Application of Antioxidant Enzyme Activity and Biochemical Characterization in Static and Suspension Cultures of Withania somnifera L. towards Food Technology

Abstract

Withania somnifera (L.) Dunal is an erect evergreen shrub commonly known as Ashwagandha. It is widely used in Ayurvedic, and in the traditional pharmacopeia system of India. It is one of the major ingredients in many formulations prescribed for a variety of musculo-skeletal conditions including arthritis and rheumatism. In the present study, variations in the quality and quantity of proteins and antioxidant enzymes were evaluated biochemically and enzymatically from the static and suspension cultures of Withania somnifera L. The nodal segments provided the maximum callusing of 90.25±0.06% with (1mg/l) BAP and (2mg/l) Kn of 2, 4-D. The static and suspension cultures were taken for the analysis of total soluble protein, and screened for antioxidant enzyme activity [catalase (CAT), superoxide dismutase (SOD) and guaiacol peroxidase (GPX)]. The protein content (1.2016 μg/μl) was found to be higher in static culture samples (0.870 μg/μl) than the protein obtained from the suspension culture. The antioxidant enzyme activity (CAT, SOD and GPX) was higher in the static culture samples (301.01± 0.42, 198.92 ± 0.29, 103.75 ± 0.11 nkat/mg of protein) than in the suspension culture. Specific activity staining of isoenzyme pattern exhibited three isoforms (CAT 1, CAT 2 and CAT 3) in the static culture samples but CAT 1 was absent in the samples extracted from the suspension cultures. In case of SOD, four bands (SOD 1, SOD 2, SOD 3 and SOD 4) were found in both samples whereas the intensity of GPX activity was found to be more in the static culture; however, both samples exhibited three isoforms (GPX 1, GPX 2 and GPX 3). Supplementation of the required nutrients along with phytohormones under in vitro conditions might be an enhancing factor to yield antioxidant enzymes in the static culture samples.