Extraction and purification of a plant peroxidase by aqueous two-phase extraction coupled with gel filtration

Srinivas, N. D. and Rashmi, K. R. and Raghavarao, K. S. M. S. (1999) Extraction and purification of a plant peroxidase by aqueous two-phase extraction coupled with gel filtration. Process Biochemistry (35). pp. 43-48.

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Abstract

A plant peroxidase extracted from leaves of Ipomoea palmetta was partially purified by extraction using a polyethylene glycol (PEG):ammonium sulphate aqueous two-phase system (ATPS), followed by gel filtration on a Sephadex G-100 column. Variation of phase composition and NaCl concentration resulted in the desired reduction in volume of the extract and selective partitioning of the enzyme. PEG:ammonium sulphate:NaCl (24:7.5:2.0%, w:v) system gave a partition coefficient of 0.042, purification factor of 2.18, volume reduction of 57.5% and bottom phase recovery of 91.5%. The enzyme was further concentrated by anti-dialysis and was finally purified about 49-fold by gel filtration on a Sephadex G-100 column, with a recovery of 75.3%. The enzyme was stable at 60°C with an optimum activity at 50°C. The enzyme was stable at pH range 5–11 and had an optimum pH at 6.0. The enzyme showed a Km of 18.15 mM and Vmax of 27.1 mmol:min when guaiacol was used as substrate.

Item Type:	Article
Uncontrolled Keywords:	Plant peroxidase; Aqueous two-phase extraction; Gel filtration
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:	01 Oct 2007 12:32
Last Modified:	11 Oct 2018 06:27
URI:	http://ir.cftri.res.in/id/eprint/1456

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