REVERSE AGING IN SACCHAROMYCES CEREVISIAE THROUGH REDUCED TOR1 EXPRESSION AND REPLICATIVE LIFE SPAN MODELLING WITH CELL VIABILITY ASSESSMENT USING PHYTOCHEMICAL EXTRACTS ALPINIA GALANGAL, COMMIPHORA MUKUL, ACORUS CALAMUS AND MORSHELLA ESCULENTA

To study reverse aging in Saccharomyces cerevisiae (S. cerevisiae) by gene expression analysis of target of rapamycin (TOR1) induced autophagy and replicative life span modelling (RLS) using Alpinia galangal (A. galangal), Commiphora mukul (C. mukul), Acorus calamus (A. calamus) and Morshella esculenta (M. esculenta). Methanolic extraction of plants was carried out using maceration. Activation of yeast was performed using the sugar fermentation method. Pharmacological anti-aging screens in yeast performed by RLS. Primers for Actin (ACT1) and TOR1 were designed using Primer-BLAST. Gene expression analysis (RNA isolation and quantification, reverse transcription, and real-time polymerase chain reaction) of TOR1 in S. cerevisiae was conducted using methanolic extracts. Phenolic and Flavonoid compounds were quantified using reversed-phase high-performance liquid chromatography (RP-HPLC). Cell counting and viability assessment of S. cerevisiae with trypan blueusing methanolic extracts was performed. Gene expression analysis results revealed under expression of TOR1 in S. cerevisiae with mean normalized expression ratios < 1 using all four methanolic extracts. Results from RLS showed increased absorbances due to decreased TOR1 activity. HPLC analysis showed the presence of antioxidants such as vanillic acid, caffeic acid, rutin, and quercetin in crude extracts. Cell viability assessment revealed nontoxicity effects of M. esculenta and A. galangal whereas toxicity effects of A. calamus and C. mukul on S. cerevisiae cells. Reverse aging in S. cerevisiae was studied by gene expression analysis of TOR1 and RLS. TOR1 gene role in the activation of autophagy was explored using real-time polymerase chain reaction.