Immobilization of catalase onto Eupergit C and its characterization


ALPTEKİN Ö., TÜKEL S. S., YILDIRIM D., ALAGÖZ D.

JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC, cilt.64, ss.177-183, 2010 (SCI-Expanded) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 64
  • Basım Tarihi: 2010
  • Doi Numarası: 10.1016/j.molcatb.2009.09.010
  • Dergi Adı: JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.177-183
  • Anahtar Kelimeler: Catalase, Eupergit C, Covalent immobilization, Batch type reactor, Plug flow type reactor, CIBACRON BLUE F3GA, ENZYME IMMOBILIZATION, REVERSIBLE IMMOBILIZATION, STABILIZATION, ADSORPTION, ATTACHMENT, SUPPORTS, CHITOSAN
  • Çukurova Üniversitesi Adresli: Evet

Özet

Bovine liver catalase was covalently immobilized onto Eupergit C. Optimum conditions of immobilization: pH, buffer concentration, temperature, coupling time and initial catalase amount per gram of carrier were determined as 7.5, 1.0 M, 25 degrees C, 24h and 4.0 mg/g, respectively. V-max and K-m were determined as 1.4(+/- 0.2) x 10(5) U/mg protein and 28.6 +/- 3.6 mM, respectively, for free catalase, and as 3.7(+/- 0.4) x 10(3) Wing protein and 95.9 +/- 0.6 mM, respectively, for immobilized catalase. The thermal stability of the immobilized catalase in terms of half-life time (29.1 h) was comparably higher than that of the free catalase (9.0 h) at 40 degrees C. Comparison of storage stabilities showed that the free catalase completely lost its activity at the end of 11 days both at room temperature and 5 degrees C. However, immobilized catalase retained 68% of its initial activity when stored at room temperature and 79% of its initial activity when stored at 5 degrees C at the end 01 28 days. The highest reuse number of immobilized catalase was 22 cycles of batch operation when 40 mg of immobilized catalase loaded into the reactor retaining about 50% of its original activity. In the plug flow type reactor, the longest operation time was found as 82 min at a substrate flow rate of 2.3 mL/min when the remaining activity of 40 mg immobilized catalase was about 50% of its original activity. The resulting immobilized catalase onto Eupergit C has good reusability, thermal stability and long-term storage stability. (C) 2009 Elsevier B.V. All rights reserved.