Abstract
This study was aimed on purification, immobilization and application of peroxidase enzyme from whole plant of soy bean for the decolorization of textile dyes and degradation of some phenolic compounds. The purification of peroxidase was in two steps include: concentration by sucrose and gel filtration by using sephadex G-150. The purification fold was 4.46 and 2.09 with an enzyme yield of 22.34% and 25.55% for peak 1 and peak 2 respectively. The purified peroxidase enzyme was immobilized by two methods include: covalent linkage immobilization by gluteraldehyde activated chitosan and entrapment by sodium alginate and agarose gel, the immobilization ratio reached to 58.07%, 50% and 11.75% respectively. Crude, purified and immobilized peroxidase were studied with textile dyes (red, blue, yellow and black) after 24 hr, the maximum removal efficiency was with immobilized peroxidase reached to 13.25, 35.12, 14.37 and 26.92 % respectively. Immobilized peroxidase on chitosan was able to degrade some phenolic compounds (tannic acid, naphthalene and gallic acid) during 2 hr, and the degradation efficiency was reached to 95.85%, 79.75% and 33.88% respectively.
Highlights
The soybean (Glycine max), or soya bean, is a species of legume native to east Asia, widely grown for its edible beans
Peroxidase of whole plant purified by gel filteration chromatography using sephadex G-150 column immobilized using entrapment method(sodium alginate, agarose gel) and covalent linkage by chitosan MATERIALS AND METHODS Purification of peroxidase enzyme Peroxidase was purified from soy bean plant using dialysis method for enzyme concentration, followed by gel filtration
The squares of agarose-bounded enzyme were stored in 0.2 M of sodium phosphate buffer pH 7 at 4o C and the Immobilized enzyme activity was determined Immobilization of peroxidase by covalent method by chitosan One gram of chitosan was added to 10 ml of 2% gluteraldehyde solution, stirred gently and mixed for 2 hr, at 4 o C followed by an overnight incubation
Summary
The soybean (Glycine max), or soya bean, is a species of legume native to east Asia, widely grown for its edible beans. Peroxidase of whole plant purified by gel filteration chromatography using sephadex G-150 column immobilized using entrapment method(sodium alginate, agarose gel) and covalent linkage by chitosan MATERIALS AND METHODS Purification of peroxidase enzyme Peroxidase was purified from soy bean plant using dialysis method for enzyme concentration, followed by gel filtration. The squares of agarose-bounded enzyme were stored in 0.2 M of sodium phosphate buffer pH 7 at 4o C and the Immobilized enzyme activity was determined Immobilization of peroxidase by covalent method by chitosan One gram of chitosan was added to 10 ml of 2% gluteraldehyde solution, stirred gently and mixed for 2 hr, at 4 o C followed by an overnight incubation. The resulted chitosan - gluteraldehyde – peroxidase conjugates were separated and washed and Immobilized enzyme activity was determined [8].
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