Purification and characterization of the carbonic anhydrase enzyme from Black Sea trout (Salmo trutta Labrax Coruhensis) kidney and inhibition effects of some metal ions on enzyme activity

KUCUK M., GÜLÇİN İ.

ENVIRONMENTAL TOXICOLOGY AND PHARMACOLOGY, cilt.44, ss.134-139, 2016 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 44
  • Basım Tarihi: 2016
  • Doi Numarası: 10.1016/j.etap.2016.04.011
  • Dergi Adı: ENVIRONMENTAL TOXICOLOGY AND PHARMACOLOGY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.134-139
  • Anahtar Kelimeler: Black Sea trout, Salmo trutta Labrax, Enzyme purification, Enzyme characterization, Carbonic anhydrase, Heavy metal, ERYTHROCYTES IN-VITRO, ISOENZYMES I, GENE-EXPRESSION, ISOZYMES I, ANTIOXIDANT, DERIVATIVES, SULFAMIDES, SERIES, LACTOPEROXIDASE, MELATONIN
  • Atatürk Üniversitesi Adresli: Evet

Özet

In this study, the carbonic anhydrase (CA) enzyme was purified from Black Sea trout (Salmo trutta Labrax Coruhensis) kidney with a specific activity of 603.77 EU/mg and a yield of 35.5% using Sepharose-4B-L-tyrosine-sulphanilamide affinity column chromatography. For determining the enzyme purity and subunit molecular mass, sodiumdodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) was performed and single band was observed. The molecular mass of subunit was found approximately 29.71 kDa. The optimum temperature, activation energy (E-a), activation enthalpy (Delta H) and Q(10) values were obtained from Arrhenius plot. K-m and V-max values for p-nitrophenyl acetate of the purified enzyme were calculated from Lineweaver-Burk graphs. In addition, the inhibitory effects of different heavy metal ions (Fe2+, Pb2+, Co2+, Ag+ and Cu2+) on Black Sea trout kidney tissue CA enzyme activities were investigated by using esterase method under in vitro conditions. The heavy metal concentrations inhibiting 50% of enzyme activity (IC50) were obtained. Finally K-i values and inhibition types were calculated from Lineweaver-Burk graphs. (C) 2016 Elsevier B.V. All rights reserved.