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1071037 
Journal Article 
Extraction and purification of Ipomoea peroxidase employing three-phase partitioning 
Narayan, AV; Madhusudhan, MC; Raghavarao, KS 
2008 
Yes 
Applied Biochemistry and Biotechnology
ISSN: 0273-2289
EISSN: 1559-0291 
Humana Press Inc., 999 Riverview Dr., Ste. 208 Totowa NJ 07512 USA 
Totowa 
151 
2-3 
263-272 
English 
18369532 
WOS:000260538600016 
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 degrees 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. 
Ammonium Sulfate; Chemical Fractionation; Electrophoresis, Polyacrylamide Gel; Ipomoea; Isoelectric Point; Peroxidase; Plant Leaves; Temperature; tert-Butyl Alcohol; Ipomoea palmata; purification; extraction; chemical constituents of plants; enzyme activity; polyacrylamide gel electrophoresis; butanol; article 
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