Westase (yeast cell wall lytic enzyme) | Enzymes for research

Westase(yeast cell wall lytic enzyme)

It is possible to dissolve yeast cell walls and prepare various types of DNA and intracellular enzymes.
Yeast protoplasts can be prepared.

Model	Sales destination	Sales Code No.	Eur-lex.europa.eu eur-lex.europa.eu
Westase	Takara Bio Inc.	9005	1 gr

Features	This is a complex enzyme preparation for lysing yeast cell walls, primarily consisting of β-1,6 glucanase and β-1,3 glucanase activity. In particular, its main activity is β-1,6 glucanase, which differentiates it from other enzyme preparations for lysing yeast cell walls. This product is effective not only for ascomycete yeasts but also for basidiomycete yeasts and imperfect yeasts, enabling protoplast preparation and achieving a high protoplast regeneration rate.Since no DNase activity is observed under recommended conditions, it can also be used to prepare DNA from yeast.
Shape	Lyophilized powder (contains diatomaceous earth as an excipient)
save	4℃, dry conditions
起源	Streptomyces rochei DB-34
Specification	β-1,6 glucanase activity (37℃) 400 U/g powder or more Cell wall lytic activity (30℃) 35,000 U/g powder or more
至適温度	30 ~ 50
Optimal pH	6.0
How To Use	Dissolve an appropriate amount in McIlvaine Buffer (0.1 M citric acid solution and 0.2 M disodium hydrogen phosphate solution in a ratio of approximately 36.8:63.2, adjusted to pH 6.0), and filter through a cellulose acetate filter before use. Sodium tartrate dihydrate must be used as an osmotic adjuster when preparing protoplasts. The target yeast must be in the logarithmic growth phase.

Protoplast formation for various yeasts

strain	Protoplast formation rate (%)
strain	Westase	Zymolyase
Schizosaccharomyces pombe OUT 0351	○	△
Saccharomyces cerevisiae X2180-1A	○	○
Zygosaccharomyces rouxii OUT 1130	×	○
Hansenula mrakii RIB 522	○	○
Kluyveromyces lactis IFO 0433	○	○
Anomalous pichia OUT 10213	○	○
Lipomyces starkeyi OUT 10381	○	△
Filobasidium floriforme OUT 1915	○	×
Ustilago maydis OUT 5346	○	×
Rhodosporidium toruloides OUT 10512	○	×
Tremella mesenterica OUT 9310	○	×
Graphiola phoenicis OUT 9100	○	×
Sporobolomyces roseus OUT 1105	×	×
Brettanomyces brusselsensis OUT 0797	○	○
Candida colliculosa OUT 0663	○	○
Candida tropicalis OUT 1400	△	○
Candida utilis OUT 0639	○	○
Kloeckera apiculata OUT 0865	○	○
Rhodotorula glutinis OUT 1125	×	×
Trigonopsis variabilis OUT 0755	○	○
Cryptococcus albidus OUT 0612	○	×
Phaffia rhodozyma OUT 10129	○	○

Protoplast formation rate: ○: 80% or more, △: 50% or more, ×: 0%

Yoshida, M., Nishi, A., Obuchi, K., Hojo, T., Matsuzawa, A., Hamaji, M., and Kumagai, C. (1997).
Biotechnology, 75, 229.
Nishi, A., Obuchi, K., Hamaji, M., and Kumagai, C. (1999).
Biotechnology, 77, 60.
Obuchi, K., Nishi, A., Hamaji, M., and Kumagai, C. (1999).
Biotechnology, 77, 137.

Is filtration required when using?: The diatomaceous earth contained as an excipient is insoluble, so without filtration, a precipitate will form and the product will be difficult to handle.
Please be sure to filter the water. Diatomaceous earth also acts as a filter aid.

Does the buffer have to be McIlvaine?: It doesn't necessarily have to be McIlvaine. Try a buffer that is easy to use depending on your purpose.

When I used sorbitol as an osmotic adjuster, I was unable to produce protoplasts. Why?: This enzyme cannot exert its full effect unless an organic acid salt is used as an osmotic pressure adjuster.
Be sure to use an organic acid salt, preferably sodium tartrate. When regenerating protoplasts for transformation, use sorbitol as an osmotic agent.

What is the appropriate concentration of enzyme to use?: A concentration of 0.5% during reaction is expected to be sufficient, but depending on the application, a lower concentration may be preferable.