Lichens produce range of secondary metabolites which exhibit multifarious biological potential. The present work highlights the effect of media components for the production of secondary metabolites by lichen Parmotrema austrosinense. Mycobiont was isolated by lichen tissue culture method. One variable at a time approach exhibited that, Malt yeast extract media supplemented with glucose (4 mg/L), asparagine (1 mg/L), MnSO4 4H2O (20 mg/L), ZnSO4 7H2O (20 mg/L) and H3BO3 (4 mg/L) enhanced the growth of lichen mycobiont with improved total phenol production. The major influencing media components on total phenol content was screened through Plackett-Burman experimental design. Yeast, glucose and MgSO4 4H2O have been observed as effective media components. The screened variables have been further optimized by response surface methodology. The optimum media composition was found as 4.04 g/L glucose, 3.15 g/L yeast and 21.43 mg/L MnSO4 4H2O resulted with the maximum total phenol content of 1.80 ± 0.1 μg/g. Thus, the study established a suitable media component for the enhanced production rate of secondary metabolites in terms of total phenol content. Keywords: Parmotrema austrosinense; Secondary metabolites; Total phenol content; Optimization; Response surface; methodology.