IN VITRO EVALUATION OF PARTIALLY PURIFIED ANTIOXIDANT ENZYMES FROM LICHEN LEPTOGIUM PAPILLOSUM

ABSTRACTObjective: In this study, the enzyme activity of the partially purified six different antioxidant enzymes (AEs) of Leptogium papillosum such assuperoxide dismutase (SOD), catalase (CAT), peroxidase (POD), polyphenol oxidase (PPO), glutathione transferase (GST), and glutathione peroxidase(GPx) was carried out.Methods: Ammonium sulfate precipitation, dialysis, and DEAE-cellulose column chromatography were the three different methods performed topurify the six different AEs from L. papillosum. The antioxidant enzyme activity of the purified extracts was determined in vitro by the followingstandard procedure of SOD, CAT, POD, PPO, GST, and GPx.Results and Conclusion: The results revealed that there was a significant increase (p<0.001) in the specific activity of purified fractions of all theenzymes with a corresponding increase in the purification fold. The comparable activity of PPO and GST were determined using cluster analysis usingshort linkage distance. Principal component analysis indicated that SOD contributed primarily to the total variation in the AEs among allthe other fractions with the specific activity of 21.70 U/mg by attaining the purification fold of 5.91. Thus, our findings suggested that the purifiedAEs of L. papillosum possess potent antioxidant defense machinery by scavenging free radical population. Moreover, SOD was played a major role ofcapturing free radical by having highest enzyme activity followed by GST and CAT.DEAE-celluloseKeywords: Antioxidant enzymes, Cluster analysis, DEAE-cellulose column chromatography, Lichen, Leptogium papillosum, Principal componentanalysis

IN VITRO ASSESSSMENT OF ANTIOXIDANT AND ANTIMICROBIAL ACTIVITIES OF DIFFERENT SOLVENT EXTRACTS FROM LICHEN RAMALINA NERVULOSA

Objective: The purpose of the existent study is to assess the in vitro antioxidant and antimicrobial activity of different solvent extracts of Ramalina nervulosa. Methods: Folin ciocalteu's and Aluminium chloride method were used for total phenolic and flavonoid content estimation respectively. The antioxidant activity was evaluated through free radical scavenging activity by DPPH (2, 2-diphenyl-1-picrylhydrazyl), FRAP (ferric ions reducing antioxidant potential), cupric ion chelation ability. Agar diffusion method was used to assess the antimicrobial capacity. GC-MS analysis was carried out to identify the presence of compounds in the extracts.Results: Ethanol extracts exhibited that the highest zone of inhibition with Bacillus subtilis, Proteus vulgaris, Pseudomonas aeruginosa, Klebsiella pneumoniae and with two fungal strains such as Fusarium oxyparum and Penicillium notatum. The extract also has high inhibition capacity of 64.29% and 62.49% towards DPPH and Cu2+ion respectively. But the aqueous extract showed higher amount of phenolic and flavonoid content of about 211.40 mg GAE/g dw and 50.72 mg QE/g dw respectively and Fe2+ ion reducing capacity of about 144.41 mol ascorbic acid equivalent/g db. Further analysis of GC-MS showed the presence of various bioactive molecules present in the lichen Ramalina nervulosa.Conclusion: The results of FRAP assay were positively correlated with the total phenolic and flavonoid content. Ethanol extracts of R. nervulosa showed promising antibacterial, antifungal and antioxidant activities which can be considered as the potent source of antioxidant product and offer protection from oxidative stress under many pathophysiological conditions.Â