CmlI是氯霉素生物合成中的一种加氧酶。
CmlI is an -oxygenase in the biosynthesis of chloramphenicol.
作者信息
Lu Haige, Chanco Emmanuel, Zhao Huimin
机构信息
Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, Urbana, IL 61801, USA.
Department of Chemistry, University of Illinois at Urbana-Champaign, USA.
出版信息
Tetrahedron. 2012 Sep 16;68(37). doi: 10.1016/j.tet.2012.06.036.
Abstract
The N-oxygenation of an amine group is one of the steps in the biosynthesis of the antibiotic chloramphenicol. The non-heme di-iron enzyme CmlI was identified as the enzyme catalyzing this reaction through bioinformatics studies and reconstitution of enzymatic activity. In vitro reconstitution was achieved using phenazine methosulfate and NADH as electron mediators, while in vivo activity was demonstrated in using two substrates. Kinetic analysis showed a biphasic behavior of the enzyme. Oxidized hydroxylamine and nitroso compounds in the reaction were detected both in vitro and in vivo based on LC-MS. The active site metal was confirmed to be iron based on a ferrozine assay. These findings provide new insights into the biosynthesis of chloramphenicol and could lead to further development of CmlI as a useful biocatalyst.
摘要
胺基的N-氧化是抗生素氯霉素生物合成过程中的步骤之一。通过生物信息学研究和酶活性重建，非血红素双铁酶CmlI被鉴定为催化该反应的酶。使用硫酸吩嗪和NADH作为电子介质实现了体外重建，而在体内使用两种底物证明了其活性。动力学分析表明该酶具有双相行为。基于LC-MS在体外和体内均检测到反应中的氧化羟胺和亚硝基化合物。基于亚铁嗪测定法确认活性位点金属为铁。这些发现为氯霉素的生物合成提供了新的见解，并可能导致CmlI作为一种有用的生物催化剂得到进一步开发。
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