OBTENÇÃO E UTILIZAÇÃO DA ENZIMA POLIFENOLOXIDASE EXTRAÍDA DE POLPA DE PINHA (Annona squamosa L.) MADURA NO MELHORAMENTO DO SABOR DO CACAU (Theobroma cacao L.)

O presente trabalho teve como objetivo estudar a obtenção e a utilização da enzima polifenoloxidase (PPO) extraída de polpa de pinha madura na redução do teor de compostos polifenólicos com a finalidade de diminuir a adstringência e o amargor das amêndoas de cacau processadas na forma de "nibs". A PPO foi extraída com tampão fosfato de potássio 0,025M (pH 7,5), adicionando sulfato de amônio para a precipitação da enzima. O material em pó obtido foi denominado de enzima parcialmente purificada, sendo que a análise de atividade enzimática foi realizada, utilizando-se de catecol como substrato. As características bioquímicas apresentadas foram pH de estabilidade de 6,0 a 6,5 e temperatura de estabilidade de 10 a 30°C. Os "nibs" foram autoclavados (121°C por 15 minutos) e não autoclavados de amêndoas cruas insuficientemente fermentadas e secas, da mesma origem, sendo embebidas em 25 mL de uma solução da enzima contendo 200 unidades/min/mL, durante 30; 60; 90; 210 e 360 minutos, a 23°C e pH 6,0. Os "nibs" foram homogeneizados com a solução de enzima a cada 15 minutos, secos, moídos e desengordurados. Após o tratamento enzimático durante 210 minutos realizado nos "nibs" de cacau desengordurado não autoclavados foi possível observar diminuição de 15% nas concentrações de fenóis totais, 15% de taninos, 10% de flavan-3-ois e 18% de antocianidinas. Os "nibs" de cacau desengordurado autoclavados apresentaram diminuição de 25% nas concentrações de fenóis totais, 26% de taninos, 23% de flavan-3-ois e 51% de antocianidinas.

Efeitos do alagamento do substrato no crescimento e na composição química de genótipos clonais de Theobroma cacao L.

O alagamento do solo pode promover alterações no metabolismo celular e causar desvios nas condições ótimas de crescimento das plantas, gerando uma condição de estresse. Objetivou-se no presente trabalho avaliar os efeitos do alagamento no crescimento e na nutrição mineral de seis clones de T. cacao (CP-49, CCN-10, CP-06, CEPEC-2007, CEPEC-2008 e PS-1319), para elucidar possíveis mecanismos de tolerância ao alagamento. Mudas clonais de T. cacao, com 6 meses de idade, foram submetidas ao alagamento, juntamente com o tratamento-controle (não alagado), por 30 dias. Observou-se, no final desse período, que o alagamento promoveu diminuição nas taxas de crescimento relativo radicular (exceto para CP-06) e de área foliar, acúmulo de matéria seca e incremento nas taxas de crescimento relativo caulinar (exceto para os clones CP-06 e CEPEC-2008) e assimilatória l��quida (exceto para o clone alagado PS-1319) e de massa foliar específica para os clones CP-49, CCN-10 e CP-06. Os clones que sobreviveram aos 30 dias de alagamento apresentaram baixos valores de razão de área foliar. De modo geral, o alagamento do substrato acarretou deficiência na absorção de macro e micronutrientes minerais, exceto de Fe. Dentre os clones avaliados, o CP-49 foi tolerante e o CEPEC-2008 não tolerante ao alagamento. Os demais clones ficaram numa posição intermediária. Logo, o clone CP-49 tem grande potencial para ser cultivado em condições de alagamento.

Aproveitamento tecnol��gico do "mel de cacau" (Theobroma cacao L) na produção de geleia sem adição de açúcar

O "mel de cacau" é uma denominação regional do l��quido transparente extraído da polpa do cacau, que cobre as sementes, antes do processo de fermentação do cacau. Este material é atrativo do ponto de vista de seus aspectos sensorial e tecnol��gico; entretanto, seu uso para o consumo ainda é restrito. O propósito deste estudo foi produzir geleia dietética usando o "mel de cacau" e realizar a caracterização química e sensorial dos produtos. Para a preparação das geleias, foram usados os seguintes ingredientes: pectina de baixo grau de metoxilação, polidextrose, sorbitol, maltitol, fosfato tricálcico, sorbato de potássio e cacau em pó. As geleias dietéticas estudadas foram: F1 (0,0015% taumatina e 0,005% sucralose); F2 (0,0015% taumatina e 0,002% acessulfame-k); F3 (0,005% sucralose e 0,002% acessulfame-k). Foi produzido, também, um tratamento-controle (com 40,0% de sacarose). Os dados químicos e sensoriais foram analisados por ANOVA, teste de Tukey e Mapa de Preferência Interno (MDPREF). O tratamento F3 (sucralose e acessulfame-k) foi a geleia dietética preferida dos consumidores. Esta diferiu significativamente (p<0.05) dos tratamentos F1 (taumatina e sucralose) e F2 (taumatina e acessulfame-k) em relação aos atributos cor, aroma de chocolate, gosto doce, gosto ácido, sabor de chocolate e consistência; obteve, também, maior nota para intenção de compra. O uso do adoçante taumatina mostrou-se inadequado por não prover doçura à geleia de "mel de cacau". Os resultados indicaram a viabilidade do aproveitamento tecnol��gico do "mel de cacau" como matéria-prima na preparação de geleia dietética, que apresentou boas características sensoriais e nutricionais.

Performance pol��nica em cacaueiros (Theobroma cacao L.) autocompatíveis e autoincompatíveis

A importância econômica do cacaueiro se deve, principalmente, à produção de frutos, dos quais se extraem sementes que servem como matéria prima para a fabricação do chocolate. Vários fatores podem interferir na produção, porém as características reprodutivas do cacaueiro e sua influência na frutificação não têm sido intensamente estudadas. Al��m disso, alguns genótipos são autoincompatíveis, mas clones autocompatíveis são encontrados em populações comerciais. Com o objetivo de comparar a performance do pólen e o modo de reprodução entre clones autocompatíveis CCN51 e autoincompatíveis TSH1188, foram realizadas estudos de germinação in vitro (utilizando-se dois testes) e da razão pólen-óvulo (P:O). O Teste 1 (meio de cultura sem alguns elementos essenciais e com maior concentração de sacarose) proporcionou melhores resultados de germinação do pólen, com médias acima de 77%, enquanto que com o Teste 2 o máximo de pólen germinado foi, em média, de 39,95%. O percentual de pólen germinado in vitro foi maior em plantas autoincompatíveis. Com base na razão P:O, ambos os clones foram classificados como autógamos obrigatórios, o que foi considerado um desvio da classificação de Cruden.

Multivariate genetic divergence and hybrid performance of cacao (Theobroma cacao L.)

Genetic distances among cacao cultivars were calculated through multivariate analysis, using the D2 statistic, to examine racial group classification and to assess heterotic hybrids. A 5 x 5 complete diallel was evaluated. Over a five-year period (1986-1990), five cultivars of the S1 generation, pertaining to the Lower Amazon Forastero and Trinitario racial groups and 20 crosses between the corresponding S0 parents were analyzed, based upon five yield components - number of healthy and collected fruits per plant (NHFP and NCFP), wet seed weight per plant and per fruit (WSWP and WSWF), and percentage of diseased fruits per plant (PDFP). The diversity analysis suggested a close relationship between the Trinitario and Lower Amazon Forastero groups. A correlation coefficient (r) was calculated to determine the association between genetic diversity and heterosis. Genetic distance of parents by D2 was found to be linearly related to average performance of hybrids for WSWP and WSWF (r = 0.68, P &lt; 0.05 and r = 0.76, P &lt; 0.05, respectively). The heterotic performance for the same components was also correlated with D2, both with r = 0.66 (P &lt; 0.05). A relationship between genetic divergence and combining ability effects was suggested because the most divergent cultivar exhibited a high general combining ability, generating the best performing hybrids. Results indicated that genetic diversity estimates can be useful in selecting parents for crosses and in assessing relationships among cacao racial groups.

Perfil sensorial de pó de cacau (Theobroma cacao L.) alcalinizado

A Análise Descritiva Quantitativa(ADQ) foi empregada para caracterização das amostras de pós de cacau que representaram a amplitude do delineamento composto rotacional central 2 do processo de alcalinização dos "nibs" de cacau. As variáveis independentes foram faixas de temperatura de 60 a 120 ºC, de tempo de 30 a 150min e de concentração de K2CO3 de 1,22 a 4,78%. Foram avaliadas oito amostras de pó de cacau representativas das variações de cor e aceitabilidade do total das amostras obtidas experimentalmente e duas amostras de marcas comerciais. A análise do aroma foi feita diretamente nos pós de cacau alcalinizados e a dos demais atributos na forma de bebida achocolatada (2% do pó de cacau e 7% de açúcar em leite desnatado esterilizado). Doze provadores selecionados com base no seu poder de discriminação, reprodutibilidade e concordância com a equipe geraram em consenso, pelo método de rede (Kelly's Repertory Grid Method), três termos descritos para o aroma (alcalino, chocolate e queimado) e doze para os achocolatados (solubilidade, marrom, marrom avermelhado, chocolate, queimado, caramelo, doce, adstringente, alcalino, amargo, salgado e corpo). As avaliações das amostras foram feitas monadicamente com três repetições e em cabines individuais. Os dados obtidos foram submetidos a ANOVA, teste de Tukey e Análise de Componentes Principais. As avaliações dos aromas dos pós de cacau mostraram relação direta entre o aroma alcalino e os teores de álcali, temperatura e tempo do processo. De modo geral, os produtos com menores concentrações de K2CO3 (1,22 - 3,00% ) apresentaram aroma e sabor de chocolate mais fortes. Encontrou-se uma relação direta entre os teores de álcali no produto e o sabor alcalino, queimado e adstringente e uma relação inversa com a luminosidade da cor. Assim, as amostras com maior concentração de K2CO3 (4,78%), foram consideradas pela equipe sensorial, as de mais forte sabor e aroma alcalino, queimado e adstringente, assim como de cor marrom e marrom avermelhado mais escura. Todos os processos levaram à obtenção de amostras com alta solubilidade e fraco sabor amargo.

Nutritional value and antioxidant capacity of "cocoa honey" (Theobroma cacao L.)

Cocoa honey is considered as the liquid portion of cocoa pulp that is released from the fruit soon after it is cut open and can be used before fermentation by simple extraction due to its nutritional characteristics. The objective of the present study is to determine the biochemical characteristics of a cocoa by-product, "cocoa honey" (CH), produced in the State of Bahia-Brazil. The biochemical characterization was conducted to determine reducing sugars, total sugars, vitamin C, total dietary fiber, flavonoids, and total antioxidant activity using an EC50. It was observed that cocoa honey can be considered a source of bioactive compounds, can be consumed in natura or processed, and used as an ingredient in the chocolate industry and in other food products. However, it is necessary to use complementary methods, such as HPLC, to quantify the phenolic compounds of this by-product.

Bioactive compounds in different cocoa (Theobroma cacao, L) cultivars during fermentation

One component that contribute to the flavor and aroma of chocolate are the polyphenols, which have received much attention due to their beneficial implications to human health. Besides bioactive action, polyphenols and methylxantines are responsible for astringency and bitterness in cocoa beans. Another important point is its drastic reduction during cocoa processing for chocolate production and the difference between cultivars. Thus, the present study aimed to evaluate the modifications in monomeric phenolic compounds and methylxanthines during fermentation of three cocoa cultivars grown in southern Bahia. Cocoa beans from three cultivars were fermented and sun dried and monomeric phenolic compounds and methylxantines were determinated. The results showed that each cultivar have different amounts of phenolic compounds and the behaviour of them is different during fermentation. The amount of methylxantines varied but there was not a pattern for methylxantines behavior during process. In addition a huge reduction in phenolic compounds could be observed after drying. Differently of phenolic compounds, methylxantines did not have great modification after sun drying. So, the differences observed in this study between cultivars, take to the conclusion that the compounds studied in those cocoa cultivars have different behavior during fermentation and drying, which consequently, give to these cultivars differences in sensory characteristics.

Characterization of polyphenol oxidase in two cocoa (Theobroma cacao L.) cultivars produced in the south of Bahia, Brazil

Abstract The reactions leading to the formation of precursors of chocolate flavor are performed by endogenous enzymes present in the cocoa seed. Polyphenol oxidase (PPO) presence and activity during fermentation of cocoa beans is responsible for the development of flavor precursors and is also implicated in the reduction of bitterness and astringency. However, the reliability of cocoa enzyme activities is complicated due to variations in different genotypes, geographical origins and methods of fermentation. In addition, there is still a lack of systematic studies comparing different cocoa cultivars. So, the present study was designed to characterize the activity of PPO in the pulp and seeds of two cocoa cultivars, PH 16 and TSH 1188. The PPO activity was determined spectrophotometrically and characterized as the optimal substrate concentration, pH and temperature and the results were correlated with the conditions during the fermentation process. The results showed the specificity and differences between the two cocoa cultivars and between the pulp and seeds of each cultivar. It is suggested that specific criteria must be adopted for each cultivar, based on the optimal PPO parameters, to prolong the period of maximum PPO activity during fermentation, contributing to the improvement of the quality of cocoa beans.

Somatic embryogenesis and plant regeneration in elite clones of Theobroma cacao

The objective of this work was to evaluated a procedure for somatic embryogenesis and regeneration of cacao (Theobroma cacao L.) elite clones. Petal explants from cacao clones TSH 565 and TSH 1188 were cultured on PCG and SCG-2 media, for calli growth. Somatic embryos were formed on the surface of embryogenic calli after transfer to embryo development (ED) medium. Clone TSH 565 showed a higher embryogenic potential than TSH 1188. The best combination of carbon source for embryo induction in ED medium was genotype-specific. Embryogenic callus formations increased in micropore tape-sealed Petri dishes, irrespective of cacao genotype. Mature somatic embryos were successfully converted into plantlets.

Localization of the cotyledon reserves of Theobroma grandiflorum (Willd. ex Spreng.) K. Schum., T. subincanum Mart., T. bicolor Bonpl. and their analogies with T. cacao L.

Cotyledon mesophyll cell morphology and lipid and protein synthesis of T. grandiflorum, T. subincanum and T. bicolor were analyzed and compared with T. cacao. These species possess foliar cotyledons folded around the hypocotyl radicle axis, typical of Sterculiaceae. Fruit size, morphology and weight are very distinct amongst the four species and so are the respective seeds. The main axis of the T. grandiflorum and T. bicolor seeds measured about 30 mm, while T. subincanum and T. cacao seeds measured 17 mm and 26 mm respectively. The seed weights of T. grandiflorum, T. bicolor, T. subincanum and T. cacao were 11.6 g, 9.4 g, 2.1 g and 3.0 g, respectively. The cotyledon mesophylls of the four species contained mainly polysaccharides and lipid-protein reserve cells. Theobroma cacao, T. grandiflorum and T. subincanum were composed of greater than 50% lipids. For the four species, lipid globules gradually accumulated adjacent to the cell wall, and these globules measured from 1 to 3 µm. TEM showed low-density proteins inside the central vacuole of the young mesophyll cells of T. cacao. The protein reserves of the mature cells were densely scattered amongst the lipid bodies, and a few starch granules occurred together with the cotyledon mesophyll of the four species. Polyphenolic cells were found throughout the mesophyll cells or aligned with the respective vascular bundles. Immature cells demonstrated the capacity to synthesize all these reserves, but gradually the pre-determined cells produced mainly lipid-protein reserves. Besides the unique characteristics of the T. cacao products, the lipid-protein synthesis capacities of T. grandiflorum, T. subincanum and T. bicolor suggest various possibilities for new industrialized food, pharmaceutical and cosmetic products.

Efeitos de diferentes níveis de N, P e K sôbre cacaueiros jovens (Theorema cacao L.) em solução nutritiva

Cocoa seedlings were grown in nutrient solution in order to find out the effects of the levels of N, P, K in the substrate on growth, symptoms and chemical composition of the leaves. A 3x3x3 factorial design was used. The following conclusions can be drawn. 5.1. Among the three elements under study, only nitrogen had a positive, significant effect on growth as measured by height and dry weight. 5.2. The level of a given elements in the leaves did increase when its level in the nutrient solution was raised. On the other hand the N content was positively affected by the concentration of P in the substrate. 5.3. A decrease in the N/P and K/P rations in the leaves was observed when the nutrient solutions had, res-respectively, absence of K and N and high level both of K and N. The N/K ratio increased with the level of P in the substrate. 5.4 A highly significant positive correlation was found between N content in the leaves and dry weight. This was not the case however, insofar P and K are considered - no correlation whatsoever was observed.

Chemical, physico-chemical and sensory characterization of mixed açai (Euterpe oleracea) and cocoa´s honey (Theobroma cacao) jellies

Four formulations of mixed açaí (Euterpe oleracea) (A) and cocoa´s honey (Theobroma cacao) (CH) jellies were prepared according to the following proportions: T1 (40% A:60% CH), T2 (50% A:50% CH), T3 (60% A: 40% CH) and T4 (100% A - control). All formulations were prepared using a rate 60:40 (w/w) of sucrose and pulp, plus 0.5% pectin and the products reached to average of 65% soluble solids content. The jellies were analyzed by chemical and physicochemical (titratable acidity, pH, soluble solid content, dry matter, total protein, lipids, vitamin C and calories) and sensory characteristics; also were evaluated levels of P, K, Ca, Mg, Fe, Zn, Cu and Mn. It was used a hedonic scale of 7 points to evaluate the attributes: overall impression, spreadability, brightness, flavor, texture and color, and also was verified the purchase intention score. The titratable acidity and pH ranged from 0.46 to 0.64% and 3.35 to 3.64, respectively, that are within the range found at most fruit jellies. The soluble solids content ranged between 65.2 and 65.5 ºBrix. The sensory acceptance results showed that all treatments (T1, T2, T3 and T4) presented means of sensory attributes above 4, demonstrating good acceptance of the product, but the treatment T1 presented the higher scores for the evaluated attributes. Cocoa´s honey added a positive influence on the attributes of color, texture and spreadability.

GROWTH AND NUTRITION OF CACAO SEEDLINGS INFLUENCED BY ZINC APLICATION IN SOIL

ABSTRACT Levels of Zn in tropical soils profoundly influences growth and nutrition of tree crops. Research was undertaken to assess the effect of soil Zn on growth and nutrition of clonal cacao tree seedlings of PH 16. Three acidic Oxisol soils differing in texture were used with nine doses of Zn (0, 1, 2, 4, 8, 16, 32, 48, and 64 mg dm-3). Rooted clonal seedlings were grown in plastic pot with 11 dm-3 of the soils at varying Zn levels for 240 days. At harvest growth (dry matter mass of leaves, stems, shoots, roots, and total) and nutrient concentrations were determined. The clonal cacao seedlings showed differences for production of dry matter mass and foliar nutrient concentrations for P, K, Ca, Mg, Mn, Fe, Zn, and Cu. There was Zn toxicity in all soils.