Please cite this article in press as: Perrella, N.N., et al., Characterization of -L-fucosidase and other digestive hydrolases from Biom- phalaria glabrata. Acta Trop. (2014), http://dx.doi.org/10.1016/j.actatropica.2014.08.022 ARTICLE IN PRESS G Model ACTROP-3451; No. of Pages 10 Acta Tropica xxx (2014) xxx–xxx Contents lists available at ScienceDirect Acta Tropica jo ur nal home p age: www.elsevier.com/locate/actatropica Characterization of -L-fucosidase and other digestive hydrolases from Biomphalaria glabrata Natalia N. Perrella a,b , Rebeca S. Cantinha c,d , Eliana Nakano c , Adriana R. Lopes a,∗ a Laboratory of Biochemistry and Biophysics—Instituto Butantan, São Paulo, Brazil b Programa de Pós Graduac ¸ ão Interunidades em Biotecnologia PPIB, Universidade de São Paulo, São Paulo, SP, Brazil c Laboratory of Parasitology—Instituto Butantan, São Paulo, Brazil d Instituto de Pesquisas Energéticas e Nucleares, Universidade de São Paulo, São Paulo, SP, Brazil a r t i c l e i n f o Article history: Received 10 February 2014 Received in revised form 3 July 2014 Accepted 12 August 2014 Available online xxx Keywords: Hepatopancreas Enzymes -L-Fucosidase Fucoidan Biomphalaria glabrata Schistosoma mansoni a b s t r a c t Schistosoma mansoni is one of the major agents of the disease Schistosomiasis, which is one of the major global public health concerns. Biomphalaria glabrata is an obligate intermediate mollusc host of S. mansoni. Although the development of S. mansoni occurs in the snail hepatopancreas, studies that focus on this organ remain limited. In this study, we biochemically identified five distinct carbohy- drases (amylase, maltase, -glucosidase, trehalase, and -L-fucosidase), lipases, and peptidases in the B. glabrata hepatopancreas and focused on the isolation and characterization of the activity of -L- fucosidase. The isolated -L-fucosidase has a molecular mass of 141 kDa, an optimum pH of 5.8, and is inhibited by Tris, fucose, and 1-deoxyfuconojirimycin. B. glabrata -L-fucosidase is an exoglycosidase that can hydrolyze the natural substrate fucoidan to fucose residues. It presented K m values of 48.4 M to 4-Methylumbelliferyl -L-fucopyranoside and 0.55 mM to p-nitrophenyl--L-fucopyranoside. Thus, - L-fucosidase has a high activity in the hepatopancreas of B. glabrata, and the differential expression of this enzyme between susceptible and resistant strains indicates that besides its digestive role, -L-fucosidase may also be important in host/parasite interactions. © 2014 Elsevier B.V. All rights reserved. 1. Introduction Schistosomiasis, a serious public health problem, is second only to malaria (WHO, 2013; http://www.who.int). Globally, 200 million people are infected with the snail-transmitted, water-borne par- asitic helminth Schistosoma mansoni, and severe infections claim 20,000 lives annually. Multiple studies show that during infection in mammalian or mollusc hosts, the different life-cycle stages of S. mansoni present a series of glycoconjugates that play impor- tant roles in the host-parasite interplay (Van Die et al., 2010). For example, glycosphingolipids extracted from cercariae and eggs are known to contain a series of fucose residues (Weiss et al., 1986; Wuhrer et al., 2002). Abbreviations: MUB, 4-methylumbelliferyl butyrate; DMPTB, dimercapto- 1-propanol tributyrate; MUFUC, 4-methylumbelliferyl -L-fucopyranoside; MUGLU, 4-methylumbelliferyl -D-glucopyranoside; NPFUC, p-nitrophenyl -L-fucopyranoside; BgFUC, -L-fucosidase from Biomphalaria glabrata. ∗ Corresponding author. Tel.: +55 11 2627 9746. E-mail addresses: adriana.lopes@butantan.gov.br, dririoslopes@gmail.com (A.R. Lopes). Studies on S. mansoni and the snail Biomphalaria glabrata have focused mainly in Biomphalaria’s hemocytes and defense mecha- nisms. In snails, hepatopancreas is an organ that provides the most favorable conditions for the development and multiplication of parasites by providing abundant food supply to sporocysts (Becker, 1980). Studies that focus on the hepatopancreatic environment as an interface for parasite-host interactions remain limited. Recently, Myers et al. (2008) investigated the role of proteolytic enzymes from B. glabrata in the development of the S. mansoni and its dis- tribution in the hepatopancreas, ovotestis, albumen gland, and cell-free plasma (hemolymph). The authors also identified clones corresponding to B. glabrata cellulase, elastase, disintegrin and met- allopepetidase, lysozyme, -L-fucosidase, and serine peptidase in the EST data. The dependence of the fucose residues present on glycoconjugates on host-parasite interactions and the evidence of a possible hepatopancreatic -L-fucosidase that can catalyze the removal of these fucose residues, led us to hypothesize that this - L-fucosidase may facilitate the removal of the fucose residues from the glycosphingolipids present in S. mansoni in order to diminish host/parasite interaction. -L-fucosidase (EC 3.2.1.51) is a ubiquitous lysosomal glycoside hydrolase (GH) from family 29, (Shaikh et al., 2013) and catalyzes http://dx.doi.org/10.1016/j.actatropica.2014.08.022 0001-706X/© 2014 Elsevier B.V. All rights reserved.