Simultaneous purification and immobilization of bitter gourd (Momordica charantia) peroxidases on bioaffinity support

• Published in: Journal of chemical technology and biotechnology (1986) Vol. 80; no. 2; pp. 198 - 205
• Main Authors: Akhtar, S, Khan, A.A, Husain, Q
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 01-02-2005; Wiley
• Subjects: adsorbents; adsorption; bioaffinity support; bioaffinity supports; Biological and medical sciences; Biotechnology; bitter gourd; concanavalin A; Fundamental and applied biological sciences. Psychology; immobilization; Immobilization of enzymes and other molecules; Immobilization techniques; immobilized enzymes; Methods. Procedures. Technologies; Momordica charantia; peroxidase; peroxidases; phenol removal; remediation; stabilization; wastewater; Purification; Stability; Enzyme; Immobilization; Biological purification; Peroxidases; Affinity; Peroxidase; Oxidoreductases; Adsorbent; Waste water purification; Application; Immobilized enzyme
• ISSN: 0268-2575; 1097-4660
• DOI: 10.1002/jctb.1179

This paper demonstrates the construction of an inexpensive bioaffinity adsorbent by simply incubating Sephadex G 50 matrix with jack bean meal extract at room temperature. Sephadex G 50 adsorbed 17 mg Con A (concanavalin A) per g of the matrix. Con A‐adsorbed Sephadex was employed for the immobilization of glycoenzymes directly from ammonium sulfate‐fractionated proteins of bitter gourd. The obtained bioaffinity support was very efficient for high yield immobilization of peroxidases from bitter gourd and it bound nearly 425 enzyme units per g of the matrix. Bitter gourd peroxidase immobilized on lectin–Sephadex support showed a very high effectiveness factor, ‘η,’ of 1.25. Immobilized BGP preparation was quite stable against the denaturation induced by pH, heat, urea, Triton X 100, Tween 20, SDS, Surf Excel and water‐miscible organic solvents: dimethyl sulfoxide and dimethyl formamide. Low concentration of detergents like SDS, Tween 20, and Triton X 100 enhanced the activity of soluble and immobilized bitter gourd peroxidase. Peroxidase bound to the bioaffinity support exhibited very high resistance to proteolysis caused by the trypsin treatment. Con A–Sephadex‐bound bitter gourd peroxidase retained 85% of its initial activity after treatment with 2.5 mg trypsin per cm3 of incubation mixture for 1 h at 37 °C while the soluble enzyme lost nearly 40% of the initial activity under similar incubation conditions. Immobilized bitter gourd peroxidase preparation appeared to be more rigid to proteolysis mediated by trypsin compared with soluble bitter gourd peroxidase. Copyright © 2004 Society of Chemical Industry