The　＂superbug＂　infection　caused　by　New　Delhi　metallo-beta-lactamase　(NDM-1)　has　become　an　emerging　threat.　Monitoring　NDM-1　has　proven　challenging　due　to　its　shuttling　between　pathogenic　bacteria.　Here,　we　report　an　isothermal　titration　calorimetry　(ITC)　method　that　can　monitor　activity　and　inhibition　of　NDM-1　in　live　bacterial　cells　in　real　time.　This　method　has　been　exemplified　by　monitoring　of　the　activity　and　inhibition　of　the　target　enzyme　and　evaluating　the　breakdown　of　antibiotics　by　pathogenic　bacteria　expressing　beta-lactamases.　Cell-based　studies　demonstrate　that　the　NDM-1　expressed　in　bacterial　cells　was　inhibited　by　four　known　inhibitors　ethylene　diamine　tetraacetic　acid　(EDTA),　D-captopril,　ebselen　and　azolylthioacetamide　with　fifty　percent　inhibitory　concentration　(IC50)　values　of　3.8,　48,　0.55,　and　17.5　mu　M,　respectively,　which　are　in　good　agreement　with　the　data　from　inhibition　kinetics　using　UV-vis　and　NMR　spectroscopy　in　vivo.　This　approach　could　be　applied　to　screen　and　evaluate　small　molecule　inhibitors　of　metallo-beta-lactamases　(M　beta　Ls)　in　whole　cells　or　to　identify　drug　resistant　bacteria.