The non-edible mushrooms of Ganoderma lucidum, Fomes fomentarius and Ganoderma tsugae were collected and studied for their antibacterial activity against the pathogenic bacteria such as Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumoniae by using well diffusion method. The acetone extracts of Ganoderma lucidum had highest zone of inhibition against Pseudomonas aeruginosa. The minimum zone of inhibition was found in Ganoderma tsugae against Klebsiella pneumoniae and Fomes fomentarius against Klebsiella pneumoniae and Bacillus subtilis. In ethanol extracts, the highest zone of inhibition was found in Fomes fomentarius against all the tested microorganisms followed by Ganoderma lucidum and Ganoderma tsugae. It was noted that the Fomes fomentarius extract have superior antibacterial activity than the other mushroom extracts. The gram negative bacteria Pseudomonas aeruginosa was more susceptible than other tested microorganisms. The alkaloid content of the mushroom powder (Ganoderma lucidum, Fomes fomentarius and Ganoderma tsugae) was analysed by using HPTLC method. As per the results Ganoderma lucidum reported the presence of one kind of alkaloid and in Ganoderma tsugae the presence of two kinds of alkaloids was found in UV 254nm. The alkaloid content was absent in Fomes fomentarius.