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All Journal 3BIO: Journal of Biological Science, Technology and Management Current Research on Biosciences and Biotechnology
Niloperbowo, Wardono
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Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Immunology & microbiology Materials Science & Nanotechnology Medicine & Pharmacology Neuroscience

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Journal : 3BIO: Journal of Biological Science, Technology and Management

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The Influence of Polyethylene Glycol Precipitation Methods on Yield and Purity of White Radish Peroxidase Faizah, Nur Al; Giri-Rachman, Ernawati Arifin; Niloperbowo, Wardono
3BIO: Journal of Biological Science, Technology and Management Vol. 6 No. 2 (2024)
Publisher : School of Life Sciences and Technology, Institut Teknologi Bandung

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.5614/3bio.2024.6.2.5

Abstract

Proteins are widely used in various industries as highly valued biotechnology products. One example is horseradish peroxidase isolated from horseradish (Armoracia rusticana) that used as enzyme label in immunochemistry. However, the cultivation of horseradish is limited to subtropical countries, making the dependency on horseradish peroxidase unsustainable for tropical countries. Numerous studies have explored alternative peroxidases, and white radish peroxidase isolated from Raphanus sativus L. has emerged as a promising candidate. In this study, white radish peroxidase is isolated using the polyethylene glycol (PEG) precipitation method which is widely used as a simple and cost-effective method. This study aims to evaluate the effectiveness of the one-step and two-step PEG precipitation method. The one-step PEG precipitation method used in this study was done by mixing the white radish juice with PEG 6000 30% (w/v), while the two-step method was done by mixing it with PEG 400 20% (w/v) and PEG 6000 30% (w/v) consecutively. This study compares the yield and recovery levels of total protein and white radish peroxidase, as well as the enzymatic specific activity of white radish peroxidase isolated both by the one-step PEG precipitation and the two-step PEG precipitation. The results indicate that both extraction methods yield the same level of white radish peroxidase. However, they differ in terms of purity. The two-step extraction method results in white radish peroxidase with higher purity, as evidenced by its specific activity towards the chromogen ABTS in the presence of H2O2.
Co-Authors A'yun, Ramadhani Qurrota ERNAWATI ARIFIN GIRI-RACHMAN Faizah, Nur Al Giri-Rachman , Ernawati Arifin Hakim, Meutia Diva Marselina Tan Nugrahapraja, Husna Sihombing, Rolina Anna Erica