Efficient degradation of yeast cell walls
The digestion of fungal and yeast cell walls is a necessary part of the protocol for many experimental procedures including immunofluorescence, transformation, protein purification and spheroplasting. To achieve this digestion, lytic enzymes such as Zymolyase are routinely used.
AMSBIO offers Zymolyase, produced by a submerged culture of Arthrobacter luteus1, which has an effective lytic activity against cell walls of viable yeast cells2,3 to produce protoplast or spheroplast of various strains of yeast cells. An essential enzyme for the lytic activity of Zymolyase is β-1,3-glucan laminaripentaohydrolase. It hydrolyzes linear glucose polymers at β -1,3-linkages and releases specifically laminaripentaose as the main and minimum product unit4,5,10,11.
- Convenience: Provided as a lyophilized powder and reconstituted with a buffer solution
- Very efficient cell wall digestion
- Prepared from Arthrobacter luteus, their essential enzyme activity is β-1,3-glucan laminaripentao-hydrolase
- Protoplast/spheroplast preparation
- Yeast cell fusion
- Transformation of yeast cells
- Yeast genetics
There are two preparations of Zymolyase: Zymolyase-20T and Zymolyase-100T, having lytic activity of 20,000 units/g and 100,000 units/g respectively. Zymolyase-20T is ammonium sulfate precipitate while Zymolyase-100T is a further purified preparation by affinity chromatography9. Lytic activity varies depending on yeast strain, growth stage of yeast, or cultural conditions. Further information related to Zymolyase® can be obtained in the reference section below12-16.
|Purification||Zymolyase® -20T:(NH4)2SO4 precipitation Zymolyase® -100T:Affinity Chromatography|
|Activity||Zymolyase® -20T:20,000 units/gram Zymolyase® -100T:100,000 units/gram|
|Essential enzyme||β.-1, 3-glucan laminaripentaohydrolase|
|Other activities contained||Zymolyase® - 20T||Zymolyase® - 100T|
|B-1, 3-glucanase:||ca. 1.5 x 106 units/g||ca. 1.0 x 107 units/g|
|protease:||ca. 1.0 x 104 units/g||ca. 1.7 x 104 units/g|
|mannanase:||ca. 1.0 x 106 units/g||ca. 6.0 x 104 units/g|
|Contaminants:||Trace amounts of amylase, xylanase, phosphatase No DNase, RNase detected|
|Heat stability||The lytic activity is lost on incubation at 60°C for 5 minutes|
|Specificity (lytic spectrum):||Ashbya, Candida, Debaryomyces, Eremothecium, Endomyces, Hansenula, Hanseniaspora, Kloeckera, Kluyveromyces, Lipomyces, Metschikowia, Pichia, Pullularia, Torulopsis, Saccharomyces, Saccharomycopsis, Saccharomycodes, Schwanniomyces, etc.|
|Activators:||SH compound such as cystein, 2-mercaptoethanol of dithiothreitol|
|Stability:||No loss of activity was found after storage for 1 year at 4°C|
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