Partial Purification of Mulberry (Morus rotunbiloba) Peroxidase Using Aqueous Two-Phase Extraction Coupling with Ion-exchange and Gel-filtration Chromatography

Authors

  • Supannapa Luanghiran Department of General Science, Faculty of Science, Kasetsart University, Bangkok 10900, Thailand.
  • Poontariga Harinnasut Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok 10900, Thailand.
  • Amara Thongpan Department of General Science, Faculty of Science, Kasetsart University, Bangkok 10900, Thailand.
  • Amonrat Proomboon Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok 10900, Thailand.
  • Sunanta Ratanapo Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok 10900, Thailand.

Keywords:

aqueous two-phase, mulberry leaf peroxidase, purification

Abstract

The third to fifth leaves of mulberry plant were selected for peroxidase extraction due to its high specific activity comparing to other sets of leaf. The initial enzyme isolation included homogenization and extraction of phenolic compound using aqueous two-phase extraction system consisting of 20/8.9 %(w/v) PEG/(NH)2SO4. This system gave a satisfactorily less partition coefficient (0.018) and also less peroxidase volume (34% of total volume). The purification was performed on a DEAE-Cellulose column and double chromatography of Sephadex G-75. The overall result of mulberry peroxidase purification gave 157.4-folds with 43.4% recovery. The native molecular weight of mulberry calculated from the relative fraction of the standard curve from gel filtration was found to be 29 kDa as which agreed with one band of the molecular weight identified by SDS-PAGE.

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Published

2004-08-30

How to Cite

Supannapa Luanghiran, Poontariga Harinnasut, Amara Thongpan, Amonrat Proomboon, and Sunanta Ratanapo. 2004. “Partial Purification of Mulberry (Morus Rotunbiloba) Peroxidase Using Aqueous Two-Phase Extraction Coupling With Ion-Exchange and Gel-Filtration Chromatography”. Agriculture and Natural Resources 38 (4). Bangkok, Thailand:501-9. https://li01.tci-thaijo.org/index.php/anres/article/view/243199.

Issue

Vol. 38 No. 4 (2004): July-August

Section

Research Article

License

online 2452-316X print 2468-1458/Copyright © 2022. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/),
production and hosting by Kasetsart University of Research and Development Institute on behalf of Kasetsart University.