1 2 4 3 1 6 8 2 8 6 5 2 1 2 g
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Journal of Chromatography B
Volumes 1114–1115,2019, Pages 86-92

Purification of peroxidase enzyme from radish species in fast and high yield with affinity chromatography technique

Aykut Oztekina, Zuleyha Almazb, Serpil Gernic, Deniz Erelc, Seyma Mehtap Kocakc, Mehmet Emin Sengülc, Hasan Ozdemirc

Department of Medical Services and Techniques, Agri Ibrahim Cecen University Vocational School of Health Services, 04100 Agri, Turkey.

Abstract

In this study, an effective single step affinity method is presented for purifying plant peroxidase (POD) enzymes from radish species. This method make possible to purify the enzymes in high yield and purity. Briefly, 10 different 4-amino benzohydrazide derivatives were synthesized and identified as new competitive POD inhibitors. Then, these derivatives were coupled to Sepharose 4B-L-Tyrosine support matrix by diazotization to form the affinity gels. Purification factors were recorded as 54.8% yield - 665-fold, 33.8% yield - 613-fold, 22.7% yield - 595-fold, 34.4% yield - 781-fold, 40.9% yield - 282-fold for turnip (T-POD), black radish (BR-POD), daikon (D-POD), sweet radish (SR-POD) and kohlrabi radish, (KR-POD), respectively. It has also been shown that the affinity gels, which prepared using the 4-amino 3-bromo benzohydrazide and 4-amino 2-nitro benzohydrazide molecules, capable to purify all radish species POD enzymes in high purity and yield.

Keywords: Peroxidase, Purification, Radish, Affinity chromatography.

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