Purification and determination of the action pattern of Haliotis tuberculata laminarinase

Abstract

The major laminarinase activity (EC 3.2.1.39) from the gastropodean marine mollusc Haliotis tuberculata was purified to homogeneity by cation exchange chromatography and its action pattern was investigated by HPAEC-PAD analysis of the degradation of various laminarin samples. It consists of a 60 kDa protein capable of depolymerizing the unbranched portions of the beta-(1-->3), beta-(1-->6)-glucan, down to laminaritriose. The enzyme operates via a molecular mechanism retaining the anomeric configuration. As the purified protein does not cleave the beta-(1-->6) linkages, it can be used for the structural analysis of laminarins.

Purification and characterisation of endo-β-1,4-glucanase and laminarinase enzymes from the gecarcinid land crab Gecarcoidea natalis and the aquatic crayfish Cherax destructor

“…This discrepancy suggests that laminarinase may exist as a dimer, joined by either a disulphide bond or van der Waals forces. Given that β-1,6-glycosidic bonds within laminarin cause the molecule to be branched (Lépagnol-Descamps et al, 1998), the laminarinase dimer may fit in between these branches. Simultaneous hydrolysis of the two branches may make the laminarinase more efficient.…”

Enzymatic properties and the primary structure of a β-1,3-glucanase from the digestive fluid of the Pacific abalone Haliotis discus hannai

“…Among these seaweeds' polysaccharides, laminarin seems to be the most potential glucose source for abalone since laminarin is a major storage polysaccharide consisting of -1,3-linked glucose main chains possessing occasional -1-6-linked glucose branches (Maeda and Nisizawa, 1967;Pang et al, 2005). Previously, a laminarin-degrading enzyme, i.e., endo--1,3-glucanase that hydrolyzes -1,3-glucoside linkages of laminarin, was isolated from hepatopancreas of the ormer Haliotis tuberculata (Lépagnol-Descamps et al, 1998). On the other hand, laminarin content in brown seaweeds has been shown to come up to approximately 20% (w/w, in dried materials).…”

“…-1,3-Glucanases from surf clam (Spisula sachalinensis), scallop (Chlamys albidus, Patinopecten yessoensis), and the ormer (H. tuberculata) have been classified to the first type enzyme (EC 3.2.1.6). These marine molluscan enzymes are also called laminarinase since they can efficiently hydrolyze the -1,3-linkage of laminarin from brown seaweeds (Sova et al, 1970b;Privalova and Elyakova, 1978;Lépagnol-Descamps et al, 1998;Kumagai et al, 2008). Primary structures of molluscan -1,3-glucanases have been reported in the surf clam and scallop enzymes (Kozhemyako et al, 2004;Kovalchuk et al, 2006;Kovalchuk et al, 2009).…”

“…Previously, Lépagnol-Descamps et al (1998) isolated a -1,3-glucanase from hepatopancreas of the ormer H. tuberculata (a gastropod), and investigated its basic properties.…”

Isolation and characterization of two types of β-1,3-glucanases from the common sea hare Aplysia kurodai

“…AkLam33 appeared to be the first exolytic β-1,3-glucanase from marine invertebrates since all β-1,3-glucanases so far purified from marine invertebrates have been characterized as endolytic enzymes (Sova et al, 1970;Lépagnol-Descamps et al, 1998;Kovalchuk et al, 2006;Kumagai et al, 2008;Pesentseva et al, 2008;Zhu et al, 2008;Kumagai and Ojima, 2009;Kovalchuk et al, 2009). Besides AkLam33, exolytic β-1,3-glucanases have been reported on bacteria, yeast, insect, and a terrestrial snail (Marshall and Grand, 1975;Sánchez et al, 1982;Kulminskaya et al, 2001;Hrmova and Fincher, 1998;Igarashi et al, 2003;Genta et al, 2007); however, these enzymes were classified under GHF 3, 5, 17, and 55 (Klebl and Tanner, 1989;Chambers et al, 1993;Hrmova et al, 1996;Sakamoto et al, 2005;Ishida et al, 2009;Ooi et al, 2009).…”