Teknologi Indonesia
Vol 32, No 2 (2009)

PURIFICATION AND CHARACTERIZATION OF WHITE RADISH (RAPHANUS SATIVUS L.VAR LONG WHITE) PEROXIDASE FROM CELL CULTURE EXTRACT

Pudjiraharti, - (Unknown)
Karossi, A.T. (Unknown)



Article Info

Publish Date
28 Nov 2012

Abstract

Horseradish peroxidase (HRP) has been widely used as a component of clinical diagnostic reagent for Enzyme Linked I mmunosorbent Assay (ELISA) technique. White radish as one of Brassicaceae family members was found to have high peroxidase activity. In this study, white radish (Raphanus sativus L. var Long White) was used for the production of peroxidase by cell suspension culture technique. Isolation of the enzyme was conducted by ammonium sulfate precipitation followed by purification using DEAE-Cellulose column chromatography eluted with 0.01 M phosphate buffer, pH 7.5 and 0-0.5 M NaCl gradient. A major peak of protein having the highest enzyme activity and purity 25 folds compared to the crude was resulted. SDS-Polyacrilamide gel electrophoresis showed one main band with molecular weight of 47.000 Da. This white radish peroxidase enzyme is very efficient by demonstrating maximum activity at temperature 55oC and pH 7.5 as well as a Km 76.6 ?g/mL and V max 275 ?g/mL/minute toward hydrogen peroxide substrate and pyrogallol hydrogen donor.

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Journal Info

Abbrev

JTI

Publisher

Subject

Computer Science & IT

Description

JTI is a journal in the Departement of Engineering Sciences - Indonesian Institute of Sciences (LIPI). JTI has policy to publish a new and original research paper or a review paper in The scope of Technology. JTI publishes two issues per year. The journal has been registered with printed-ISSN ...