Planta Med 2002; 68(4): 336-340
DOI: 10.1055/s-2002-26739
Original Paper
Biochemistry, Physiologie, in vitro cultures
© Georg Thieme Verlag Stuttgart · New York

Production of Paclitaxel and Baccatin III in a 20-L Airlift Bioreactor by a Cell Suspension of Taxus wallichiana

A. Navia-Osorio1 , H. Garden2 , R. M. Cusidó1 , J. Palazón1 , A. W. Alfermann2 , M. T. Piñol1
  • 1Laboratorio de Fisiologia Vegetal, Facultad de Farmacia. Universidad de Barcelona, Barcelona, Spain
  • 2Institut für Entwicklungs- und Molekularbiologie der Pflanzen, Heinrich-Heine-Universität Düsseldorf,
    Düsseldorf. Germany
Further Information

Publication History

May 28, 2001

August 5, 2001

Publication Date:
02 May 2002 (online)

Abstract

A cell suspension culture of Taxus wallichiana (Himalayan Yew) was grown in shake flasks and a 20-L airlift bioreactor running for 28 days in a batch mode, and its capacity to accumulate paclitaxel and baccatin III was measured. When both culture types were in the highest productive state (from day 24 to day 28), there was a greater accumulation of paclitaxel and baccatin III in the bioreactor culture than in the shake flask culture (factor of 2.0 and 1.2, respectively). These increases in paclitaxel and baccatin III production cannot be related to the difference observed between the growth rates of both cultures, because when the bioreactor culture was at maximum productivity, its cell biomass, expressed in g L-1 of dry weight, was similar to that obtained in the shake flask culture. It seems that these improvements were mainly due to adequate aeration and mixing of the culture in the bioreactor. The maximum yield observed for paclitaxel (20.84 mg L-1 day 24) and baccatin III (25.67 mg L-1 day 28) represents a productivity of 0.90 mg L-1 d-1 and 0.93 mg L-1 d-1 respectively.

References

  • 1 Fett-Neto A G, DiCosmo F. Distribution and amounts of taxol in different shoot parts of Taxus cuspidata .  Planta Medica. 1992;  58 464-6
  • 2 Guénard D, Guéritte-Voegelein F, Potier P. Taxol and taxotere: Discovery, chemistry and structure activity relationships.  Adv. Chem. Res.. 1993;  26 160-7
  • 3 Hezari M, Ketchum R EB, Gibson D M, Croteau R. Taxol production and taxadiene synthase activity in Taxus canadensis cell suspension cultures.  Arch. Biochem. Biophys.. 1997;  337 185-90
  • 4 Nicolaou K C, Yang Z, Liu J J, Ueno H, Nantermet P G, Guy R K, Clalborne C F, Renaud J, Couladouros E A, Paulvannan K, Sorensen E J. Total synthesis of taxol.  Nature. 1900;  367 630-4
  • 5 Fett-Neto A G, Zhang W Y, DiCosmo F. Kinetics of taxol production, growth and nutrient uptake in cell suspensions of Taxus cuspidata .  Biotechnol. Bioeng.. 1994;  44 205-10
  • 6 Kim D, Pedersen H M, Chin C K. Cultivation of Thalictrum rugosum cell suspension in an improved airlift bioreactor: Stimulatory effect of carbon dioxide and ethylene on alkaloid production.  Biotechnol. Bioeng.. 1991;  38 331-9
  • 7 Srinivasan V, Pestchanker L, Moser S, Hirasuna T, Taticek R A, Shuler M L. Taxol production in bioreactors: Kinetics of biomass accumulation, nutrient uptake, and taxol production by cell suspensions of Taxus baccata .  Biotechnol. Bioeng.. 1995;  47 666-76
  • 8 Cusidó R M, Palazón J, Navia-Osorio A, Mallol A, Bonfill M, Morales C, Piñol M T. Production of Taxol® and baccatin III by a selected Taxus baccata callus line and its derived cell suspension culture.  Plant Sci.. 1999;  146 101-7
  • 9 Vanêk T, Malá J, Šaman D, Šilhavá I. Production of taxanes in a bioreactor by Taxus baccata cells.  Planta Med.. 1999;  65 275-6
  • 10 Son S H, Choi S M, Lee Y H, Choi K B, Yun S R, Kim J K, Park H J, Kwon O W, Noh E W, Seon J H, Park Y G. Large-scale growth and taxane production in cell cultures of Taxus cuspidata (Japanese yew) using a novel bioreactor.  Plant Cell Reports. 2000;  19 628-33
  • 11 Gamborg O L, Miller R A, Ojima K. Nutrient requirements of suspension cultures of soybean root cells.  Exp. Cell. Res.. 1968;  50 151-8
  • 12 Fischer U, Santore U J, Hüsemann W, Barz W, Alfermann  A W. Semicontinuous cultivation of photoautotrophic cell suspension cultures in a 20 l airlift-reactor, Plant Cell.  Tissue and Organ Culture. 1994;  38 123-34
  • 13 Duncan D R, Widholm J M. Measurements of viability suitable for plant tissue cultures. In: Pollard JW, Walker JM, editors Methods in molecular Biology. Vol. 6 New Jersey; The Humana Press 1990: 29-37
  • 14 Grothaus P G, Bignami G S, O’Malley S, Harada K E, Byrnes J B, Waller D F, Raybould T JG, McGuire M T, Alvarado B. Taxane-specific monoclonal antibodies: Measurement of taxol, baccatin III, and ”total taxanes” in Taxus brevifolia extracts by enzyme immunoassay.  J. Nat. Prod.. 1995;  58 1003-14
  • 15 Guern J, Felle H, Mathien Y, Kurkdjian A. Regulation of intracellular pH in plant cells.  International Review of Cytology. 1991;  127 111-73
  • 16 Stepan-Sarkissian G. Selection of media for tissue and cell culture. In: Pollard JW, Walker JM, editors Methods in Molecular Biology. Vol. 6 New Jersey; The Humana Press 1990: 1-27
  • 17 Ketchum R E, Gibson D M, Croteau R B, Shuler M L. The kinetics of taxoid accumulation in cell suspension cultures of Taxus following elicitation with methyljasmonate.  Biotechnol. Bioeng.. 1999;  62 97-105
  • 18 Srinivasan V, Ciddi V, Bringi V, Shuler M L. Metabolic inhibitors, elicitors and precursors as tools for probing yield limitation in taxane production by Taxus chinensis cell cultures.  Biotechnol. Prog.. 1996;  12 457-65
  • 19 Hamada H, Sanada K, Furuya T, Kawabe S, Jaziri M. Biotransformation of paclitaxel by cell suspension cultures of Eucalyptus perrniana .  Natural products Letters. 1996;  9 47-52
  • 20 Scragg A H, Morris P, Allan E J, Bond P, Fowler M W. Effect of scale-up on serpentine formation by Catharanthus roseus suspension cultures.  Enzyme Microb. Technol.. 1987;  9 619-24
  • 21 Seki M, Ohzora C, Takeda M, Furusaki S. Taxol (Paclitaxel) production using free and immobilized cells of Taxus cuspidata .  Biotechnol. Bioeng.. 1997;  53 214-9

Prof. M. Teresa Piñol

Laboratorio de Fisiologia Vegetal

Facultad de Farmacia. Universidad de Barcelona


Avda. Diagonal 643

08028 Barcelona

Spain

Email: pinyol@farmacia.far.ub.es

Phone: +43-93 4024492

Fax: +43-93 4029043