Encapsulated Microparticles of (1→6)-ß-d-Glucan Containing Extract of Baccharis dracunculifolia: Production and Characterization.

ß-Glucans are biomacromolecules well known, among other biological activities, for their immunomodulatory potential. Similarly, extracts of Baccharis dracunculifolia also possess biological properties and are used in folk medicine for the treatment of inflammation, ulcers, and hepatic diseases. Microparticles containing (1→6)-ß-d-glucan (lasiodiplodan) and B. dracunculifolia extract were produced and characterized. A 23 factorial design was employed to define the conditions of production of microparticles by atomization. Lasiodiplodan associated with maltodextrin and gum arabic was studied as a matrix material. Microparticles of 0.4 µm mean size and high phenolics content (3157.9 µg GAE/g) were obtained under the optimized conditions. The microparticle size ranged from 0.23 to 1.21 µm, and the mathematical model that best represented the release kinetics of the extract was the Korsmeyer-Peppas model. Diffusional exponent (n) values of 0.64 at pH 7.7 and 1.15 at pH 2.61 were found, indicating particles with a non-Fickian or anomalous transport system, and Super Case II transport, respectively. Thermal analysis indicated that the microparticles demonstrated high thermal stability. The X-ray diffraction analyses revealed an amorphous structure, and HPLC-DAD analysis showed microparticles rich in phenolic compounds: caffeic acid, p-coumaric acid, and catechin. The microparticles obtained comprise a new biomaterial with biological potential for applications in different fields.

Fungal Exocellular (1-6)-ß-d-glucan: Carboxymethylation, Characterization, and Antioxidant Activity.

Exocellular (1→6)-ß-d-glucan (lasiodiplodan) produced by the fungus Lasiodiplodia theobromae MMPI was derivatized by carboxymethylation using different concentrations of a derivatizing agent. Lasiodiplodan was derivatized by carboxymethylation in an attempt to increase its solubility and enhance its biological activities. Carboxymethylglucans with degrees of substitution (DS) of 0.32, 0.47, 0.51, 0.58, and 0.68 were produced and characterized. FTIR analysis showed a band of strong intensity at 1600 cm-1 and an absorption band at 1421 cm-1, resulting from asymmetric and symmetrical stretching vibrations, respectively, of the carboxymethyl group COO- in the carboxymethylated samples. Thermal analysis showed that native lasiodiplodan (LN) and carboxymethylated derivatives (LC) exhibited thermal stability up to 200-210 °C. X-ray diffractometry demonstrated that both native and carboxymethylated lasiodiplodan presented predominantly an amorphous nature. Scanning electron microscopy revealed that carboxymethylation promoted morphological changes in the biopolymer and increased porosity, and alveolar structures were observed along the surface. The introduction of carboxymethyl groups in the macromolecule promoted increased solubility and potentiated the hydroxyl radical-scavenging activity, suggesting a correlation between degree of substitution and antioxidant activity.

Screening of Colletotrichum isolates from the Amazon biodiversity as producers of laccases using a simple methodology / Seleção de isolados de Colletotrichum da biodiversidade da Amazônia como produtores de lacases utilizando uma metodologia simplificada

This work presents a simple and rapid technique to select fungal strains with the ability to produce laccases. Fifteen strains of Colletotrichum spp. from the Amazon region (isolated from cumarú, puerária, cupuaçu and weed plants), as well as a strain isolated from espinheira santa in the south of Brazil, were found to produce laccases. Laccase activity was assayed using ABTS and DMP as putative substrates. Only Colletotrichum isolates of puero (P2 and P3) did not present activity towards DMP. Isolates D3 (weed plants), M3 (espinheira santa), C2 and C3 (cumarú) were selected, and were compared with the basidiomycete Pleurotus ostreatus used as a reference laccase producer under stationary and submerged culture conditions. Highest laccase activity was detected only in the stationary cultures of Colletotrichum isolate D3 assayed against ABTS (35 U/l) and DMP (8 U/l).

O presente trabalho apresenta uma metodologia simples e rápida para selecionar isolados fúngicos produtores de lacases. Foram avaliados quinze isolados de Colletotrichum da biodiversidade da Amazônia (obtidos de cumarú, puerária, cupuaçu e plantas daninhas), além de um isolado de espinheira santa do sul do país, quanto à produção de lacases. A atividade da enzima foi determinada utilizandose os substratos ABTS e o DMP. Somente os isolados de puerária P2 e P3 não apresentaram atividade sobre o substrato DMP. Os isolados D3 (planta daninha), M3 (espinheira santa), C2 e C3 (cumarú) foram selecionados e comparados quanto à produção de lacases em cultivos líquidos estáticos e agitados, utilizando-se o Pleurotus ostreatus, um basidiomiceto produtor de lacase, como referência. A maior atividade de lacase foi detectada apenas nos cultivos estáticos do isolado D3 (35U/l em ABTS; 8U/l em e DMP).

Screening of Colletotrichum isolates from the Amazon biodiversity as producers of laccases using a simple methodology / Seleção de isolados de Colletotrichum da biodiversidade da Amazônia como produtores de lacases utilizando uma metodologia simplificada

This work presents a simple and rapid technique to select fungal strains with the ability to produce laccases. Fifteen strains of Colletotrichum spp. from the Amazon region (isolated from cumarú, puerária, cupuaçu and weed plants), as well as a strain isolated from espinheira santa in the south of Brazil, were found to produce laccases. Laccase activity was assayed using ABTS and DMP as putative substrates. Only Colletotrichum isolates of puero (P2 and P3) did not present activity towards DMP. Isolates D3 (weed plants), M3 (espinheira santa), C2 and C3 (cumarú) were selected, and were compared with the basidiomycete Pleurotus ostreatus used as a reference laccase producer under stationary and submerged culture conditions. Highest laccase activity was detected only in the stationary cultures of Colletotrichum isolate D3 assayed against ABTS (35 U/l) and DMP (8 U/l).

O presente trabalho apresenta uma metodologia simples e rápida para selecionar isolados fúngicos produtores de lacases. Foram avaliados quinze isolados de Colletotrichum da biodiversidade da Amazônia (obtidos de cumarú, puerária, cupuaçu e plantas daninhas), além de um isolado de espinheira santa do sul do país, quanto à produção de lacases. A atividade da enzima foi determinada utilizandose os substratos ABTS e o DMP. Somente os isolados de puerária P2 e P3 não apresentaram atividade sobre o substrato DMP. Os isolados D3 (planta daninha), M3 (espinheira santa), C2 e C3 (cumarú) foram selecionados e comparados quanto à produção de lacases em cultivos líquidos estáticos e agitados, utilizando-se o Pleurotus ostreatus, um basidiomiceto produtor de lacase, como referência. A maior atividade de lacase foi detectada apenas nos cultivos estáticos do isolado D3 (35U/l em ABTS; 8U/l em e DMP).

Genetic diversity among Botryosphaeria isolates and their correlation with cell wall-lytic enzyme production

Nine isolates of Botryosphaeria spp. were evaluated for their growth and the production of cell wall-lytic enzymes (laccase, pectinase and beta-1,3-glucanase) when grown on basal medium in the absence and presence of the laccase inducer, veratryl alcohol (VA). The genetic relationship among the nine isolates collected from different host plants was determined by RAPD analyses. ITS sequence analysis showed eight closely related isolates classified as Botryosphaeria rhodina, and one isolate classified as Botryosphaeria ribis. RAPD analysis resolved the isolates into three main clusters based upon levels of laccase and beta-1,3-glucanase activity. There appears to be no correlation between pectinase production and genetic diversity among the nine isolates. However, the strain characterized as B. ribis, positioned out of the main cluster, was found to be the highest producer of pectinases in the presence of VA.

Nove isolados de Botryosphaeria spp foram avaliados quanto ao crescimento e produção de enzimas líticas da parede celular (lacase, pectinase e beta-1,3-glucanase) quando cultivados em meio basal na ausência e presença do indutor de lacase álcool veratrílico (VA). As relações genéticas entre os nove isolados coletados de diferentes plantas hospedeiras foram determinadas por RAPD. A análise das seqüências de nucleotídeos da região ITS mostrou oito isolados estreitamente relacionados, os quais foram classificados como Botryosphaeria rhodina e um isolado como Botryosphaeria ribis. A análise por RAPD agrupou os isolados em três grupos principais condizentes com os níveis de atividades de lacase e beta-1,3-glucanase. Nenhuma correlação foi detectada entre a produção de pectinase e a diversidade genética nos nove isolados. Entretanto, a linhagem caracterizada como B. ribis, posicionada fora dos grupos principais, se mostrou maior produtora de pectinase na presença de álcool veratrílico.