Zymolyase®

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 cells^2,3 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 unit^4,5,10,11.

Benefits

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

Applications

Protoplast/spheroplast preparation
Yeast cell fusion
Transformation of yeast cells
Yeast genetics

Name	Datasheet	Packsize	Order
Zymolyase (R) 100T (Arthrobacter luteus)		500 mg (100 KU/g)	View
Zymolyase (R) 20T (Arthrobacter luteus)		1 g (20 KU/g)	View

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 below^12-16.

Supporting Documents

Specifications

Form	Lyophilized powder
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
Stable pH	5~10
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

Reconstitution

The best reconstitution buffer for Zymolyase is M/15 Phosphate buffer, pH 7.5 (which is also the reaction buffer given on the product datasheets). Citrate buffer or Hepes buffer can be also used. (You can also use PBS buffer for reconstitution of zymolyase 120491 in place of the recommended phosphate buffer if you keep same pH 7.5)

Precise ingredients / recipe used for the "M/15 phosphate buffer, pH 7.5" suggested on datasheet are as follows:

>> Na2HPO4・12H2O; 95.5g/4L(PW) + KH2PO4; 9.07g/L(PW)

PW: Pure Water

Note also: Use a sterilized filter (except nitrocellulose) when a sterilized enzyme solution is needed.

Storage of Reconstituted Enzyme

The enzyme can be stored for one year at -80 after reconstitution in Phosphate buffer, citrate buffer or Hepes buffer.

We suggest storing in single-use aliquots to minimise freeze thaw cycles.

Properties

Lytic Spectrum by Zymolyase

Susceptible strains in low concentration (0.2 units/ml)

Ashbya, Endomyces, Kloeckera, Kluyveromyces, Pullularia, Saccharomyces

Susceptible strains in high concentration (2.0 units/ml)

Candida, Debaryomyces, Eremothecium, Hansenula, Hanseniaspora, Lipomyces, Metschikowia, Saccharomycopsis, Saccharomycodes, Schizosaccahromyces, Selenozyma, Trigonopsis,Wickerhamia

Susceptibility depending on strains

Bretanomyces, Cryptococcus, Nadsonia, Pichia, Rodosporidium, Schwanniomyces, Stephnoascus, Torulopsis

No susceptible strains

Bullera, Pityrosporum, Rhosotorula, Sporidiobolus, Sporobolomyces, Stetigmatomyces, Trichosporon

Buffers tested with Zymolyase

The following phosphate buffers (pH 7.5) have been tested for Zymolyase: (Citrate buffer or Hepes buffer can be also used. You can also use PBS buffer for reconstitution of zymolyase 120491 in place of the recommended phosphate buffer if you keep same pH 7.5 as stated under “Reconstitution” above)

Buffer	Concentration	Yeast lytic activity (unit/g)
Potassium/Sodium Phosphate Buffer	1/15 M	21,000
Potassium Phosphate Buffer	5 mM	20,300
	10 mM	21,400
	50 mM	21,000
Sodium Phosphate Buffer	5 mM	20,500
	10 mM	21,200
	50 mM	21,200

Assay for Enzyme Activity

Unit definition

One unit of lytic activity is defined as that amount which indicates 30% of decrease in absorbance at 800 nm (A800) of the reaction mixture under the following condition.

Reaction mixture

Enzyme Solution	1ml - 0.05-0.1 mg/ml for Zymolyase® - 20T 1ml - 0.012-0.024 mg/ml for Zymolyase® - 100T
Substrate	3ml - Brewer's yeast cell suspension (2 mg dry weight/ml)
Buffer	5ml - M/15 Phosphate buffer, pH 7.5
Distilled Water	1ml

Preparation of M/15 phosphate buffer

Dissolve 19.1 g of di-sodium hydrogenphosphate 12-hydrate in distilled water to make an 800 mL solution
Dissolve 1.814 g of potassium dihydorgenphosphate in distilled water to make a 200 mL solution
Mix the two solutions together.

Citations

Recent Citations

Domestication and divergence of Saccharomyces cerevisiae beer yeasts.
Gallone, B., et al (2016) Cell, 166(6), 1397-1410.

Keeping Candida commensal: how lactobacilli antagonize pathogenicity of Candida albicans in an in vitro gut model
Graf, K., Last, A., Gratz, R., Allert, S., Linde, S., Westermann, M., ... & Hube, B. (2019). Disease models & mechanisms, 12(9), dmm039719.

Structure-based directed evolution improves S. cerevisiae growth on xylose by influencing in vivo enzyme performance.
Lee, M., Rozeboom, H. J., Keuning, E., de Waal, P., & Janssen, D. B. (2020). Biotechnology for Biofuels, 13(1), 5.

Crosstalk between chromatin structure, cohesin activity and transcription.
Maya-Miles, D., Andújar, E., Pérez-Alegre, M., Murillo-Pineda, M., Barrientos-Moreno, M., Cabello-Lobato, M. J., ... & Prado, F. (2019). Epigenetics & chromatin, 12(1), 47.

Molecular Wiring of a Mitochondrial Translational Feedback Loop.
Salvatori, R., Kehrein, K., Singh, A. P., Aftab, W., Möller-Hergt, B. V., Forne, I., ... & Ott, M. (2019). Molecular Cell.

The Aspartic Protease Ddi1 Contributes to DNA-Protein Crosslink Repair in Yeast.
Serbyn, N., Noireterre, A., Bagdiul, I., Plank, M., Michel, A. H., Loewith, R., ... & Stutz, F. (2020). Molecular Cell.

Zymolyase 100T- highly cited

Tor-mediated induction of autophagy via an Apg1 protein kinase complex.
Kamada, Y., et al (2000). The Journal of Cell Biology, 147(2), 435-446.

Formation process of autophagosome is traced with Apg8/Aut7p in yeast.
Kirisako, T., et al (1999). The Journal of Cell Biology, 147(2), 435-446.

Nuclear localization of Cdc25 is regulated by DNA damage and a 14-3-3 protein.
Lopez-Girona, A., et al (1999). Nature, 397(6715), 172-175.

Aggregation of huntingtin in yeast varies with the length of the polyglutamine expansion and the expression of chaperone proteins.
Krobitsch, S., & Lindquist, S. (2000). PNAS, 97(4), 1589-1594.

Zymolyase 20T- highly cited

Construction and characterization of a yeast artificial chromosome library containing seven haploid human genome equivalents.
Albertsen, H.M., et al (1990). PNAS, 87(11), 4256-4260.

Sequential assembly of myosin II, an IQGAP-like protein, and filamentous actin to a ring structure involved in budding yeast cytokinesis.
Lippincott, J., & Li, R. (1998). The Journal of Cell Biology, 140(2), 355-366.

ATM activation and its recruitment to damaged DNA require binding to the C terminus of Nbs1.
You, Z., et al (2005). Molecular and Cellular Biology, 25(13), 5363-5379.

SAC1-like domains of yeast SAC1, INP52, and INP53 and of human synaptojanin encode polyphosphoinositide phosphatases.
Guo, S., et al (1999). JBC, 274(19), 12990-12995.

Additional Resources

Zymolyase

Type: Flyers

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We are a global provider of human and animal biospecimens: including frozen & FFPE tissue, DNA, RNA, total proteins, blood products and primary cells.

We are a global provider of human and animal biospecimens: including frozen & FFPE tissue, DNA, RNA, total proteins, blood products and primary cells.

We are a global provider of human and animal biospecimens: including frozen & FFPE tissue, DNA, RNA, total proteins, blood products and primary cells.

We are a global provider of human and animal biospecimens: including frozen & FFPE tissue, DNA, RNA, total proteins, blood products and primary cells.

We are a global provider of human and animal biospecimens: including frozen & FFPE tissue, DNA, RNA, total proteins, blood products and primary cells.

We are a global provider of human and animal biospecimens: including frozen & FFPE tissue, DNA, RNA, total proteins, blood products and primary cells.

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The CELLBANKER® series of cryopreservation media allows for the stable long-term storage of any cell type, including sensitive cell lines.

The CELLBANKER® series of cryopreservation media allows for the stable long-term storage of any cell type, including sensitive cell lines.

The CELLBANKER® series of cryopreservation media allows for the stable long-term storage of any cell type, including sensitive cell lines.

The CELLBANKER® series of cryopreservation media allows for the stable long-term storage of any cell type, including sensitive cell lines.

The CELLBANKER® series of cryopreservation media allows for the stable long-term storage of any cell type, including sensitive cell lines.

The CELLBANKER® series of cryopreservation media allows for the stable long-term storage of any cell type, including sensitive cell lines.

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Prepared from Arthrobacter luteus, Zymolyase has a highly effective lytic activity against yeast cell walls.

Prepared from Arthrobacter luteus, Zymolyase has a highly effective lytic activity against yeast cell walls.

Prepared from Arthrobacter luteus, Zymolyase has a highly effective lytic activity against yeast cell walls.

Prepared from Arthrobacter luteus, Zymolyase has a highly effective lytic activity against yeast cell walls.

Prepared from Arthrobacter luteus, Zymolyase has a highly effective lytic activity against yeast cell walls.

AMSBIO offers one of the largest selection of active recombinant enzymes and enzyme activity assays for profiling, inhibitor screening assays, and drug discovery.

AMSBIO offers one of the largest selection of active recombinant enzymes and enzyme activity assays for profiling, inhibitor screening assays, and drug discovery.

AMSBIO offers one of the largest selection of active recombinant enzymes and enzyme activity assays for profiling, inhibitor screening assays, and drug discovery.

AMSBIO offers one of the largest selection of active recombinant enzymes and enzyme activity assays for profiling, inhibitor screening assays, and drug discovery.

AMSBIO offers one of the largest selection of active recombinant enzymes and enzyme activity assays for profiling, inhibitor screening assays, and drug discovery.

AMSBIO offers one of the largest selection of active recombinant enzymes and enzyme activity assays for profiling, inhibitor screening assays, and drug discovery.

AMSBIO offers one of the largest selection of active recombinant enzymes and enzyme activity assays for profiling, inhibitor screening assays, and drug discovery.

AMSBIO offers one of the largest selection of active recombinant enzymes and enzyme activity assays for profiling, inhibitor screening assays, and drug discovery.

AMSBIO offers one of the largest selection of active recombinant enzymes and enzyme activity assays for profiling, inhibitor screening assays, and drug discovery.

AMSBIO offers one of the largest selection of active recombinant enzymes and enzyme activity assays for profiling, inhibitor screening assays, and drug discovery.

AMSBIO offers one of the largest selection of active recombinant enzymes and enzyme activity assays for profiling, inhibitor screening assays, and drug discovery.

AMSBIO offers one of the largest selection of active recombinant enzymes and enzyme activity assays for profiling, inhibitor screening assays, and drug discovery.

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We supply a range of superior magnetic products each uniquely suited to specific protein purification, proteomics and genomics applications.

Zymolyase®

Efficient degradation of yeast cell walls

Benefits

Applications

Supporting Documents

Specifications

Reconstitution

Storage of Reconstituted Enzyme

Properties

Lytic Spectrum by Zymolyase

Susceptible strains in low concentration (0.2 units/ml)

Susceptible strains in high concentration (2.0 units/ml)

Susceptibility depending on strains

No susceptible strains

Buffers tested with Zymolyase

Assay for Enzyme Activity

Unit definition

Reaction mixture

Preparation of M/15 phosphate buffer

Citations