Molecular Breeding

, Volume 11, Issue 1, pp 59–67 | Cite as

Overproduction of an alkali- and thermo-stable xylanase in tobacco chloroplasts and efficient recovery of the enzyme

  • Sadhu Leelavathi
  • Naveen Gupta
  • Shankar Maiti
  • Amit Ghosh
  • Vanga Siva Reddy
Article

Abstract

Overproduction of cellulolytic enzymes through conventional nucleartransformation approaches posed a major challenge as they can potentiallydegrade the cell wall components and thereby affect transgenic plant growth anddevelopment. In this study, we have tested the possibility to over produce analkali-thermostable xylanase gene from Bacillus sp. StrainNG-27 in tobacco plants through chloroplast expression. Our results showed thatthe xylanase expression can reach up to 6% of the total soluble protein, avaluecomparable to high level expression reported for several non-cellulolyticproteins in tobacco chloroplasts. The chloroplast-expressed xylanase retainedits activity even when the leaves were dried under sun or at 42°C, offering flexibility in the agricultural system intransport and storage. The recombinant enzyme was purified to homogeneity usingsingle step chromatography with more than 85% recovery. Most importantly,transgenic plants were indistinguishable from the control untransformed plantsin their morphology, growth and in seed setting. These results open up newavenues for large scale production of several other industrially usefulcellulolytic enzymes through chloroplast expression.

Cellulolytic enzymes Chloroplast expression Food technology Paper pulp industry Protein purification 

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Copyright information

© Kluwer Academic Publishers 2003

Authors and Affiliations

  • Sadhu Leelavathi
    • 1
  • Naveen Gupta
    • 2
  • Shankar Maiti
    • 2
  • Amit Ghosh
    • 2
  • Vanga Siva Reddy
    • 1
  1. 1.International Center for Genetic Engineering & BiotechnologyNew DelhiIndia
  2. 2.Sector 39-AInstitute of Microbial TechnologyChandigarhIndia

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