Zymolyase 100T (Lyticase, Yeast Lytic Enzyme)

Blue Ice

Molecular Biology Grade

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.

Applications:
Protoplast/Spheroplast Preparation
Yeast Cell Fusion
Yeast Cell Transformation

Appearance:
Lyophilized powder

Activity:
≥100U/mg

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.

Synonyms:
Lyticase*, Yeast Lytic Enzyme

Source:
Arthrobacter luteus

Essential Enzyme:
b-1, 3-glucan laminaripentaohydrolase

Contaminants:
DNase, Rnase: Trace levels detected.

Other Activities Contained:
b-1, 3-glucanase: ~1.5x10e3u/mg
Protease: ~15u/mg
Mannanase: ~1.5x10e3u/mg
Amylase, xylanase, phosphatase: Minute amounts
(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, 35°C
For yeast glucan hydrolysis: 6.5, 45°C

Stable pH:
5-10

Heat Stability:
The lytic activity is lost on incubation at 60°C 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
Non-susceptible strains:
Bullera, Pityosporum, Rhosotorula, Sporidiobolus, Sporobolomyces, Stetigmatomyces, Trichosporon

Activators:
SH compound such as cystein, 2-mercaptoethanol of dithiothreitol

Storage and Stability:
Stable for 1 year at 4ºC. About 90% of the lytic activity is lost when stored at 30ºC for 3 months.

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 brewer's 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 20ºC 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.