Hidrolases do suco digestivo do molusco-praga Achatina fulica

Abstract

Achatina fulica, popularly known as the African snail, is a pulmonary terrestrial mollusc that is currently on the list of the 100 worst invasive species in the world, being found in almost all continents, causing environmental imbalance. The digestive juice of Achatina fulica is a very attractive source of enzymes from a biotechnological point of view. This work aimed to characterize the digestive juice (SDAf) as to its enzymatic diversity and to induce through specific feeding, the production of enzymes of interest, for example alkaline phosphatase, to later carry out the purification of this enzyme by the foaming methodology , or foaming adsorption, compared to other known techniques. Total protein amounts of SDAf were determined by four distinct methods Bradford, BCA, Biureto and Lowry, the latter being determined as standard. The following activities were found in relation to the polysaccharide substrates: rice amylopectin, apple arabana, cherry coffee arabinogalactan, bacterial cellulose, P. taeda cellulose-Kfaft, guar gum, corn phytoglycogen, bagasse hemicellulose, inulin, Neurospora cell walls citrus pectin, pullulan, tamarind xyloglucan. However, the dietary induction of the subjects did not provide significant differences in the enzymatic levels when the substrates were inulin or pectin. High enzymatic activities correlated with glycosidases of the disaccharidases type were found against the pNF-glucoside substrates. The highest levels of specific activities were for β - Glucosidase, β - Glucuronidase and β - Galactosidase. The same was observed for a commercially available enzyme, alkaline phosphatase (ALP), to hydrolyze with the substrate pNFF. The presence of ALP in the SDAf was confirmed by a zymogram. Purification for ALP with ion exchange chromatography columns and fractionation by ammonium sulfate precipitation did not provide good results, which was achieved through a foaming column. In comparison with the native enzyme, the inclusion of saponin at 0.25 mg / mL allowed for a 19% enrichment.