Peroxidase Case Study

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6.1 Introduction
The peroxidase isolated from horseradish, HRP, is the most available and commonly used peroxidase. One factor that has limited its widespread and large-scale use is its high cost of production. A cost effective purification technology and exploringalternative sources with high peroxidase activity can help to bring down the cost of enzyme production. Peroxidase from roots of Raphanus sativus can serve as a cost effective alternative for HRP.
Gil-Rodríguez et al. (2008) purified the peroxidase from Japanese radish by several purification steps. The juice extracted from roots of radish was ultra-centrifuged using 10 kDa membrane. The retentate was then loaded on Macro-Prep High-S medium at pH 6.1 and eluted with a linear gradient of 0–1 M NaCl. The eluate was further loaded on Macro-Prep High-Q medium using similar protocol and the eluate so obtained was allowed to equilibrate overnight and later loaded onto third column Macro-Prep CM at pH 4 and eluted with linear gradient of 0-1 M NaCl. The active fractions were concentrated in an Amicon ultrafiltration cell using a 10 kDa cutoff membrane.
Ashraf and Hussain (2010) had attempted crude fractionation of radish peroxidase using ammonium sulphate. This technique has its own limitations and hence not used in commercial manufacturing. Aruna and Lali (2001) attempted to isolate peroxidase from the roots of Raphanus sativus using CM cellulose in the form of soluble ion-exchange macroligand to precipitate most

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