Extraction and purification of Ipomoea peroxidase employing three-phase partitioning

Narayan, AV; Madhusudhan, MC; Raghavarao, KS

HERO ID

1071037

Reference Type

Journal Article

Year

2008

Language

English

PMID

18369532

HERO ID 1071037
In Press No
Year 2008
Title Extraction and purification of Ipomoea peroxidase employing three-phase partitioning
Authors Narayan, AV; Madhusudhan, MC; Raghavarao, KS
Journal Applied Biochemistry and Biotechnology
Volume 151
Issue 2-3
Page Numbers 263-272
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 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.
Doi 10.1007/s12010-008-8185-4
Pmid 18369532
Wosid WOS:000260538600016
Url https://www.proquest.com/scholarly-journals/extraction-purification-ipomoea-peroxidase/docview/48174081/se-2?accountid=171501
Is Certified Translation No
Dupe Override No
Is Public Yes
Language Text English
Keyword 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
Is Qa No