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.
![reaction of Zymolyase](http://www.nacalai.co.jp/global/reagent/images/REACTION.gif)
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.