Mineral trioxide aggregate (MTA) is a relatively new versatile dental material. MTA has many advantages as well as disadvantages. To reduce most of the drawbacks of MTA, a premixed bioceramic MTA, NeoPutty MTA, was introduced in 2020. In this study, we assessed the antimicrobial activity of the newer MTA, NeoPutty MTA. We modified NeoPutty MTA and compared both against Enterococcus faecalis, Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli. Using the agar diffusion method, NeoPutty MTA was tested for antibacterial activity against the above-mentioned microorganisms. A base layer of Petri plates was done using Muller-Hinton agar for S. aureus, E. coli, and P. aeruginosa and brain heart infusion agar for E. faecalis. A total of 32 plates were employed; the plates were divided randomly into four test groups having eight plates each, so microorganisms were tested eight times. Three cavities were made in agar and filled with freshly mixed materials after 24 h. A pour plate seeded the microorganisms. The plates were pre-incubated for 2 h at room temperature and incubated at 37°C for 24 h. An independent observer measured the inhibition zone diameters. NeoPutty MTA, when tested alone, did not show much antibacterial activity against E. faecalis, S. aureus, and E. coli but had significant antimicrobial activity against P. aeruginosa when used at different concentrations. Modified NeoPutty (NeoPutty with antibiotics added individually) showed significant antibacterial activity against these microorganisms, as seen by the zone of inhibition of these bacteria. Modified NeoPutty with antibiotics has a better antimicrobial effect than NeoPutty MTA.