Mitsakos Voula
School of Chemistry, The University of Melbourne, Parkville, VIC 3010, Australia.
Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Parkville, VIC 3010, Australia.
Heliyon. 2023 Mar 6;9(3):e14304. doi: 10.1016/j.heliyon.2023.e14304. eCollection 2023 Mar.
In search of a new class of antibacterial agents, compounds that target the essential bacterial enzyme, dihydrodipicolinate synthase (DHDPS), are of interest to drug discovery efforts. DHDPS catalyzes the first committed step in the diaminopimelate (DAP) pathway to the biosynthesis of lysine in bacteria and plants. The -aminobenzaldehyde (-ABA) assay is typically used as a qualitative tool for identifying fractions containing DHDPS during purification. This report is about the development of a high-throughput -ABA assay format for the quantification of DHDPS enzyme activity using multi-well plates. The colorimetric assay is suitable for determining enzymatic parameters ( and V) and identifying inhibitors of DHDPS in a high-throughput screen.
为寻找新型抗菌剂,能够作用于关键细菌酶——二氢二吡啶酸合酶(DHDPS)的化合物成为药物研发工作关注的对象。DHDPS催化细菌和植物中赖氨酸生物合成二氨基庚二酸(DAP)途径中的首个关键步骤。在纯化过程中,α-氨基苯甲醛(α-ABA)测定法通常用作鉴定含有DHDPS组分的定性工具。本报告介绍了一种高通量α-ABA测定法的开发,该方法使用多孔板对DHDPS酶活性进行定量。这种比色测定法适用于在高通量筛选中确定酶促参数(Km和Vmax)并鉴定DHDPS抑制剂。