Journal of Biosciences

, Volume 39, Issue 1, pp 33–41 | Cite as

Metabolic engineering of chloroplasts for artemisinic acid biosynthesis and impact on plant growth

  • Bhawna Saxena
  • Mayavan Subramaniyan
  • Karan Malhotra
  • Neel Sarovar Bhavesh
  • Shobha Devi Potlakayala
  • Shashi Kumar
Brief communication

Abstract

Chloroplasts offer high-level transgene expression and transgene containment due to maternal inheritance, and are ideal hosts for biopharmaceutical biosynthesis via multigene engineering. To exploit these advantages, we have expressed 12 enzymes in chloroplasts for the biosynthesis of artemisinic acid (precursor of artemisinin, antimalarial drug) in an alternative plant system. Integration of transgenes into the tobacco chloroplast genome via homologous recombination was confirmed by molecular analysis, and biosynthesis of artemisinic acid in plant leaf tissues was detected with the help of 13C NMR and ESI-mass spectrometry. The excess metabolic flux of isopentenyl pyrophosphate generated by an engineered mevalonate pathway was diverted for the biosynthesis of artemisinic acid. However, expression of megatransgenes impacted the growth of the transplastomic plantlets. By combining two exogenous pathways, artemisinic acid was produced in transplastomic plants, which can be improved further using better metabolic engineering strategies for commercially viable yield of desirable isoprenoid products.

Keywords

Antimalarial drug artemisinin gene pyramiding isopentenyl-diphosphate/isopentenyl pyrophosphate (IPP) metabolic flux mevalonate (MEV) pathway plastome transplastomics 

Notes

Acknowledgements

We thank the Department of Biotechnology (DBT) and Department of Science and Technology, India, for providing financial support to SK (grants BT/HRD/35/02/09/2008, BT/PR13028/PID/06/473/2009 and SR/SO/BB-37/2010 respectively). We also like to thank DBT for the grant to NSB for the 500 MHz NMR spectrometers at the ICGEB, New Delhi. Our sincere thanks to Dr Ranjan Nanda (ICGEB, New Delhi) for his help with the analysis of ESI-mass spectrometry data.

Supplementary material

12038_2013_9402_MOESM1_ESM.pdf (69 kb)
ESM 1(PDF 69.2 kb)

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Copyright information

© Indian Academy of Sciences 2014

Authors and Affiliations

  • Bhawna Saxena
    • 1
  • Mayavan Subramaniyan
    • 1
  • Karan Malhotra
    • 1
  • Neel Sarovar Bhavesh
    • 1
  • Shobha Devi Potlakayala
    • 2
  • Shashi Kumar
    • 1
  1. 1.International Centre for Genetic Engineering and BiotechnologyNew DelhiIndia
  2. 2.School of Science Engineering and TechnologyPenn State HarrisburgMiddletownUSA

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