Purification, characterization and selected inhibition properties of peroxidase from haricot bean (Phaseolus vulgaris L.)

Koktepe T., Altin S., Tohma H., GÜLÇİN İ., KÖKSAL E.

INTERNATIONAL JOURNAL OF FOOD PROPERTIES, cilt.20, ss.1944-1953, 2017 (SCI-Expanded) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 20
  • Basım Tarihi: 2017
  • Doi Numarası: 10.1080/10942912.2017.1360903
  • Dergi Adı: INTERNATIONAL JOURNAL OF FOOD PROPERTIES
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.1944-1953
  • Anahtar Kelimeler: Haricot bean (Phaseolus vulgaris), Peroxidase, Enzyme purification, Enzyme characterization, Enzyme inhibition, HORSERADISH-PEROXIDASE, ANIONIC PEROXIDASE, ISOENZYMES, LEAVES
  • Atatürk Üniversitesi Adresli: Evet

Özet

In this study, a cationic soluble peroxidase (POD) was homogeneously purified and biochemically characterized. The enzyme purification involved the combination of (NH4)(2)SO4 precipitation, CM-Sephadex cation exchange, and gel filtration chromatography. The purity of the enzyme was examined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, and one single prominent band was obtained with a molecular weight of approximately 45 kDa. The thermal and pH stability showed that the enzyme was stable at pH 5.0 and at 40 degrees C, respectively. The activity remained stable at pH 4.0 at least for 10days. K-m and V-max values were calculated from Lineweaver-Burk graph for each substrate. POD exhibited K-m values of 0.0154 and 0.065mM for guaiacol and H2O2, respectively. The enzyme was highly inhibited by sodium azide. This study could enrich the literature regarding the properties of POD from haricot bean that could be applicable to biomedical analysis.