Protoplast formation from schizophyllum commune by a culture filtrate of bacillus circulans KA-304 grown on a cell-wall preparation of S. Commune as a Carbon Source;

Abstract

From microorganisms growing on a cell-wall preparation (CWP) of Schizophyllum commune as a carbon source, Bacillus circulans KA-304 was isolated on the bases of activities in culture filtrate to decrease turbidity of the CWP-suspension and to form protoplasts from S. commune. The culture filtrate was also active in hydrolyzing p-nitrophenyl (p-NP)-α-d-glucoside, p-NP-β-d-glucoside, and p-ΝΡ-β-d-Ν-acetylglucosaminide. The protoplast-forming and the p-NP-glycoside-hydrolyzing activities were increased by the addition of CWP of S. commune to the culture medium, and this was not observed for other bacteria tested (15 genera, 80 species). B. circulans KA-304 was shown on gel filtration to produce at least two enzyme species for hydrolyzing both p-NP-α-d-glucoside and p-NP-α-d-glucoside. The protoplast-forming activity was retained for at least 6 months at 5°C as an ammonium sulfate (90% saturation) precipitate or at −20°C as a freeze-dried preparation (KA-preparation). The activity was stable at pH 6.5–7.0, and remained after 10 min of treatment at 40°C. Protoplast formation proceeded optimally at pH 6.5 with 50 mm potassium phosphate buffer and 0.5 m mannitol as an osmotic stabilizer. B. circulans KA-304 seems to be a suitable strain producing enzyme(s) to prepare protoplasts from S. commune. © 1997, Taylor & Francis Group, LLC. All rights reserved.