A thermostable β-1,6-endoglucanase or pustulanase (E.C. 3.2.1.75) which catalyses the cleavage of β-1,6 bonds in pustulan and other beta-glucans containing 1,6 linked glucose units. The enzyme is now a component in a standard assay for beta-glucan from Baker’s yeast as documented in US Pharmacopeia Food and Chemicals Index 7th Edition, 3rd Supplement p. 1686-1689.

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Enzyme activity: Cel136 Beta-glucanase (glucan endo-1,6-β-glucosidase) hydrolyses Beta-glucan polysaccharides with β-(1,6) linkages – see further below for substrate specificity

Activity determination: The standard assay for activity was done by incubating the enzyme at 80°C for 10 min. with 1% (w/v) pustulan as substrate in 0,1 M sodium phosphate buffer at pH 5.5. 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, pustulanase, glucan endo-1,6-β-glucosidase, 6-β-D-glucan glucanohydrolase, endo-1,6-β-glucanase; β-1→6)-β-D-glucanase; β-1,6-glucanase-pustulanase; β-1,6-glucan hydrolase; β-1,6-glucan 6-glucanohydrolase; 1,6-β-D-glucan glucanohydrolase

Protein family: Glycosyl hydrolase family 30 ( GH30 ) – CAZy database family 30

Enzyme classification: EC 3.2.1.75

Source: Proprietary metagenome environmental DNA

Beta-Glucan Assay: Beta-glucanase (Pustulanase) Cel136 is recommended for the determination of yeast Beta-Glucan.

The United States Pharmacopeia (USP) and the USA-based company Biothera have developed the first official monograph for beta glucan from bakers yeast (Saccharomyces Cerevisiae), recognized for its immune health benefits and used as dietary supplement in food and beverages. The monograph was  issued in the Third Supplement of the Seventh Edition of the Food Chemicals Codex (compendium of standards for the purity and identity of food ingredients) and includes a protocol for beta-glucan assay using the Cel136 enzyme. See “Beta Glucan from Baker’s Yeast (Saccharomyces cerevisiae)”. Food Chemical Index, 7th Edition, Third supplement, p. 1686-1689. (2011)