Optimization of Aconitine Production in Suspension Cell Cultures of Aconitum napellus L.

  • Hwang, Sung-Jin (Dept. of Oriental Medicine Materials & Biotechnolgy Industrialization Center (BIC), Dongshin Univ.) ;
  • Kim, Yun-Hee (Dept. of Oriental Medicine Materials & Biotechnolgy Industrialization Center (BIC), Dongshin Univ.) ;
  • Pyo, Byoung-Sik (Dept. of Oriental Medicine Materials & Biotechnolgy Industrialization Center (BIC), Dongshin Univ.)
  • Published : 2004.11.01

Abstract

Aconitine alkaloids produced from cell suspension cultures of Aconitum napellus for the first time. The effects of various culture conditions on cell biomass and aconitine accumulation in cell suspension cultures were investigated. Suspension cell cultures of A. napellus were established by transferring callus tissues from leaf explants onto liquid MS medium supplemented with $1\;mg/l$ NAA and $0.1\;mg/l$ kinetin. Among the culture media tested, MS medium had a pronounced effect on cell growth and aconitine accumulation. The maximum dry cell weight was obtained at inoculum size of 3 g (FCW) per flask and in MS medium supplemented with 5% sucrose after 8 weeks. The addition of salicylic acid (SA) and yeast extract (YE) in the MS medium enhanced aconitine accumulation. Using a proper combination of culture condition and supplements, aconitine content could reach 0.043% (dry weight basis), that was $2.5{\sim}3$ fold higher that detected in control cultures.

Keywords

References

  1. Cotin A, Heijden R, Hoopen HJG, Verpoorte R (1998) The inoculum size triggers typtamine or secologanin biosynthsis in Catharanthus roseous cell culture. Plant Sci. 139:205-211 https://doi.org/10.1016/S0168-9452(98)00188-5
  2. Dang X, Liu X, Feng J, Zhu M, Li A (1986) Analgesic action and no physical dependence of 3-acetylaconitine. Acta Pharm Sin. 7:413-418
  3. Draper J (1997) Salicyliate x superoxide synthesis and cell suicide in plant defense. Trends Plant Sci. 2:1360-1385
  4. Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res. 50:151-158 https://doi.org/10.1016/0014-4827(68)90403-5
  5. Gergorlo GH, Victor MLV (1997) Effects of acetylsalicylic acid on secondary metabolism of Catharanthus roseous tumor suspension cultures. Plant Cell Rep. 16:287-290
  6. Hikino H, Yamada TIC, Sato H, Konno C, Ohizumi Y (1979) Analgesic principles of Aconitum roots. J. Pharmocobiodyn. 2:78-83 https://doi.org/10.1248/bpb1978.2.78
  7. Hikino H, Konno C, Takata H, Yamada Y, Yamada C, Ohizumi Y, Sigio K, Fujimura H (1980) Antiinflammatory principles in acute inflammations. Eur. J. Pharmocobiodyn. 3:514-525 https://doi.org/10.1248/bpb1978.3.514
  8. Knobloch KH, Berlin J (1980) Influence of medium composition on the formation of secondary compounds in cell suspension cultures of Catharanthus roseous L. G. Don. Z. Naturforsch 35:551-556
  9. Liu JH, Zhu YX, Tang XC (1987) Anti-inflammatory and analgesic activities of N-deacetyllappsconitine and lappaconitine. Acta Pharm Sin. 8:301-305
  10. Lloyd GB, McCown BH (1980) Commercially feasible micropropagation of mountain laurel, Kalmia lati[olia, by the use of shoot top culture. Comb. Proc. Int. Plant Prop. Soc. 30:421-437
  11. Mehmetoglu WR, Curtis WR (1997) Effects of abiotic inducers on sequiterpene synthesis in hairy root and cell suspension cultures of Hyoscyamus muticus. App Biochem Biotechnol, 67:71-77 https://doi.org/10.1007/BF02787842
  12. Misawa M (1985) Production of useful plant metabolites. In Fiechter A Ed. Adv. Biochem. Eng. Biotechnol, Berlin Springer-Verlag pp. 59-88
  13. Morl T, Bando H, Kanaiwa Y, Wada K, Amiya T (1998) Studies on the constituents on the aconitine type alkaloids in decoction of kampo prescriptions containing Aconitum tuber. Natural Medicine 52:433-439
  14. Murasruge T, Skoog FA (1962) A revised medium for rapid growth and biosynthesis with tobacco tissue cultures. Physiol. Plant. 15:473-497 https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  15. Murayama M, Mori T, Banda H, Amiya T (1991) Studies on the constituents of Aconitum species. IX. The pharmacological properties of pyro-type aconitine alkaloids, components of processed aconite powder 'Kako-bushi-matsu' : analgesic, antiinflammatory and toxic activities. J. Ethnopharmac. 35:159-164 https://doi.org/10.1016/0378-8741(91)90068-O
  16. Ravishankar GA, Venkataraman LV (1993) Role of plant cell culture in food biotechnology: current trends, limitations and future prospects, In Prakash J, Pierik RLM, Eds. Plant Biotechnology : commercial prospects and problems. New Delhi, Oxford IBH Press p. 381-383
  17. Sakamoto K, Iida K, Sawamura K, Hajiro K, Asada Y, Yoshikawa T, Furuya T (1993) Effects of nutrients on anthocyanin production in cultured cells of Aralia cordata, Phytochemicals 33:357-360 https://doi.org/10.1016/0031-9422(93)85517-U
  18. Schenk RV, Hildrbrandt AC (1972), Medium and techniques for induction and growth of monocotyledonous and dicotyledonous plant cell cultures. Can J. Bot, 50:199-204 https://doi.org/10.1139/b72-026
  19. Su WW, Lei F (1993) Rosemarinic acid production in perfused Anchusa officinalis cultures: Effects of inoculum size. Biotechnol Lett, 15:1035-1038 https://doi.org/10.1007/BF00129933
  20. van Gulik WM, Nuutila AM, Vinke KL, ten Hoopen HJG, Heijnen JJ (1994) Effects of carbon dioxide, air flow rate, and inoculum density on the batch growth of Catharanthus roseous cell suspension in stirred ferments. Biotechnol, Prog. 10:335-339 https://doi.org/10.1021/bp00027a015
  21. Zhong JJ, Yoshida T (1995) High-density cultivation of Perilla frutescens cell suspensions for anthocyanin production : Effects of sucrose concentration and inoculum size, Enzyme Microb Technol, 17:1073-1079 https://doi.org/10.1016/0141-0229(95)00033-X