1887

Abstract

Novozym 234, a commercially available enzyme from , has been used to prepare protoplasts from QM 9414. Optimal conditions were : 20 h old mycelium, 0·9 -KCl as the osmotic stabilizer. 0·1% (w/v) Novozym 234 and 18 h incubation at 28°C. To prevent damage to the protoplasts during isolation the incubation mixtures were agitated by vibration, so avoiding shaking or stirring. More than 95% of the protoplasts were viable and were also metabolically intact as shown by their ability to take up [C]leucine and incorporate it into cellular protein. Freshly prepared protoplasts could be induced by sophorose to produce and secrete carboxymethylcellulase and -glucosidase. Optimal conditions were: 0·9 -KCl as osmotic stabilizer, phosphate buffer pH 6·0, 7 m-sophorose and 10 protoplasts ml. Separation of the secreted proteins by fast protein liquid chromatography revealed at least seven peaks of carboxymethylcellulase activity and -glucosidase activity, indicating that some multiplicity of the forms of these enzymes is already present at the secretion stage.

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1985-06-01
2024-12-12
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