Engineering Tolerance And Hyper Accumulation Of Heavy Metals In Recombinant E.coli Cells By Expressing PCS Gene From Leucaena Leucocephala | 4830
Journal of Biotechnology & Biomaterials
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Leucaena leucocephala, a leguminous tree with high biomass, has metal tolerance and survival ability in arid, infertile and
metal-contaminated areas. But so far, no reports are available on identification and functional characterization of the metaltolerance
genes in this species. In the present study, we report the isolation and characterization of phytochelatin synthase (LlPCS)
gene from this tree species. The predicted 3D structure of this protein exhibited potential possible binding sites for the metals
like Cd, Zn, Cu, Fe and Mn. The role of LlPCS gene in metal tolerance and accumulation was characterized in E. coli. The LlPCS
gene was sub-cloned into pET32a expression vector and bacterial transformation was performed. SDS-PAGE analysis of the total
protein isolated from the recombinant E. coli revealed the over expression of LlPCS protein along with His-tag. When grown in
toxic metal enriched media, recombinant E. coli exhibited enhanced tolerance to broad range of metals like arsenic, cadmium,
cobalt, copper, mercury, tin and zinc compared to controls. Further, these recombinant E. coli cells could also accumulate several
folds increased concentrations of toxic metals like arsenite, copper and arsenate compared to controls. Thus the present study
suggests that LlPCS gene confers enhanced toxic metal tolerance and accumulation in E. coli and might play crucial role in toxic
metal detoxification in Leucaena leucocephala. This study can help us in better understanding the toxic metal tolerance ability of
L. leucocephala for its successful usage in bioremediation technologies.
Merla Sri Lakshmi Sunita has completed her PhD in 2010 at the age of 29 years from Department of Genetics, Osmania University, Hyderabad. She
was awarded Dr. D.S. Kothari Postdoctoral fellowship from University Grants Commission, New Delhi and pursuing postdoctoral studies at Centre for
Biotechnology, Jawaharlal Nehru Technological University Hyderabad. She has published six papers in reputed national and international journals.
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