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Nucleic Acid Purification

GE013.0001 - Zymolyase® 20T, 1g

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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.
 

Features

  • Form: Lyophilized powder 
  • Purification: Ammonium Sulfate Precipitation
  • Activity: ß-1.3-glucan laminaripentaohydrolase (20 U/g)
  • Other activities: ß-glucanase (~1.5x106 U/g), protease (~1.0x104 U/g), mannanase (1.0x106 U/g)
  • Contaminants: trace amounts of amylase and xylanase
  • Optimum conditions for lysis of viable yeast cells: pH 7.5, 35?C
  • Optimum conditions for hydrolysis of yeast glucan: pH 6.5, 45?C
 

Applications

  • Preparation of spheroplasts or protoplasts from a variety of yeast strains
 

Contents

  • Zymolyase® 20T, lyophilized powder, 1g