R. factors conferring resistance to penicillins and cephalosporins were transferred by conjugation from six strains of Shigella and from four strains of Salmonella typhimurium to a standard strain of S. typhimurium LT2. The beta-lactamases produced were then characterized by using cell-free extracts. The enzymes were of two types, I and II, with respect to specific activity against benzyl penicillin, substrate profile, K(m), pH optimum, temperature optimum, inhibition by chloride and nitrate ions, and heat inactivation. The six type I enzymes were associated with R factors from Shigella strains; five of these were R(f) factors. The four type II enzymes were associated with R factors from S. typhimurium; all these were R(i) factors.