Miao Vivian, Coëffet-Le Gal Marie-Francois, Nguyen Kien, Brian Paul, Penn Julia, Whiting Andrew, Steele Jonathan, Kau David, Martin Steve, Ford Robert, Gibson Trevor, Bouchard Mario, Wrigley Stephen K, Baltz Richard H
Cubist Pharmaceuticals, Inc., 65 Hayden Avenue, Lexington, Massachusetts 02421, USA.
Chem Biol. 2006 Mar;13(3):269-76. doi: 10.1016/j.chembiol.2005.12.012.
Daptomycin is a lipopeptide antibiotic produced by a nonribosomal peptide synthetase (NRPS) in Streptomyces roseosporus. The holoenzyme is composed of three subunits, encoded by the dptA, dptBC, and dptD genes, each responsible for incorporating particular amino acids into the peptide. We introduced expression plasmids carrying dptD or NRPS genes encoding subunits from two related lipopeptide biosynthetic pathways into a daptomycin nonproducing strain of S. roseosporus harboring a deletion of dptD. All constructs successfully complemented the deletion in trans, generating three peptide cores related to daptomycin. When these were coupled with incomplete methylation of 1 amino acid and natural variation in the lipid side chain, 18 lipopeptides were generated. Substantial amounts of nine of these compounds were readily obtained by fermentation, and all displayed antibacterial activity against gram-positive pathogens.
达托霉素是由玫瑰孢链霉菌中的非核糖体肽合成酶（NRPS）产生的一种脂肽抗生素。全酶由三个亚基组成，分别由dptA、dptBC和dptD基因编码，每个亚基负责将特定氨基酸掺入肽中。我们将携带dptD或来自两个相关脂肽生物合成途径的亚基编码NRPS基因的表达质粒导入缺失dptD的达托霉素不产生产株玫瑰孢链霉菌中。所有构建体均成功地在反式中互补缺失，产生了三种与达托霉素相关的肽核心。当这些肽核心与1个氨基酸的不完全甲基化和脂质侧链的自然变异相结合时，产生了18种脂肽。通过发酵很容易获得大量的其中9种化合物，并且所有这些化合物都对革兰氏阳性病原体显示出抗菌活性。
