Description
Zymolyase® 20T from GRiSP is prepared from Arthrobacter luteus, and is supplied as an ammonium sulfate precipitate of a complex of enzymes. 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 specifically laminaripentaose as the main and minimum product unit, allowing for the production of protoplasts or spheroplasts of various yeast strains.
Lytic activity varies depending on yeast strain, growth stage of yeast, or cultural conditions. Under the conditions mentioned below, the lytic activity of Zymolyase® 20T is 20.000 U/g,
Applications
Preparation of spheroplasts or protoplasts from a variety of yeast strains.
Features
Unit definition *
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 S. cerevisiae in 2 hours under ideal conditions.
*) 10ml reaction mixture is: 1 ml of distilled water, 3 ml of Saccharomyces cerevisiae (2mg/ml), 5 ml 1/15 M phosphate buffer (pH 7,5), and 1 ml of enzyme solution.
Specifications
Form: Lyophilized powder
Purification: Ammonium Sulfate Precipitation
Activity:
ß-1,3-glucan laminaripentaohydrolase: 20.000 U/g
Other Activities
ß-glucanase: ~1.5×106 U/g
protease: ~1.0×104 U/g
mannanase: ~1.0×106 U/g
Contaminants
Amylase: trace amounts
Xylanase: trace amounts
Optimum Conditions (pH, temp)
For lysis of viable yeast cells: pH 7.5 – 35ºC
For hydrolysis of yeast glucan: pH 6.5 – 45ºC
Storage/Stability
+2ºC to +8ºC: No loss of activity after storage for 1 year.
+30ºC: 70% of lytic activity is lost after 3 months.
+60ºC: All lytic activity is lost after 5 minutes.
Specificity
Lytic Spectrum: Ashbya, Candida, Debaryomyces, Eremothecium, Endomyces, Hansenula, Hanseniaspora, Kloeckera, kluyveromyces, Lipomyces, Metschnikowia, Pichia, Pullularia, Saccharomyces, Saccharomycopsis, Schwanniomyces, Torulopsis, Saccharomycodes, etc.
