A highly thermostable Laminarinase (Beta-glucanase). The enzyme is active on β-1-3 linkages in polysaccharides of D-glucose units with β-1-3 linkages including β-glucans with mixed linkages such as β-1-3 and β-1-4 or β-1-3 and β-1-6 linkages includinglaminarin, lichenan, barley beta-glucan and scleroglucan. Uniquely active on recalcitrant beta-glucans such as scleroglucanand curdlan.
The enzyme is available for purchase as 200 units size (see unit definition below).
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Activity determination: The standard assay for activity was done by incubating the enzyme at 75°C for 15 min. with 1% (w/v) lichenan as substrate in 0,1 M sodium phosphate buffer at pH 7.0. The reducing sugars released were detected by the dinitrosalisylic acid method using glucose as standard.
Unit definition: One unit (U) of enzyme activity is the amount that leads to the release of 1 μmol reducing sugars per minute.
Synonyms: glycoside hydrolase, 1,3-[1,3;1,4]-β-D-Glucan-3(4)-glucanohydrolase, Endo-1,3(4)-β-glucanase,laminaranase, lichenase, licheninase.
Protein family: Glycosyl hydrolase family 16 ( GH16 ) – CAZy database
Enzyme classification: EC 22.214.171.124
Source: The thermophilic bacteria Rhodothermus marinus
Protein sequence: NBCI protein entry
Structural information: The crystal structure of Laminarinase from Rhodothermus marinus (96% sequence identity with Bglu110 Laminarinase/Lichenase) has been determined to 1.9 Å resolution. – PDB entry 3ILN
Crystal structure of Laminarinase from Rhodothermus marinus
The molecule laminarin (also known as laminaran) is a storage glucan produced in brown algae through photosynthesis. The polysaccharide is made up of glucose residues withβ-1,3-linkages and β-1,6-linkages. It is a linear polysaccharide, with a β(1→3):β(1→6) ratio typically 3:1 but the ratio may vary with the source of the polysaccharide.
Scleroglucan, is formed by the fungus Sclerotium rolfsii. A chemically analogous polysaccharide, Schizophyllan (Sizofiran, Sonifilan, SPG) is a neutral extracellular polysaccharide produced by the fungus Schizophyllum commune. Schizophyllan is a β-1,3 beta-glucan with β-1,6 branching. Both polysaccharides share the chemical structure of the backbone with curdlan.
Properties of Bglu110 Laminarinase/Lichenase (β-glucanase):
pH optimum: the enzyme has optimum activity around pH 7
Thermostability: Activity of the enzyme as a function of time at different temperature