Journal of Applied Biosciences (J. Appl. Biosci.) [ISSN 1997 - 5902]
Volume 36: 2403 - 2408. Published December 7, 2010.
Characterization studies of thermostable alkaline phosphatase from various plant seeds
Vemuri Praveen Kumar*, Sunkara Prasanthi, Arumalla Chandra Sekhara Reddy, Nelapati Dhana Raj, Lakkoju Anudeep
Department of Biotechnology, K L University, Green Fields, Vaddeswaram, Guntur District, Andhra Pradesh, India
*Corresponding author: vemuripraveen@gmail.com
ABSTRACT
Objective: The objective of this study is to obtain and characterize thermostable alkaline phosphatase from plant seeds than produced prior from bacterial and animal sources.
Methodology and Results: Alkaline phosphatase was isolated and purified from plant seedlings by a streamline method without the use of proteolytic and lipolytic enzymes, butanol, or heavy metals. The study features are the use of a mild nonionic detergent, precipitated with acetone, maintained at low temperature to remove impurity, and the use of ion exchange columns to concentrate the alkaline phosphatase. The enzyme preparation revealed a major phosphatase band of 72 kDa and two minor activity bands. A variety of stability and activity profiles were determined for the purified seedling alkaline phosphatase: pH, temperature, surfactant and effect of UV light.
Conclusion and application: The enzyme partially dephosphorylated linearized pUC18 plasmid despite being an alkaline phosphatase and showed potential application as a substitute for alkaline phosphatase from other sources.
Keywords: Vigna unguiculata (Black-eyed Bean), Cajanus indicus (Red Gram), Arachis hypogaea (Ground Nut), alkaline phosphatase, Bradford assay, p-nitrophenylphosphate.
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