About Zymolyase - Yeast Lytic Enzyme
Digestion of yeast and fungal cell walls is necessary for many experimental procedures including spheroplasting, immunofluorescence, transformation, protein purification, and others. The use of lytic enzymes like Zymolyase is routinely used for digestion. The Zymolyase from Zymo Research is prepared from Arthrobacter luteus, lyophilized, and packaged with a resuspension buffer. The buffer has been optimized to confer maximal levels of enzymatic activity. The main activities of the enzyme are β-1,3 glucanase and β-1,3-glucan laminaripentao-hydrolase, which hydrolyze glucose polymers at the β-1,3-glucan linkages releasing laminaripentaose as the principal product. Optimal Zymolyase activity is at 30° - 37°C; lytic activity ceases at higher temperatures. R-Zymolyase includes 0.5 U/µl RNase A when reconstituted. Susceptible fungal genera: Asbya, Candida, Debaryomyces, Eremothecium, Endomyces, Hansenula, Hanseniaspora, Kloekera, Kluyveromyces, Lipomyces, Metschikowia, Pichia, Pullularia, Saccharomyces, Saccharomycodes, Saccharomycopsis, Schizosaccahromyces, Torulopsis.
| Format | Lyophilized enzyme provided w/ storage buffer |
|---|---|
| Concentration | 5 U/µl |
| Source | Arthrobactor luteus |
| Storage | Zymolyase is stable for over 1 year at -20°C or many years below -70°C. |
| Unit Definition | One lytic unit is defined as catalyzing a 10% decrease in optical density at 800 nm (OD800) at 30°C in 30 minutes. |
| Optimum pH and Temperature | pH 7.5, 35°C (lysis of viable yeast), pH 6.5, 45°C (hydrolysis of yeast glucan). |
| Activity | ß-1,3-glucan laminaripentaohydrolase and ß-1,3-glucanase (trace amount of protease, ca. 1.5 units per 10 µl). DNase and RNase: none detectable. |
| Protein Content | ~10-15 mg/ml |
| Deactivation | Lytic activity is lost in 5 minutes at 60°C. |
| Assay Condition | 50 mM potassium phosphate, pH 7.5, 10 mM 2-mercaptoethanol in 1 ml yeast cell suspension of 0.8 - 1.0 OD800. |
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Zymolyase can be used for enzymatic digestion of yeast glycan coats and for spheroplast formation. The arrow indicates the nucleus and intracellular components of a spheroplast through a partially digested plasma membrane.*
*Source: A protocol for isolation and visualization of yeast nuclei by scanning electron microscopy (SEM). Elena Kiseleva, Terry D Allen, Sandra A Rutherford, Steve Murray, Ksenia Morozova, Fiona Gardiner, Martin W Goldberg & Sheona P Drummond. Nature Protocols 2, 1943 - 1953 (2007) Published online: 9 August 2007 doi:10.1038/nprot.2007.251 |
