ARTI KUMARI, L. K. CHUGH*, VINOD KUMAR, SUSHIL NAGAR, NEERAJ KHAROR AND SHEENU
Department of Biochemistry, College of Basic Sciences & Humanities,
CCS Haryana Agricuultural University, Hisar-125 004 (Haryana), India
*(e-mail : firstname.lastname@example.org)
(Received : 14 June 2020; Accepted : 29 June 2020)
Pearl millet [Pennisetum glaucum (L.) R. Br.] is an important food and fodder crop, grown in arid and semi-arid tropical regions. It has great nutritional significance, however, the high amount of phytic acid is considered as anti-nutritional compound towards its use in food/feed for monogastric animals. Hydrolysis of phytate using phytase enzyme is a promising approach for reducing phytate content. In this study, a pearl millet genotype HC 20 with phytase activity of ~63.3 µg Pi/h/g was used for phytase extraction, partial purification and study of properties of enzyme. Phytase was partially purified by ammonium sulphate precipitation (30-80% saturation) upto 3.8 fold recovering 54.5 % of the activity. The purified preparation showed maximum activity at pH 5.0. Activities of the crude and ammonium sulphate fractions were 3589 µg Pi/h and 1958 µg Pi/h, respectively. The Km of purified enzyme was 1.51 mM phytate. The purified preparation exhibited optimum activity at 50°C. The enzyme was stable up to 60°C for 30 min. The study revealed the properties of enzyme which might be helpful for its efficient application during food/feed processing for enhanced quality.
Key words:Phytase, phytate, partial purification, characterization, enzyme kinetics