| Title | 고온성 clostridium thermocellum과 clostridium thermohydrosulfuricum의 원형질체 형성 및 재생 | ||
| Author | 김욱한 · 정기택 * · 이용현 | ||
| Address | 경북대학교 자연과학대학 유전공학과; * 농과대학 식품공학과 | ||
| Bibliography | Korean Journal of Microbiology, 28(4),304-310, 1990 | ||
| DOI | |||
| Key Words | Clostridium thermocellum, Clostridium thermohydrosulfuricum, protoplast formation, protoplast regeneration | ||
| Abstract | The conditions for protoplasts formation and regeneration of thermophilic anaerobic C. thermocellum and C. thermohydrosulfuricum were determined under the anaerobic growth conditions. The cells of C. thermocellum in initial exponential growth phase were identified to be the most suited for protoplast formation. The optimal conditions for protoplast formation were found to be at 37℃ for 2 hours with 0.5 mg/ml of lysozyme in TMG buffer (pH7.5). On the other hand, C. thermohydro-sulfuricum grown in the same medium but excluding glycine was optimally protoplasted at the same conditions but with 0.2 mg/ml of lysozyme. The protoplasts of both strains only subjected to lysozyme treatment of the short time were satisfactorily regenerated after 7-10 days incubation at 60℃ in regeneration medium containing 0.3-0.4 M sorbitol, 0.5% casamino acid, and high concentration of CaCl_2 and MgCl_2. The regeneration frequencies of the protoplasts of C. thermocellum and C. thermohydrosulfuricum were found to be very low level of 4.85×10^-3 and 4.23×10^-2, respectively. The nonregenerated L-form cells were also observed in regeneration medium together with regenerated cells. | ||
| Download PDF | Kor_280404_304-310p.pdf | ||