| Zymolyase, produced by a submerged culture of Arthrobacter luteus (1), is an enzyme preparation which effectively lyses cell walls of viable yeast cells. An essential enzyme responsible for lysis of viable yeast cells in this preparation is b-1, 3-glucan laminaripentaohydrolase. It hydrolyzes linear glucose polymers with b-1,3-linkages and releases laminaripentaose specifically as the main and minimum product unit (4,5,10,11). This lytic activity releases spheroplasts and protoplasts in the preparation of yeast DNA prior to restriction enzyme digestion and Southern Blot analysis. The extent of lysis of yeast cells by Zymolyase varies with yeast strain, growth stage of yeast and cultural condition (6-8). Further information related to Zymolyase is obtained in the references below. Zymolyase 100T Stock Concentrate 10mg/ml is a convenient preparation for use in yeast spheroplast/protoplast fusion and transformation protocols. Also useful for yeast tetrad dissection solutions. Applications: Protoplast/Spheroplast Preparation Yeast Cell Fusion Yeast Cell Transformation Appearance: Clear to pale yellow solution/suspension Activity: 100u/mg Purity: Purified by affinity chromotography. Unit Definition: One unit of lytic activity is defined as that amount which indicates 30% of decrease in absorbance at 800nm (A800). Refer to Assay for Enzymatic Activity. Water: Molecular Biology Grade, RNase, DNase-Free Filter: 0.2um validated Sorbitol: 98% Synonyms: Lyticase*, Yeast Lytic Enzyme Source: Arthrobacter luteus Essential Enzyme: b-1,3-glucan laminaripentaohydrolase Contaminants: Certified to be free of contaminating ribonuclease and deoxyribonuclease. Other activities contained: b-1,3-glucanase: ~1.5x10e3u/mg Protease: ~15u/mg Mannanase: ~1.5x10e3u/mg Amylase, xylanase, phosphatase: Minute amounts DNase, Rnase: Trace levels reported (See reference No. 3 as to the definition of each enzyme unit. Each activity varies more of less among lots.) Optimum pH: For cell wall lysis: 7.5, 35C For yeast glucan hydrolysis: 6.5, 45C Stable pH: 5-10 Heat Stability: The lytic activity is lost on incubation at 60C for 5 minutes. Properties of Zymolyase: Specificity (lytic spectrum)(5): Ashbya, Candida, Debaryomyces, Eremothecium, Endomyces, Hansenula, Hanseniaspora, Kloekera, Kluyveromyces, Lipomyces, Metschikowia, Pichia, Pullularia, Torulopsis, Saccharomyces, Saccharomcopsis, Saccharomycodes, Schwanniomyces, etc Susceptible strains in low concentration (0.2 units/ml): Ashbya, Endomyces, Kloekera, Kluyveromyces, Pullularia, Saccharomyces Susceptible strains in high concentration (2.0 units/ml): Candida, Debaryomyces, Eremothecium, Hansenula, Hanseniaspora, Lipomyces, Metschnikowia, Saccharomycopsis, Saccharomycodes, Schizosaccharomyces, Selenozyma, Trigonopsis, Wickerhamia Susceptibility depending upon strains: Bretanomyces, Cryptococcus, Nadsonia, Picia, Rodosporidium, Schwanniomyces, Stephnoascus, Torulopsis No susceptible strains: Bullera, Pityosporum, Rhosotorula, Sporidiobolus, Sporobolomyces, Stetigmatomyces, Trichosporon Activators: SH compound such as cystein, 2-mercaptoethanol of dithiothreitol Storage and Stability: Stable for 12 months after receipt at -20C. Companion Products: Z1000: Zymolyase 20T Z1001: Zymolyase 20T Concentrate 50mg/ml, 0.1M Sorbitol Zymolyase 20T shows 20u/mg of the lytic activity, defined after, toward brewers yeast cells (Saccharomyces cerevisiae, resting stage) or toward yeast cells of Saccharomyces cerevisiae IFO 0565 cultured statically in malt extract medium (Malt extract 2g, peptone 0.5g, water 100ml) at 20C for 34 hours. Z1004: Zymolyase 100T Z1005: Zymolyase 100T Concentrate 10mg/ml, 0.1M Sorbitol Zymolyase 100T, further purified, whose specific activity is 100u/mg. Note: Zymolyase100T is less soluble than 20T and may not be completely dissolved in buffers. If this is the case use as a suspension. |
