Purification and Immobilization of Superoxide Dismutase Obtained from Saccharomyces cerevisiae TBRC657 on Bacterial Cellulose and Its Protective Effect against Oxidative Damage in Fibroblasts

Phitsanu Pinmanee; Kamonwan Sompinit; Angkana Jantimaporn; Mattaka Khongkow; Dietmar Haltrich; Thidarat Nimchua; Prakit Sukyai

doi:10.3390/biom13071156

Purification and Immobilization of Superoxide Dismutase Obtained from Saccharomyces cerevisiae TBRC657 on Bacterial Cellulose and Its Protective Effect against Oxidative Damage in Fibroblasts

Biomolecules. 2023 Jul 20;13(7):1156. doi: 10.3390/biom13071156.

Authors

Phitsanu Pinmanee^{1

2}, Kamonwan Sompinit², Angkana Jantimaporn³, Mattaka Khongkow³, Dietmar Haltrich⁴, Thidarat Nimchua², Prakit Sukyai¹

Affiliations

¹ Biotechnology of Biopolymers and Bioactive Compounds Special Research Unit, Department of Biotechnology, Faculty of Agro-Industry, Kasetsart University, Bangkok 10900, Thailand.
² Enzyme Technology Research Team, National Center of Genetic Engineering and Biotechnology (BIOTEC), Pathum Thani 12120, Thailand.
³ Nanomedicine and Veterinary Research Team, National Center of Nanotechnology (NANOTEC), Pathum Thani 12120, Thailand.
⁴ Department for Food Science and Food Technology, University of Natural Resources and Life Sciences (BOKU), 1190 Vienna, Austria.

Abstract

Superoxide dismutase (SOD) is an essential enzyme that eliminates harmful reactive oxygen species (ROS) generating inside living cells. Due to its efficacities, SOD is widely applied in many applications. In this study, the purification of SOD produced from Saccharomyces cerevisiae TBRC657 was conducted to obtain the purified SOD that exhibited specific activity of 513.74 U/mg with a purification factor of 10.36-fold. The inhibitory test revealed that the purified SOD was classified as Mn-SOD with an estimated molecular weight of 25 kDa on SDS-PAGE. After investigating the biochemical characterization, the purified SOD exhibited optimal activity under conditions of pH 7.0 and 35 °C, which are suitable for various applications. The stability test showed that the purified SOD rapidly decreased in activity under high temperatures. To overcome this, SOD was successfully immobilized on bacterial cellulose (BC), resulting in enhanced stability under those conditions. The immobilized SOD was investigated for its ability to eliminate ROS in fibroblasts. The results indicated that the immobilized SOD released and retained its function to regulate the ROS level inside the cells. Thus, the immobilized SOD on BC could be a promising candidate for application in many industries that require antioxidant functionality under operating conditions.

Keywords: ROS elimination; bacterial cellulose; enzyme immobilization; enzyme purification; superoxide dismutase.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Fibroblasts / metabolism
Oxidative Stress
Reactive Oxygen Species
Saccharomyces cerevisiae* / metabolism
Superoxide Dismutase* / metabolism

Substances

Reactive Oxygen Species
Superoxide Dismutase

Grants and funding

This research received funding by National Center for Genetic Engineering and Biotechnology (BIOTEC) under a research project “Development of Specialty Enzymes for Eco-Industrial Innovative Products” (grant number: P-18-50932), Thailand Graduate Institute of Science and Technology (TGIST) by National Science and Technology Development Agency (NSTDA) (grant number: TG-22-11-60-007D) and Biotechnology of Biopolymers and Bioactive Compounds Special Research Unit, Department of Biotechnology, Faculty of Agro-Industry, Kasetsart University.