title: Treatment of phenolic wastewater by horseradish peroxidase immobilized by bioaffinity layering.
abstract: Horseradish peroxidase was immobilized by bioaffinity layering and used for the treatment of wastewater containing p-chlorophenol. For this purpose, lectin Concanavalin A was bound to Sephadex beads. The glycoenzyme peroxidase was layered upon this Con A layer. Subsequently, alternate layers of the enzyme and Con A were applied. The most efficient design consisted of three layers of Con A and peroxidase each. This immobilized enzyme preparation retained 80% of the activity of the free peroxidase used for immobilization. PEG at the concentration of 0.1 mg ml(-1) was found to prevent enzyme inactivation by the products, although it increased the process time. Thus 60 U ml(-1) of enzyme completely converted the p-chlorophenol (into products) in 4 min in the absence of PEG. On the other hand, only 0.05 U ml(-1) of enzyme was required for this purpose in the presence of PEG but the process required 60 min. Peroxidase converts phenol molecules into free radicals. These free radicals then polymerize and get precipitated. As a further means of minimizing exposure of the enzyme to free radicals and enhancing the reusability, it was decided to remove the enzyme from reaction medium after 10 min. With this strategy, the bioaffinity layered peroxidase preparation could be reused five times without any loss of activity.
authors.affiliation: Department of Chemistry, Indian Institute of Technology, Delhi, Hauz Khas, New Delhi 110 016, India.
authors.forename: Sohel
authors.initials: S
authors.lastname: Dalal
authors.forename: Munishwar Nath
authors.initials: MN
authors.lastname: Gupta
mesh_terms: D002733:Chlorophenols / Q000032:analysis / Q000737:chemistry; D003208:Concanavalin A / Q000032:analysis; D004800:Enzymes, Immobilized / Q000378:metabolism*; D006735:Horseradish Peroxidase / Q000378:metabolism*; D010636:Phenols / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000494:pharmacology; D014865:Waste Disposal, Fluid / Q000379:methods; D018508:Water Purification / Q000379:methods*
chemical_list: D002733:Chlorophenols; D004800:Enzymes, Immobilized; D010636:Phenols; D003208:Concanavalin A; C029107:4-chlorophenol; D011092:Polyethylene Glycols; D006735:Horseradish Peroxidase