Deoxysugars in glycopeptide antibiotics: Enzymatic synthesis of TDP-L-epivancosamine in chloroeremomycin biosynthesis
Deoxysugars in glycopeptide antibiotics: Enzymatic synthesis of TDP-L-epivancosamine in chloroeremomycin biosynthesis
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DOI：
10.1073/pnas.210395097
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发表时间：
2000-10-24
影响因子：
11.1
通讯作者：
Burkart, MD
中科院分区：
文献类型：
--
作者：
Chen, HW;Thomas, MG;Burkart, MD
2，3，6-三脱氧糖L-表万古胺是添加到氯伊瑞霉素中的糖苷配基支架的末端糖。糖肽抗生素中万古霉素家族的一员。来自氯伊瑞霉素生物合成簇的五种蛋白质，ORF 14和ORF 23至ORF 26。已在大肠杆菌中异源表达并纯化至接近同质，并且每种都已表征了酶活性。这五种酶重建了从TDP-4-酮-6-脱氧-D-葡萄糖的TDP-L-表万古胺的完全生物合成。该过程涉及C-2脱氧、C-3胺化和甲基化、C-5差向异构化和C-4酮还原。该途径的中间体和最终产物已通过质谱和NMR鉴定。这里建立的途径代表了一个不寻常的糖的一个重要类别的抗生素的完整的体外重建，并为未来的组合生物合成新的生物活性化合物奠定了基础。
The 2,3,6-trideoxysugar L-epivancosamine is the terminal sugar added to the aglycone scaffold in chloroeremomycin. a member of the vancomycin family of glycopeptide antibiotics. Five proteins from the chloroeremomycin biosynthetic cluster, ORF14 and ORF23 to ORF26. have been expressed heterologously in Escherichia coli and purified to near homogeneity, and each has been characterized for an enzymatic activity. These five enzymes reconstitute the complete biosynthesis of TDP-L-epivancosamine from TDP-4-keto-6-deoxy-D-glucose. This process involves C-2 deoxygenation, C-3 amination and methylation, C-5 epimerization, and C-4 ketoreduction. Intermediates and the final product of this pathway have been identified by mass spectrometry and NMR. The pathway established here represents the complete in vitro reconstitution of an unusual sugar for an important class of antibiotics and sets the groundwork for future combinatorial biosynthesis for new bioactive compounds.