Combinatorial biosynthesis of lipopeptide antibiotics in Streptomyces roseosporus
Combinatorial biosynthesis of lipopeptide antibiotics in Streptomyces roseosporus
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DOI：
10.1007/s10295-005-0030-y
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发表时间：
2006-02-01
影响因子：
3.4
通讯作者：
Wrigley, SK
中科院分区：
文献类型：
--
作者：
Baltz, RH;Brian, P;Wrigley, SK
达托霉素是一种由玫瑰链霉菌产生的环状脂肽抗生素。美比星((注册商标))(注射用达托霉素)于2003年被FDA批准用于治疗由革兰氏阳性病原体引起的皮肤和皮肤结构感染。达托霉素具有特别重要的意义，因为它代表了30年来第一个获准临床使用的新天然产品抗菌结构类别。达托霉素基因簇包含三个编码非核糖体肽合成酶(NRPS)的非常大的基因(dptA、dptBC和dptD)。相关的环脂肽A54145有四个NRPS基因(lptA、lptB、lptC和lptD)，而钙依赖抗生素(CDA)有三个(cdaPS1、cdaPS2和cdaPS3)。含有一个或多个NRPS基因缺失的玫瑰孢霉突变体，在ermEp*启动子的控制下，通过将这些基因插入到含有Phi C31或IS117连接(AttP Int)位点的接合克隆载体中，获得了dptA、dptBC和dptD的反式互补；通过与大肠杆菌的接合，将这些质粒定向插入到染色体中相应的attB位点。该反式互补系统被用来与lptD和cdaPS3产生亚基交换，重组子产生了新的杂化分子。多肽中D-丙氨酸(8)和D-丝氨酸(11)，I位的模块交换产生了额外的达托霉素的新衍生物。亚基交换和模块交换的方法与12位Glu的氨基酸修饰和脂类侧链起始单元的自然变异相结合，形成了与达托霉素相关的抗生素组合文库。许多工程菌株产生了适合分离和抗菌测试的新分子水平，而且大多数化合物都显示出抗菌活性。
Daptomycin is a cyclic lipopeptide antibiotic produced by Streptomyees roseosporus. Cubicin((R)) (daptomycin-for-injection) was approved in 2003 by the FDA to treat skin and skin structure infections caused by Gram-positive pathogens. Daptomycin is particularly significant in that it represents the first new natural product antibacterial structural class approved for clinical use in three decades. The daptomycin gene cluster contains three very large genes (dptA, dptBC, and dptD) that encode the nonribosomal peptide synthetase (NRPS). The related cyclic lipopeptide A54145 has four NRPS genes (lptA, lptB, lptC, and lptD), and calcium dependent antibiotic (CDA) has three (cdaPS1, cdaPS2, and cdaPS3). Mutants of S. roseosporus containing deletions of one or more of the NRPS genes have been trans-complemented with dptA, dptBC, and dptD by inserting these genes under the control of the ermEp* promoter into separate conjugal cloning vectors containing phi C31 or IS117 attachment (attP int) sites; delivering the plasmids into S. roseosporus by conjugation from Escherichia coli; and inserting the plasmids site-specifically into the chromosome at the corresponding attB sites. This trans-complementation system was used to generate subunit exchanges with lptD and cdaPS3 and the recombinants produced novel hybrid molecules. Module exchanges at positions D-Ala(8) and D-Ser(11), I in the peptide have produced additional novel derivatives of daptomycin. The approaches of subunit exchanges and module exchanges were combined with amino acid modifications of Glu at position 12 and natural variations in lipid side chain starter units to generate a combinatorial library of antibiotics related to daptomycin. Many of the engineered strains produced levels of novel molecules amenable to isolation and antimicrobial testing, and most of the compounds displayed antibacterial activities.