creators_name: Leelavathi, Sadhu
creators_name: Gupta, Naveen
creators_name: Maiti, Shankar
creators_name: Ghosh, Amit
creators_name: Siva Reddy, Vanga
type: article
datestamp: 2012-02-06 18:17:57
lastmod: 2012-02-06 18:17:57
metadata_visibility: show
title: Overproduction of an alkali- and thermo-stable xylanase in tobacco chloroplasts and efficient recovery of the enzyme
ispublished: pub
subjects: QR
full_text_status: restricted
keywords: Cellulolytic enzymes, Chloroplast expression, Food technology, Paper pulp industry, Protein purification
note: Copyright of this article belongs to Springer Science.
abstract: Overproduction of cellulolytic enzymes through conventional nuclear transformation approaches posed a major
challenge as they can potentially degrade the cell wall components and thereby affect transgenic plant growth
and development. In this study, we have tested the possibility to over produce an alkali-thermostable xylanase
gene from Bacillus sp. Strain NG-27 in tobacco plants through chloroplast expression. Our results showed that
the xylanase expression can reach up to 6% of the total soluble protein, a value comparable to high level expression
reported for several non-cellulolytic proteins in tobacco chloroplasts. The chloroplast-expressed xylanase
retained its activity even when the leaves were dried under sun or at 42 °C, offering flexibility in the
agricultural system in transport and storage. The recombinant enzyme was purified to homogeneity using single
step chromatography with more than 85% recovery. Most importantly, transgenic plants were indistinguishable
from the control untransformed plants in their morphology, growth and in seed setting. These results open up
new avenues for large scale production of several other industrially useful cellulolytic enzymes through chloroplast
expression.
date: 2003
date_type: published
publication: Molecular Breeding
volume: 11
number: 1
publisher: Springer Science
pagerange: 59-67
id_number: doi:10.1023/A:1022168321380
refereed: TRUE
issn: 13803743
official_url: http://dx.doi.org/10.1023/A:1022168321380
related_url_url: http://www.springerlink.com/content/n35jgh03214vul13/
related_url_type: pub
citation:   Leelavathi, Sadhu and Gupta, Naveen and Maiti, Shankar and Ghosh, Amit and Siva Reddy, Vanga  (2003) Overproduction of an alkali- and thermo-stable xylanase in tobacco chloroplasts and efficient recovery of the enzyme.  Molecular Breeding, 11 (1).  pp. 59-67.  ISSN 13803743     
document_url: http://crdd.osdd.net/open/897/1/ghosh2003.pdf