Rekha Kansal Author

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Isolation and Characterization of Protease Inhibitor Protein from Pigeon pea (Cajanus cajan L.)

Author(s):

Mukesh Kumar, Rekha Kansal, Vinay Kumar, P S Srivastava and K R Koundal

Protease Inhibitor (PI) protein was isolated from four varieties of Pigeonpea viz. Pusa-855, Pusa- 33, Pusa-987 ND Pusa-84 by shaking defatted flour of mature seeds with 0.1 M sodium phosphate buffer (pH 7.6). The crude extracts were shacked for four hours at RT in a shaker. The homogenates were heat denatured and centrifuged. The supernatants subjected to 0-20%, 20- 40%, 40-60%, 60-80% and 80-100% ammonium sulphate fractions and the precipitates obtained were dialyzed extensively with same buffer. The dialyzed sample of Pusa-33, which had higher PI activity in 40-80% fraction, was used for further purification. The dialyzed protein sample loaded on pretreated DEAE-cellulose (anion exchanger) column to purify the protein followed by gel filtration. The fract... view more»

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