金属β-内酰胺酶（MβLs）包括NDM-1所导致的“超级细菌”耐药是人们目前密切关注的热点。该项目拟表达并纯化所有三个B组群具代表性的MβLs，包括 NDM-1、CcrA、ImiS及L1等；构建和发展唑基硫代乙酰胺衍生物，表征其化学组成与结构，酶动力学测定其对MβLs的抑制活性；MIC评价其协同抗生素对产MβLs耐药菌的抑制效能， ITC测定其抑制MβLs水解抗生素的热力学函数及酶稳定构象态的转化；重组Co-MβLs，光谱表征抑制剂与MβLs的作用及作用强度、抑制剂的键合位点及键合方向等；浸透法制备抑制剂-MβLs复合物之有序晶体，解析结构获得其作用模式的直接证据；创新一种对细胞非侵入性的UV-Vis方法，实时监测耐药菌活细胞体内MβLs的活性，进行原位抑制研究，进而探索在复杂生物网络环境中MβLs的结构以及与其它细胞内成分的互动等。该项目的实施，将为发展新型抗菌药物提供有重要价值的信息。

Metallo-β-lactamase (MβLs) including NDM-1 led to the "super bacteria" resistance, which is the hot spot that people currently pay close attention to. The project intends to do expression and purification of the MβLs from each B subclass of them, inclusding NDM-1, CcrA, ImiS and L1 etc.; construction and development of azolyl thioacetamide derivatives, characterization of their chemical composition and structures, evaluation of their inhibitory activity against MβLs by enzyme kinetics; testing their synergies in combination with antibiotics to inhibit the antibiotic resistant bacteria with MIC method; determination of the thermodynamic parameters of them inhibiting MβLs and conversion of enzyme stability conformation states using isothermy titration microcalorimetry (ITC); recombinant of Co (II) substituted Co-MβLs, spectroscopy characterization of interaction between inhibitors and MβLs and effect of intensity, bonding sites and bonding direction of the inhibitor etc.; preparation of the ordered crystals of inhibitor-MβLs complexes by soaking technology, and obtaining direct evidence by resolution of the interaction mode; using an innovation UV-Vis method of cell non-invasive, real-time monitoring the activity of MβLs in living cells of antibiotic resistant bacteria, and studing on inhibition of the enzymes in situ, further exploration of the structure of MβLs and their interaction with other cellular components in complex biological network environment etc. Run of this project will provide information with important valuable for the development of new antibacterial drugs.