Other scientists, including Fleming, had tried to use penicillin prior to Paine, but with little evidence of success.
Paine shared his findings of successfully using penicillin to treat infection with Fleming and Sir Howard Florey, professor of pathology at Sheffield Medical School from 1932 to 1935.
Along with a team from Oxford, Florey then went on to purify penicillin, conducting the first clinical trial of the drug in 1941.
Paine did not publish his research, or maintain clear records, which meant his work could have been lost had it not been publicised in the 1980s.
Francis said: “We honour and commemorate his life and remarkable achievements in the city where he lived.
“Paine’s work is part of the University of Sheffield’s proud tradition of making discoveries that transform lives.”
The plaque commemorating Paine’s work has been installed at the University of Sheffield’s Jessop Building.