Purification and characterization of PAP phytase from Indian wheat variety DBW 17


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Authors

  • Anukriti Verma
  • Sewa Ram
  • Sunita Dalal

https://doi.org/10.25174/61eevc50

Keywords:

Column chromatography, phytase, phytate, purification, wheat whole meal

Abstract

Phytase (myo-inositol hexakisphosphate phosphohydrolase E.C.3.1.3.8) was isolated and purified from Indian wheat variety DBW 17 using 5-step purification procedure as (NH4)2SO4 fractionation, cold methanol fractionation, DEAETrisacryl M chromatography, Ultrogel AcA34 filtration chromatography and SP-Trisacryl M chromatography. Two forms of the enzyme as PHY1 (64 kDa) and PHY2 (62 kDa) were identified. There was 79.0 fold purification of PHY1 and 114.8 fold of PHY2. The optimum pH for PHY1 and PHY2 was 6.0 and 5.5 respectively while optimum temperature 55°C for both the isozymes. The Km value for the hydrolysis of phytate was 123 µM for both the isozymes indicating high specificity for phytate. Wheat phytase can be used in hydrolysis of phytate in food industries for releasing micronutrients and hence increasing their bioavailability

Author Biographies

  • Anukriti Verma

    Directorate of Wheat Research, Agrasain Marg, Post box- 158, Karnal 132 001
  • Sewa Ram

    Directorate of Wheat Research, Agrasain Marg, Post box- 158, Karnal 132 001
  • Sunita Dalal
    Department of Biotechnology, Kurukshetra University, Kurukshetra

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Submitted

2013-11-29

Published

2011-12-30

Issue

Vol. 3 No. 2 (2011)

Section

Research Article

License

Copyright (c) 2014 Journal of Wheat Research

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This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.

The information provided by authors is the sole responsibility by the respective author. The copyright is of the society and no part can be reproduced without prior permission.

How to Cite

Verma, A., Ram, S., & Dalal, S. (2011). Purification and characterization of PAP phytase from Indian wheat variety DBW 17. Journal of Cereal Research, 3(2). https://doi.org/10.25174/61eevc50