The enzymes involved in bacterial cell wall synthesis are established antibiotic targets, and continue to be a central focus for antibiotic development. Bacterial penicillin-binding proteins (and, in some bacteria, l,d-transpeptidases) form essential peptide cross-links in the cell wall. Although the β-lactam class of antibiotics target these enzymes, bacterial resistance threatens their clinical use, and there is an urgent unmet need for new antibiotics. However, the search for new antibiotics targeting the bacterial cell wall is hindered by a number of obstacles associated with screening the enzymes involved in peptidoglycan synthesis. This review describes recent approaches for measuring the activity and inhibition of penicillin-binding proteins and l,d-transpeptidases, highlighting strategies that are poised to serve as valuable tools for high-throughput screening of transpeptidase inhibitors, supporting the development of new antibiotics.

细菌细胞壁合成中涉及的酶是确定的抗生素靶标，并且继续是抗生素开发的中心焦点。细菌青霉素结合蛋白（在某些细菌中是l，d- transpeptidases）在细胞壁上形成必需的肽交联。尽管β-内酰胺类抗生素以这些酶为靶标，但细菌耐药性威胁了它们的临床应用，并且对新抗生素的迫切需求未得到满足。然而，与筛选涉及肽聚糖合成的酶有关的许多障碍阻碍了寻找靶向细菌细胞壁的新抗生素。这篇综述描述了测量青霉素结合蛋白和l，d的活性和抑制作用的最新方法-转肽酶，突出显示准备用作高通量筛选转肽酶抑制剂的有价值工具的策略，支持开发新的抗生素。