas de polietilenoglicol e fosfato foram avaliados. Estudos de produção em meio mínimo suplementado, à 37ºC, com uma velocidade de agitação de 110rpm e recuperação da amilase a partir do Bacillus subtilis em sistema bifásico aquoso PEG-fosfato foram avaliados. O melhor fator de purificação (5.4) foi obtido no sistema PEG 1000 (16.7% w/w) com fosfato de potássio (14.8% w/w), a pH 6.0, resultando na recuperação da atividade enzimática de 45.2% na fase rica em sal.

Studies were carried out on the partition of amylase from Bacillus subtilis in a minimal medium at 37 ºC and 110 rpm. Enzyme recovery was carried out in aqueous two-phase system PEG-Phosphate salt were carried out. The best purification factor (5.4) was obtained in system PEG 1000 (16.7% w/w) with potassium phosphate (14.8% w/w), at pH 6.0, resulting in a recovery of 45.2% activity enzymatic in the salt-rich phase.