The endophytic fungus Periconia sp. produces piperine (5-(3, 4-methylenedioxyphenyl)-1-piperidinopent-2, 4-dien-1-one) under liquid culture. This is the first report of the alternative source for this chemical other than its host, Piper longum. The highly functionalized fungus-derived piperine exhibits strong antimycobacterial activity against Mycobacterium tuberculosis and M. smegmetis with minimum inhibitory concentrations of 1.74 and 2.62 μg ml(-1), respectively. The compound was crystallized and the structure was elucidated by single-crystal X-ray crystallography. This finding is of significance as piperine is a potential cancer preventative agent. It is reaffirmed by this report that important pharmaceuticals can be produced by endophytic microbes, and these molecules appear to be mimetic to their host origin. Therefore, we can enhance the bioactive principles of medicinal plants by isolating and identifying the endophytes, thereby showing the importance of preserving the biodiversity of these plants.