Abstract:The effects of different factors, such as age, lysozyme dosage, enzymatic time, the pH value, temperature, osmotic stabilizer, and penicillin, on protoplast formation and regeneration of Bacillus strains DC-12 were studied. The most suitable hypertonic buffer, culture time, pH value, lysozyme dosage, reaction temperature and time were HM hypertonic buffer, 7.5, 2.0 mg/mL, 37 ℃ and 60 min, respectively, with which the rate of protoplast formation and regeneration were 95.8%. If 0.6 mol/L NaCl was used as the regeneration hypertonic buffer, the rate of protoplast formation and regeneration was 21.6%.
