Aqueous two-phase system purification for superoxide dismutase induced by menadione from Phanerochaete chrysosporium

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Kavakcioglu B., Tongul B., Tarhan L.

ARTIFICIAL CELLS NANOMEDICINE AND BIOTECHNOLOGY, vol.45, no.2, pp.380-388, 2017 (SCI-Expanded) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 45 Issue: 2
  • Publication Date: 2017
  • Doi Number: 10.3109/21691401.2016.1160404
  • Journal Name: ARTIFICIAL CELLS NANOMEDICINE AND BIOTECHNOLOGY
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.380-388
  • Keywords: Aqueous two-phase systems, partitioning, polymer/salt system, purification, superoxide dismutase, POLY(ETHYLENE GLYCOL), ESCHERICHIA-COLI, MOLECULAR-WEIGHT, OXIDATIVE STRESS, EXTRACTION, RECOVERY, ENZYMES, CHLOROPLASTS, TEMPERATURE, SEPARATION
  • Dokuz Eylül University Affiliated: Yes

Abstract

In the present work, the partitioning behavior of menadione-induced superoxide dismutase (SOD; EC 1.15.1.1), an antioxidant enzyme that has various applications in the medical and cosmetic industries, from the white rot fungus Phanerochaete chrysosporium has been characterized on different types of aqueous two-phase systems (ATPSs) (poly(ethylene glycol)/polypropylene glycol (PEG/PPG)-dextran, PEG-salt and PPG-salt). PEG-salt combinations were found most optimal systems for the purification of SOD. The best partition conditions were found using the PEG-3350 24% and K2HPO4 5% (w/w) with pH 7.0 at 25 degrees C. The partition coefficient of total SOD activity and total protein concentration observed in this system were 0.17 and 6.65, respectively, with the recovery percentage as 78.90% in the bottom phase and 13.17% in the top phase. The highest purification fold for SOD from P. chrysosporium was found as 6.04 in the bottom phase of PEG 3350% 24 - K2HPO4% 5 (w/w) system with pH 7.0. SOD purified from P. chrysosporium was determined to be a homodimer in its native state with a molecular weight of 60 +/- 64 kDa. Consequently, simple and only one step PEG-salt ATPS system was developed for SOD purification from P. chrysosporium.