Induction of Sesquiterpene Cyclase During Integrated Extraction of Sesquiterpenes from Hairy Root Cultures of Hyoscyamus muticus

Hyoscyamus muticus의 모상근배양으로부터 Sesquiterpene 화합물의 Intergration 추출시 Sesquiterpene Cyclase의 유도

  • BACK, Kyoungwhan (Department of Agronomy, Kyungpook National University) ;
  • SHIN, Dong Hyun (Department of Agronomy, Kyungpook National University) ;
  • KIM, Kil Ung (Department of Agronomy, Kyungpook National University) ;
  • De HAAS, Cynthia R. (Department of Chemical Engineering, The Pennsylvania State University) ;
  • CHAPPELL, Joseph (Department of Agronomy, University of Kentucky) ;
  • CURTIS Wayne R. (Department of Chemical Engineering, The Pennsylvania State University)
  • Published : 1997.09.01

Abstract

The sesquiterpene cyclase (SC) was induced and its products were accumulated in the culture media of Hyoscyamus muticus hairy roots by addition of Rhizoctonia solani extracts. The cumulative production of solavetivone was nearly doubled by integrated extraction of the products from the media during the 24 h accumulation period. Western blots with monoclonal antibodies against SC show that the enzyme levels are the same for both extracted and non-extracted cultures. SC activities measured in vitro with radioactive substrate are not significantly different. These results suggest that productivity is controlled by substrate availability within the terpenoid pathway, and feedback regulation precedes the branch-point enzyme sesquiterpene cyclase.

Rhizoctonia solani 추출물(elicitor)을 Hyoscyamus muticus 모상근배양에 처리하였을 때 sesquiterpene cyclase (SC)가 유도됨과 동시에 sesquiterpene화합물이 합성되어 배양액속에 축적하는 것으로 나타났다. 24시간 배양기간 동안 배양액으로부터 sesquiterpene을 추출ㆍ분리하여 배양할 경우, 추출ㆍ분리하지 않은 모상근배양보다 거의 두배의 함량이 생성되었다. Cyclase monoclonal antibody을 이용하여 immunoblot을 시도한 결과, 추출한 모상근배양과 추출하지 많은 모상근배양에서의 SC 절대량은 동일한 것으로 나타났으며, 효소활성도 현저한 차이가 없는 것으로 측정되었다. 이러한 결과는 terpenoid pathway에서 sesquiterpene 생합성이 이용가능한 기질의 량에 의해 조절되는 것으로 사료되며, feedback 조절은 sesquiterpene cyclase 효소에 앞서 일어나는 것으로 추정된다.

Keywords

References

  1. App Microbiol Biotechnol v.30 Stimulation of ajmalicine production and excretion from Catharanthus roseus: effects of adsorption in situ, elicitors and alginate immobilization Asada M;Shuler ML
  2. Biochem J v.19 A note on the kinetics of enzyme action Briggs GE;Haldane JBS
  3. Tibtech v.11 Increasing secondary metabolite production in plant-cell culture by redirecting transport Brodelius P;Pederson P
  4. Phytochemistry v.29 Two phase culture for the enhanced production of benzophenanthridine alkaloids in cell suspensions of Eschscholtzia californica Byun SY;Pederson H;Chin CK
  5. Biotechnol Bioeng v.42 Enhanced recovery of solavetivone from hairy root cultures of Hyoscyamus muticus using integrated product extraction Corry JP;Reed WL;Curtis WR
  6. Methods Carbohydr Chem v.1 General color reactions Dische Z
  7. Enzymes Dixon M;Wedd EC
  8. Proc N Y Acad Sci v.655 Approaches to understanding and manipulating the biosynthetic potential of plant roots Flores HE;Curtis WR
  9. Plant Physiol v.94 Phenylpropanoid metabolism in suspension cultures of Vanilla planifolia Andr. II Effects of precursor feeding and metabolic inhibitors Funk C;Brodelius PE
  10. Z. Naturforsch v.35 Influence of medium composition in the formation of secondary compounds in cell suspension cultures of Catharanthus roseus Knobloch KH;Berlin J
  11. Appl Microbiol Biotechnol v.36 High-level production of arbutin from hydroquinone in suspension cultures of Catharanthus roseus plant cells Inomata S;Yokoyama M;Seto S;Yanagi M
  12. Biochem v.Z49 Die kinetic der invertinwirkung Michaelis L;Menten ML
  13. Regulation in metabolism, Chap 1 Newsholme EA;Start C
  14. Biotechnol Bioeng v.39 Akaloid production by plant cell cultures of Holarrhena antidysenterica II. Effect of precursor feeding and cultivation in stirred tank bioreactor Panda AK;Bisaria VS;Mishra S
  15. J Chromatography v.657 Reversed-phase liquid chromatographic isolation of lubimin and solavetivone from Hyoscyamus muticus hairy root cultures Reddy GR;Signs MW;Flores HE;Curtis WR
  16. Phytochemistry v.27 Biotransformation of 3-oxo-p-methane derivatives by cultured cells of Nicotiana tabacum Suga T;Hirata T;Hamada S
  17. Phytochemisty v.27 Co-ordinated inhibition of squalene synthetase and induction of enzymes of sesquiterpene phytoalexin biosynthesis in cultures of Nicotiana tabacum Threlfall DR;Whitehead IM
  18. Current Opin Biotechnol v.3 Integrated product formation and recovery in fermentation van der Wielen L;Luyben K
  19. Plant Physiol v.88 Induction of sesquiterpene cyclase and suppression of squalene synthetase activities in plant cell cultures treated with fungal elicitor Vogeli U;Chappell J
  20. Plant Physiol v.94 Regulation of a sesquiterpene cyclase in cellulase-treated tobacco cell suspension cultures Vogeli U;Chappell J