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Extraction and Purification of Ipomoea Peroxidase Employing Three-phase Partitioning

Narayan, A. V. and Madhusudhan, M. C. and Raghavarao, K. S. M. S. (2008) Extraction and Purification of Ipomoea Peroxidase Employing Three-phase Partitioning. Applied Biochemistry and Biotechnology, 151. pp. 263-272.

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Abstract

Three-phase partitioning (TPP) is a novel separation process used for the extraction and purification of biomolecules. The biomolecules are recovered in a purified form at the interface (precipitate), while the contaminants partition in t-butanol and aqueous phases. Peroxidase from the leaves of Ipomoea palmata was purified by using TPP. The ratio of the crude extract to t-butanol of 1:1 and 30% ammonium sulfate at 37 °C resulted in about 160% activity recovery and twofold purification in the aqueous phase of the first cycle of TPP. On subjecting the aqueous phase to the second cycle of TPP, a purification of 18-fold was achieved with about 81% activity recovery. The sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis showed substantial purification, and the molecular weight of peroxidase was found to be 20.1 KDa. The present study shows a higher degree of purification and activity yield as a primary purification process in comparison with existing literature values, thus demonstrating TPP as an attractive downstream process for the purification of peroxidase.

Item Type: Article
Uncontrolled Keywords: Three-phase partitioning . Ipomoea palmata . Peroxidase . Cross-partitioning . Aqueous two-phase extraction . Downstream process . t-Butanol . Ammonium sulfate
Subjects: 500 Natural Sciences and Mathematics > 10 Plants
600 Technology > 08 Food technology > 05 Processing and Engineering
Divisions: Food Engineering
Depositing User: Food Sci. & Technol. Information Services
Date Deposited: 07 Jun 2011 06:05
Last Modified: 28 Dec 2011 10:23
URI: http://ir.cftri.com/id/eprint/10073

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