This work presents a novel approach for the recovery and purification of superoxide dismutase (SOD; EC 22.214.171.124) produced in the yeast Kluyveromyces marxianus var. bulgaricus (NBIMCC 1984). Electroextraction was evaluated as an alternative to bead milling for SOD release. Seven anion exchange resins were studied in order to optimize SOD purification using ion exchange chromatography.
Electroextraction produced an enzyme solution with a specific activity 13 times higher than with mechanical extraction, demonstrating the potential of this novel technique for the selective liberation of SOD. Regarding the anion exchange resins that were studied, Amberlite™ IRA-96 and Q Sepharose™ XL were chosen as the best options for purifying SOD using chromatography in negative adsorption mode. The process developed, involving electroextraction, enzyme concentration and negative anion exchange chromatography, rendered a highly active and pure SOD preparation (specific activity: 2056 U mg−1 and 66-fold purification) with an overall recovery yield of 84%.
The results presented herein demonstrate the potential of the process developed for the recovery and purification of SOD from the yeast Kluyveromyces marxianus var. bulgaricus (NBIMCC 1984). The preparation obtained could be used in the food industry as an antioxidant agent. © 2013 Society of Chemical Industry