Zymolyase® from Arthrobacter luteus, min. 100 U/mglyoph.
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Zymolyase®, produced by a submerged culture of Arthrobacter
luteus (1), has strong lytic activity against living yeast cell walls to
produce protoplast or spheroplast of various strains of yeast cells (2, 3).
This enzyme is prepared by ammonium sulfate precipitation and further purified by affinity chromatography. The essential enzyme activity for the lysis of yeast cells 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). Lytic activity varies depending on yeast strain, growth stage of yeast, or cultural conditions (6, 7, 8).
Contained main side activities are β-1,3-glucanase, protease, and mannanase (3).
At 30 °C about 90 % of the lytic activity is lost after 3 months and at 60 °C after 5 minutes all the lytic activity is lost.
For lysis of viable cells the optimum temperature is 35 °C at pH 7.5 and for hydrolysis of yeast glucan 45 °C at pH 6.5.
Zymolyase® was shown to lyse Ashbya, Candida, Debaryomyces, Eremothecium, Endomyces, Hansenula, Hanseniaspora, Kloeckera, Kluyveromyces, Lipomyces, Metschikowia, Pichia, Pullularia, Torulopsis, Saccharomyces, Saccharomycopsis, Saccharomycodes, Schwanniomyces, etc. (5). It is activated by a SH compound such as cysteine, 2-mercaptoethanol or dithiothreitol.
Instructions for use:
Zymolyase® may not completely be dissolved in buffers and is therefore used as a suspension. A 2 to 10 % stock solution in 10 mM sodiumphosphate buffer (pH 7.4) or 50 mM Tris-Cl (pH 7.5), respectively, containing 5 % glucose and 50 % glycerol each may be prepared. The suspension may be stored in aliquots at -20 °C. The working concentration is 2 - 5 mg/ml and the suspension can be optionally sterilized by filtration (0.2 µm, no nitrocellulose filter). Zymolyase® may as well be solubilized in freshly prepared working buffer (e.g. 50 mM Tris-Cl, pH 7.5, 10 mM EDTA, 0.3 % 2-Mercaptoethanol) in the required working concentration directly prior to use.
Unit definition: One unit of lytic activity is defined as the enzyme amount causing a decrease of 30 % in absorbance at 800 nm using 6 mg Brewer's yeast as substrate in phosphate buffer (pH 7.5) at 25 °C.
Zymolyase = registered trademark of Kirin Holdings Company Limited
- Kaneko, T. et al. (1969) J. Gen. Appl. Microbiol. 15, 317 ff.
- Kitamura, K. et al. (1971) Arch. Biochem. Biophys. 145, 402 ff.
- Kitamura, K. et al. (1972) J. Gen. Appl. Microbiol. 18, 57 ff.
- Kitamura, K. & Yamamoto, Y. (1972) Arch. Biochem. Biophys. 153, 403 ff.
- Kaneko, T. et al. (1973) Agric. Biol. Chem. (1973) 37, 2295 ff.
- Kitamura, K. et al. (1974) J. Gen. Appl. Microbiol. 20, 323 ff.
- Kitamura, K. & Yamamoto, Y. (1981) Agric. Biol. Chem. 45, 1761 ff.
- Kitamura, K. & Tanabe, K. (1982) Agric. Biol. Chem. 46, 553 ff.
|Precautions||P261 - P284|
|Reaction||P304 +P340 - P342 +P311|
Storage Temperature: +2 °C to +8 °C