Purificação e caracterização de bromelina a partir do extrato bruto de Ananas comosus por adsorção em leito expandido / Purification and characterization of bromelain, from crude extract of Ananas comosus, using expanded bed adsorption

AUTOR(ES)
DATA DE PUBLICAÇÃO

2007

RESUMO

Bromelain are extracted from Ananas comosus infrutescence, and are used in food industry, as a meat softener, and pharmaceutical industry, as well. Bromelain are presented in the early stages of fruit development, however, their levels increase quickly, remaining high until the maturation. Expanded Bed Adsorption (EBA) is a chromatography technique for separation and purification of biological products directly from crude feedstock, without using centrifugation, microfiltration and other clarification prior steps. This technique permits crude feeding into chromatographic column without initial treatment, and as the bed expands, it increases adsorbent surface contact, making interaction with the target molecule more effective. This work reports the adsorption of Bromelain in expanded bed conditions, such as the adsorption kinetics parameters, the bed height, and the flow velocity. This work also focus the partial characterization of bromelain, describing aspects such temperature effect, pH effect, and its influences within time. It was observed a growing enzymatic activity by increasing the temperature level, and with the proximity to neutral pH. It was also observed that increments in temperature over 65 ºC cause a little decrease of enzymatic levels. After 150 minutes, brmoelain retained 50 % of its initial activity. The maximum adsorption capacity was determined by the adsorption isotherm, where the Qm and kd values could be estimated, 9.18 U/mL and 0.591 U/mL, respectively. The resident time was reduced 10 fold by increasing the expansion degree 2.5 times; nonetheless the plate value (N) was reduced only 2 fold. The purification factor was about 13 fold. EBA showed to be feasible for purification of bromelain, and the eluted enzyme as SDS-PAGE

ASSUNTO(S)

enzimas - purificação adsorption adsorção mo leito fluidizado p bromelains ensymes purification bromelina

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