Objective: The present study was aimed to estimate the production of Mycophenolic acid by fungus Byssochlamys nivea strain JSR2 in various substrates and to conduct in silico studies to determine the interaction of MPA with the NS5B receptor of Hepatitis C Virus. Methods: The isolate was characterized as Byssochlamys nivea strain JSR2 by 18S rRNA gene sequencing. The production of mycophenolic acid by strain JSR2 was determined on various substrates such as pineapple and apple extract and was estimated using chromatographic technique. The morphological and spore arrangement of strain JSR2 was determined by high resolution scanning electron microscope. In silico studies were also conducted to determine the interaction of MPA with the NS5B receptor of hepatitis C virus with ribavarin as the reference ligand. Results: The mycophenolic acid production was found to be highest with pineapple juice substrate and was further identified by high performance liquid chromatography. The binding energy of MPA and ribavarin was found to be similar through Autodock Vina which further represents the same mechanism of action. Conclusion: The strain JSR2 produced good amount mycophenolic acid in the presence of pineapple juice as substrate and it represents the similar mode of action against Hepatitis C virus when compare to ribavaran with reference to in silico studies.