Zymolyase® 20T is prepared from Arthrobacter luteus and is supplied as an ammonium sulfate precipitate of a complex of enzymes in the form of a lyophilized powder. The strong lytic activity against living yeast cell walls is mainly due to the activity of ß-1,3-glucan laminaripentaohydrolase. This enzyme hydrolyzes linear glucose polymers with β-1,3-linkages and releases laminaripentaose as the main and minimum product unit. This allows the production of protoplasts or spheroplasts of various yeast strains.
The lytic activity of Zymolyase® 20T varies depending on the yeast strain, growth stage of the yeast, or cultural conditions. Under the conditions described below, the lytic activity of Zymolyase® 20T is 20 U/g.
One unit (U) of lytic activity is defined as the amount that results in a 30% decrease of the absorbance at 800nm of a reaction mixture* after incubation with gentle shaking at 25ºC for 2 hours. A decrease of A800 with 60%, equivalent to 2U, corresponds to complete lysis. In other words, 1U of Zymolyase® lyses 3mg of dry weight Saccharomyces cerevisiae in 2 hours under ideal conditions.
*10ml reaction mixture is composed of 1 ml of distilled water, 3 ml of S. cerevisiae (2mg/ml), 5 ml 1/15 M phosphate buffer (pH 7.5), and 1 ml of enzyme solution.
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