Partial purification and biochemical characterization of an extremely thermo- and pH-stable esterase with great substrate affinity

Ozbek E., KOLCUOĞLU Y., Konak L., ÇOLAK A., Oz F.

TURKISH JOURNAL OF CHEMISTRY, cilt.38, sa.4, ss.538-546, 2014 (SCI-Expanded) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 38 Sayı: 4
  • Basım Tarihi: 2014
  • Doi Numarası: 10.3906/kim-1308-23
  • Dergi Adı: TURKISH JOURNAL OF CHEMISTRY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, TR DİZİN (ULAKBİM)
  • Sayfa Sayıları: ss.538-546
  • Karadeniz Teknik Üniversitesi Adresli: Evet

Özet

An esterase from a thermophilic bacterium, Geobacillus sp. DF20, was partially purified. Final purification factor was found to be 64.5-fold using Q-Sepharose ion exchange column chromatography. Native polyacrylamide gel electrophoresis indicated the presence of a single active esterase. The substrate specificity of this esterase was high for p-nitrophenyl butyrate (pNPB) substrate. The optimum pH and temperature for the enzyme activity were 7.0 and 50 degrees C, respectively. The pH and heat stability profiles show that this enzyme is more stable under neutral conditions at 50 degrees C. K-m and V-max values for this esterase acting on pNPB were 0.12 mM and 54.6 U/mg protein, respectively. Presence of 10% (v/v) acetonitrile in the reaction medium indicated that purified enzyme was strongly inhibited. It was also detected that some metal ions affected enzyme activity at different rates. As a result, it was observed that esterase from Geobacillus sp. DF20 has extreme temperature and pH stabilities. Therefore, the stability and K-m value of the enzyme make this study interesting when compared with the literature.

An esterase from a thermophilic bacterium, Geobacillus sp. DF20, was partially purified. Final purification factor was found to be 64.5-fold using Q-Sepharose ion exchange column chromatography. Native polyacrylamide gel electrophoresis indicated the presence of a single active esterase. The substrate specificity of this esterase was high for p-nitrophenyl butyrate (pNPB) substrate. The optimum pH and temperature for the enzyme activity were 7.0 and 50 degrees C, respectively. The pH and heat stability profiles show that this enzyme is more stable under neutral conditions at 50 degrees C. K-m and V-max values for this esterase acting on pNPB were 0.12 mM and 54.6 U/mg protein, respectively. Presence of 10% (v/v) acetonitrile in the reaction medium indicated that purified enzyme was strongly inhibited. It was also detected that some metal ions affected enzyme activity at different rates. As a result, it was observed that esterase from Geobacillus sp. DF20 has extreme temperature and pH stabilities. Therefore, the stability and K-m value of the enzyme make this study interesting when compared with the literatüre.