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Zymolyase® Yeast Cell Extraction
Zymolyase®, produced by a submerged culture of Arthrobacter luteus(1), has strong lytic activity against living yeast cell walls(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 with β-1,3-linkages and releases specifically laminaripentaose as the main and minimum product unit(4), (5), (10), (11).
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Product Info:
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 chromatography(9). Lytic activity varies depending on yeast strain, growth stage of yeast, or cultural conditions(6-8). Further informations related to Zymolyase® can be obtained in the reference section below(12-16).
Specification
| Product Name | Zymolyase®-20T | Zymolyase®-100T | |
|---|---|---|---|
| Form | Lyophilized Powder | ||
| Purification | Ammonium Sulfate Precipitation | Affinity Chromatography | |
| Activity | 20,000 units/g | 100,000 unites/g | |
| Essential enzyme | β-1,3-glucan laminaripentaohydrolase | ||
| Other activities contained (※1) |
β-1,3-glucanase | approx. 1.5 x 106 units/g | approx. 1.0 x 107 units/g |
| protease | approx. 1.0 x 104 units/g | approx. 1.7 x 104 units/g | |
| mannanase | approx. 1.0 x 106 units/g | approx. 6.0 x 104 units/g | |
| Contaminants | Amylase, Xylanase, Phosphatase |
Trace amount | Non detectable |
| Optimum pH and Temp. | pH7.5, 35°C (for lysis of viable yeast cells) pH6.5, 45°C (for hydrolysis of yeast glucan) |
||
| Stability | 2°C | No loss of activity was found after storage for 1 year | |
| Heat stability | 30°C | 70% of the lytic activity is lost after storage for 3 months | |
| 90% of the lytic activity is lost after storage for 3 months | |||
| 60°C | Lytic activity is lost on incubation for 5 minutes | ||
| Specificity (Lytic Spectrum) | Ashbya, Candida, Debaryomyces, Eremothecium, Endomyces, Hansenula, Hanseniaspora, Kloeckera, kluyveromyces, Lipomyces, Metschnikowia, Pichia, Pullularia, Torulopsis, Saccharomyces, Saccharomycopsis, Saccharomycodes, Schwanniomyces, etc. | ||
(※1) See reference, Kitamura, K., Kaneko, T., Yamamoto, Y., J. Gen. Appl. Microbiol., 18, 57 (1972) as to the definition of each enzyme units.
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 | 1 ml (0.05-0.1 mg/ml for Zymolyase®-20T) 1 ml (0.012-0.024 mg/ml for Zymolyase®-100T) |
|
| Brewer’s yeast cell suspension | 3 ml (2 mg/ml) | |
| 1/15M Phosphate buffer | 5 ml (pH7.5) | |
| Distilled water | 1 ml | |
After incubation for 2 hours at 25°C with gentle shaking, A800 of the mixture is determined. When 60% of A800 decrease, equivalent to 2 units, is observed in the reaction system, the brewer’s yeast cells are completely lysed, namely 1 unit of Zymolyase® lyses 3 mg dry weight of brewer’s yeast.
Reference
- Kaneko, T., Kitamura, K and Yamamoto, Y.: J. Gen. Appl. Microbiol., 15, 317 (1969)
- Kitamura, K., Kaneko, T. and Yamamoto, Y.: Arch. Biochem. Biophys., 145, 402 (1971)
- Kitamura, K., Kaneko, T. and Yamamoto, Y.: J. Hen. Appl. Microbiol., 18, 57 (1972)
- Kitamura, K. and Yamamoto, Y.: Arch. Biochem. Biophys., 153, 403 (1972)
- Kaneko, T., Kitamura, K. and Yamamoto, Y.: Agric. Biol. Chem., 37, 2295 (1973)
- Kitamura, K., Kaneko, T. and Yamamoto, Y.: J. Gen Appl. Microbiol., 20, 323 (1974)
- Kitamura, K. and Yamamoto,.: Agric. Biol. Chem., 45, 1761 (1981)
- Katamura, K. and Tanabe, K.: Agric, Biol. Chem., 46, 553 (1982)
- Katamura, K.: J. Ferment. Technol., 60, 257 (1982)
- Kitamura, K.: Agric. Biol. Chem., 46, 963 (1982)
- Kitamura, K.: Agric. Biol. Chem., 46, 2093 (1982)
- Calza R. E., Schroeder A. L.: J. Gen. Microbiol., 129, 413 (1983)
- Iizuka Masaru, Torii Yasuhiko, Yamamoto Takehiko: Agric. Biol. Chem., 47 (12), 2267 (1983)
- Shibata Nobuyuki, Kobayashi Hidemitsu, tojo Menehiro, Suzuki Shigeo: Arch. Biochem. Biophys., 251(2), 697 (1986)
- Iijima Y., Yanagi S. O.: Agric. Biol. Chem., 50 (7), 1855 (1986)
- Herrero Enrique, Sanz Pascual. Sentandreu Rafael: J. Gen. Microbiol., 133 (10), 2895 (1987)
Disclaimer: Biozatix Indonesia adalah distributor yang menyediakan alat- alat laboratorium, alat kesehatan, serta reagensia ELISA kits, immunohistochemistry (IHC), antibody, reagensia deteksi immunohistochemistry, secondary antibody untuk ELISA dan immunohistochemistry, research grade chemical, western blot, HPLC colomn, alat patologi, HPV genotyping, Florescent in Situ Hybridization (FISH), alat core biopsy.