Enhancement of potency and stability of human extracellular superoxide dismutase

• Published in: BMB reports Vol. 48; no. 2; pp. 91 - 96
• Main Authors: Kim, S., The Catholic University of Korea, Seoul, Republic of Korea, Kim, H.Y., The Catholic University of Korea, Seoul, Republic of Korea, Kim, J.H., The Catholic University of Korea, Seoul, Republic of Korea, Choi, J.H., The Catholic University of Korea, Seoul, Republic of Korea, Ham, W.K., The Catholic University of Korea, Seoul, Republic of Korea, Jeon, Y.J., The Catholic University of Korea, Seoul, Republic of Korea, Kang, H., Incheon National University, Incheon, Republic of Korea, Kim, T.Y., The Catholic University of Korea, Seoul, Republic of Korea
• Format: Journal Article
• Language: English
• Published: Korea (South) Korean Society for Biochemistry and Molecular Biology 01.02.2015; 생화학분자생물학회
• Subjects: Copper - chemistry; Extracellular superoxide dismutase,Metalloenzyme,Reactive oxygen species,Refolding; HEK293 Cells; Humans; INFLAMACION; INFLAMMATION; Polyethylene Glycols - chemistry; Protein Stability; Reactive Oxygen Species - metabolism; Recombinant Fusion Proteins - biosynthesis; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - isolation & purification; Superoxide Dismutase - chemistry; Superoxide Dismutase - genetics; Superoxide Dismutase - metabolism; Zinc - chemistry; 화학
• ISSN: 1976-6696; 1976-670X
• DOI: 10.5483/BMBRep.2015.48.2.093

Cells express several antioxidant enzymes to scavenge reactive oxygen species (ROS) responsible for oxidative damages and various human diseases. Therefore, antioxidant enzymes are considered biomedicine candidates. Among them, extracellular superoxide dismutase (SOD3) had showed prominent efficacy against asthma and inflammation. Despite its advantages as a biomedicine, the difficulty in obtaining large quantity of active recombinant human SOD3 (rhSOD3) has limited its clinical applications. We found that a significant fraction of overexpressed rhSOD3 was composed of the inactive apo-enzyme and its potency against inflammation depended on the rate of metal incorporation. Also, purified rhSOD3 was unstable and lost its activity very quickly. Here, we suggest an ideal preparative method to express, purify, and store highly active rhSOD3. The enzymatic activity of rhSOD3 was maximized by incorporating metal ions into rhSOD3 after purification. Also, albumin or polyethylene glycol prevented rapid inactivation or degradation of rhSOD3 during preparative procedures and long-term storage.