TAILIEUCHUNG - Báo cáo khoa học: Biochemical characterization and inhibitor discovery of shikimate dehydrogenase from Helicobacter pylori

Shikimate dehydrogenase (SDH) is the fourth enzyme involved in the shiki-mate pathway. It catalyzes the NADPH-dependent reduction of 3-dehy-droshikimate to shikimate, and has been developed as a promising target for the discovery of antimicrobial agent. In this report, we identified a new aroEgene encoding SDH fromHelicobacter pyloristrain SS1. The recom-binantH. | ễFEBS Journal Biochemical characterization and inhibitor discovery of shikimate dehydrogenase from Helicobacter pylori Cong Han1 Lirui Wang1 Kunqian Yu1 Lili Chen1 Lihong Hu1 Kaixian Chen1 Hualiang Jiang1 2 and Xu Shen1 2 1 Drug Discovery and Design Center State Key Laboratory of Drug Research Shanghai Institute of Materia Medica Chinese Academy of Sciences Shanghai China 2 Schoolof Pharmacy East China University of Science and Technology Shanghai China Keywords antibacterial agent drug target enzyme inhibition Helicobacter pylori shikimate dehydrogenase Correspondence X. Shen H. Jiang and L. Hu Shanghai Institute of Materia Medica Chinese Academy of Sciences 555 Zu Chong Zhi Road Zhangjiang Hi-Tech Park Shanghai 201203 China. Tel Fax 86 21 50806918 E-mail xshen@ hjiang@ simmkulh@ Database The sequence reported in this paper has been submitted to GenBank database under accession number AY738333 Received 23 April2006 revised 11 July 2006 accepted 16 August 2006 doi Shikimate dehydrogenase SDH is the fourth enzyme involved in the shiki-mate pathway. It catalyzes the NADPH-dependent reduction of 3-dehy-droshikimate to shikimate and has been developed as a promising target for the discovery of antimicrobial agent. In this report we identified a new aroE gene encoding SDH from Helicobacter pylori strain SS1. The recombinant H. pylori shikimate dehydrogenase HpSDH was cloned expressed and purified in Escherichia coli system. The enzymatic characterization of HpSDH demonstrates its activity with kcat of s-1 and Km of mM toward shikimate kcat of s-1 and Km of mM toward NADP kcat of s-1 and Km of mM toward NAD. The optimum pH of the enzyme activity is between and and the optimum temperature is around 60 C. Using high throughput screening against our laboratory chemical library five compounds curcumin 1 3- 2-naphthyloxy -4-oxo-2- trifluoromethyl .

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