Evaluation of Antioxidant Enzyme Activity and Biochemical Characterization from Static and Suspension Culture of Withania somnifera L

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 musculoskeletal conditions including arthritis and rheumatism. In the present study the variation in quality and quantity of protein and antioxidant enzymes were evaluated biochemically and enzymatically from the static and suspension cultures. The nodal segments had provided maximum callusing of 90.25±0.06 % with (1mg/l) of BAP and Kn with (2mg/l) 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 static culture samples (301.01± 0.42, 198.92 ± 0.29, 103.75 ± 0.11 nkat/ mg of protein) than that of suspension culture. Specific activity staining of isozyme pattern exhibited three isoforms (CAT 1, CAT 2 and CAT 3) in static culture samples but CAT 1 was absent in the sample extracted from suspension cultures. In case of SOD, four bands (SOD 1, SOD 2, SOD 3 and SOD 4) were found in both the samples whereas intensity of GPX activity was found to be more in static culture but both the samples exhibited three isoforms such as (GPX 1, GPX 2 and GPX 3). The supplementation of required nutrients along with the phytohormones under in vitro condition might be an enhancing factor to yield antioxidant enzymes in the static culture samples.