Purification of tea leaf (Camellia Sinensis) polyphenol oxidase by using affinity chromatography and investigation of its kinetic properties"

Yılmaz C., Aksoy M., Küfrevioğlu Ö. İ.

JOURNAL OF FOOD MEASUREMENT AND CHARACTERIZATION, cilt.15, ss.1-6, 2019 (SCI-Expanded)

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 15
  • Basım Tarihi: 2019
  • Dergi Adı: JOURNAL OF FOOD MEASUREMENT AND CHARACTERIZATION
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.1-6
  • Atatürk Üniversitesi Adresli: Evet

Özet

Polyphenol oxidase (PPO) was purified from tea leaves (Camellia sinensis) via affinity chromatography for the first time

and the purified enzyme was characterized. The purity of enzyme and molecular weight was determined by SDS-PAGE

and non-denaturing PAGE (native PAGE). Single bands were observed with both electrophoretically methods. The PAGE

results indicated that the molecular weight of PPO from tea leaf was approximately 50 kDa. The pH, temperature, and

kinetic parameters were studied. Km

values were 3.782 and 3.881 mM for catechol and 4-methylcatechol respectively. Vmax

values for catechol and 4-methylcatechol were also determined as 1.676 and 1.912 μmol/L min respectively. Additionally,

the inhibition effects of sodium metabisulfite, sodium sulfite, ascorbic acid, glutathione, dithioerythritol were investigated

on the enzyme activity and IC50,

Ki

values were calculated for these substances.