THE POTENTIAL OF RECOMBINANT MANGANESE SUPEROXIDE DISMUTASE OF STAPHYLOCOCCUS EQUORUM AS AN AGENT TO ACCELERATE WOUND HEALING

• Main Author: Annisa Masruroh, Mutiara
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/71094
• Institution: Institut Teknologi Bandung

Reactive oxygen species (ROS) are products of cell metabolism which, in excessive levels can cause oxidative stress and have implications for impairment and delay in wound healing. Recombinant manganese superoxide dismutase enzyme from Staphylococcus equorum (rMnSODSeq) plays a role in maintaining homeostasis of ROS levels by reducing the production of superoxide radical anions (O2•?). Pharmaceutical Biotechnology Laboratory School of Pharmacy ITB has succeeded in designing the formation of rMnSODSeq K38R-A121E, K38R-A121Y, and L169W mutants to increase protein stability against UVC exposure and high temperatures. This study aims to evaluate the potential of rMnSODSeq K38R-A121E, K38R-A121Y, and L169W as active substances in accelerating wound healing through wound healing scratch assays on 3T3 fibroblast cells. Pure protein was first tested for its stability by zymography and colorimetry methods. A stability test against UVC for 45 minutes showed that wild-type rMnSODSeq, K38R-A121E, K38R-A121EY, and L169W were still able to maintain their activity above 88%. Whereas in the stability test at 60ºC for 15 minutes, the K38R-A121E mutant had the highest residual activity of 97.2% compared to wild-type rMnSODSeq, K38R-A121Y, and L169W (75.1%; 15.3%; 48.1% respectively). Cell viability test results showed that wild-type rMnSODSeq, K38R-A121E, and K38R-A121Y, before and after being heated at 50ºC for 1 hour were not toxic and still able to protect 3T3 cells from exposure to UVC for 3 hours. The highest percentage of wound closure (53%) in 24 hours given by K38R-A121E. This finding shows that the K38R-A121E mutant has the best stability and ability as a therapeutic candidate to accelerate wound healing.