Single-step purification of peroxidase enzyme from Horseradish (Raphanus sativus L.)

The Horseradish peroxidase (HRP) is promising catalyst in the enzymatic process of hydrogen peroxidase (H2O2) by single-step affinity method for purifying. This research used DEAE-Sepharose resin as ion-exchangers due to the most favorable choices that allow simple method of enzyme purification to o...

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Bibliographic Details
Main Authors: Lestari, Fatma Puji, Al-Baarri, Ahmad Ni’matullah, Widayat, Legowo, Anang Mohamad
Format: Conference Proceeding
Language:English
Subjects:
Dilution
Enzyme activity
Enzymes
Hydrogen peroxide
Ion exchangers
Peroxidase
Purification
Resins
Sodium chloride
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Summary:The Horseradish peroxidase (HRP) is promising catalyst in the enzymatic process of hydrogen peroxidase (H2O2) by single-step affinity method for purifying. This research used DEAE-Sepharose resin as ion-exchangers due to the most favorable choices that allow simple method of enzyme purification to obtain crude enzyme in mass volume. As much as 100 ml of local radish extract was eluted to the column containing resin and the fraction was then collected using serial dilution with 0.4 and 1 mM sodium chloride. Ten fractions were collected into each tube until it was reached 10 mL of total volume. The result shown that the highest absorbance value was obtained at fifth fraction with enzyme activity 52.52 units/mL in the dilution using 0.4 mM while no peak was detected at the dilution using 1 mM sodium chloride. Based on the results, it is concluded that crude HRP could be obtained using ion-exchanger via single-step purification. This may allow the industry to get the HRP in a simple way of purification.
ISSN:0094-243X
1551-7616
DOI:10.1063/1.5140942