Synthesis and derivatization of daptomycin:: A chemoenzymatic route to acidic lipopeptide antibiotics
Synthesis and derivatization of daptomycin:: A chemoenzymatic route to acidic lipopeptide antibiotics
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DOI：
10.1021/ja045455t
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发表时间：
2004-12-29
影响因子：
15
通讯作者：
Marahiel, MA
中科院分区：
文献类型：
--
作者：
Grünewald, J;Sieber, SA;Marahiel, MA
达托霉素是一种支链环状非核糖体组装的酸性脂肽，是第一个临床批准的该类抗生素。在这里，我们表明，天蓝色链霉菌钙依赖性抗生素（CDA）的非核糖体肽合成酶（NRPS）的重组环化结构域是一个通用的工具，为达托霉素衍生物的化学酶代。在6个达托霉素特异性残基处具有单交换的线性CDA十一肽硫酯被CDA环化酶成功地环化。将所有6个残基同时掺入肽骨架中并将CDA的N-末端延长2个残基，得到仅缺少L-3-甲基谷氨酸盐的β-甲基的达托霉素衍生物。几种底物类似物的生物活性研究揭示了非蛋白质成分对抗菌效力的重要作用。与酸性脂肽一致。化学酶促组装的达托霉素类似物的生物活性取决于油钙离子的浓度。肽骨架中四个酸性残基的单一缺失表明只有两个天冬氨酸残基对于抗微生物效力是必需的。这两个残基在其他非核糖体酸性脂肽和核糖体组装的钙调蛋白的EF基序中是严格保守的。基于这些发现，CDA环化酶是一种通用催化剂，其可用于产生新的达托霉素衍生物，所述衍生物难以通过母体十三肽的化学修饰来获得以进一步改善其治疗活性。
Daptomycin is a branched cyclic nonribosomally assembled acidic lipopeptide, which is the first clinically approved antibiotic of this class. Here we show that the recombinant cyclization domain of the Streptomyces coelicolor calcium-dependent antibiotic (CDA) nonribosomal peptide synthetase (NRPS) is a versatile tool for the chemoenzymatic generation of daptomycin derivatives. Linear CDA undecapeptide thioesters with single exchanges at six daptomycin-specific residues were successfully cyclized by CDA cyclase. Simultaneous incorporation of all six of these residues into the peptide backbone and elongation of the N-terminus of CDA by two residues yielded a daptomycin derivative that lacked only the beta-methyl group of L-3-methylglutamate. Bioactivity studies with several substrate analogues revealed a significant role of nonproteinogenic constituents for antibacterial potency. In accordance with acidic lipopeptides. the bioactivity of the chemoenzymatic assembled daptomycin analogue is dependent on the Concentration oil calcium ions. Single deletions of the four acidic residues in the peptide backbone suggest that only two aspartic acid residues are essential for antimicrobial potency. These two residues are strictly conserved among other nonribosomal acidic lipopeptides and the EF-motif of ribosomally assembled calmodulin. Based on these findings CDA cyclase is a versatile catalyst that can be used to generate novel daptomycin derivatives that are otherwise difficult to obtain by chemical modification of the parental tridecapeptide to improve further its therapeutic activity.