Selection of Protoplasts-Derived Cell Lines for High Yields Indole Alkaloids from Suspension Cultures of Vinca (Catharanthus roseus)

일일초(Catharanthus roseus) 현탁배양으로부터 원형질체유래 Indole Alkaloid 고생산성 세포주 선발

  • 김석원 (한국과학기술연구원 유전공학연구소 식품세포생물학연구실) ;
  • 정경희 (한국과학기술연구원 유전공학연구소 식품세포생물학연구실 한국과학기술연구원 유전공학연구소 식품세포생물학연구실 경희대학교 문리과대학 생물학과 서울대학교 공과대학 공업화학과 한국과학기술연구원 유전공학연구소 식품세포생물학연구실)
  • Published : 1991.04.01

Abstract

To produce economically important indole alkaloids by cell cultures, we selected protoplastsderived clones (protoclones) of vinca (Catharanthus roseus) for high yields of catharanthine and ajmalicine. Protoplasts were enzymatically isolated from suspension-cultured cells. The highest plating efficiency (1%) was obtained when protoplasts were plated at a density of 1$\times$105 protoplasts/ml in a culture medium solidified with 0.4% Seaplaque agarose. The growth rates of 40 protoclones subcultured on a solid medium varied over a wide range. Protoclone VPC-6, which had the highest growth rate, was observed to produce relatively high yields of catharanthine and ajmalicine when cultured in a liquid medium. Although the original cell line did not produce catharanthine at a detectable level by HPLC, protoclone VPC-10 produced it at a level of 5.9$\mu\textrm{g}$/g fresh weight of cells for 10 days of culture. Under the same conditions, protoclone VPC-15 produced ajmalicine at a level of 133.6$\mu\textrm{g}$/g, of which productivity was improved about ,3 times than that of the original cell line. The results indicate that differences in the growth rate and indole alkaloid yield among the protoclones reflect the somaclonal variation in suspnsion-cultured cells.

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