Nguyen Kien T, Ritz Daniel, Gu Jian-Qiao, Alexander Dylan, Chu Min, Miao Vivian, Brian Paul, Baltz Richard H
Department of Drug Discovery and Evaluation, Cubist Pharmaceuticals, 65 Hayden Avenue, Lexington, MA 02421, USA.
Proc Natl Acad Sci U S A. 2006 Nov 14;103(46):17462-7. doi: 10.1073/pnas.0608589103. Epub 2006 Nov 7.
Daptomycin, a cyclic lipopeptide produced by Streptomyces roseosporus, is the active ingredient of Cubicin (daptomycin-for-injection), a first-in-class antibiotic approved for treatment of skin and skin-structure infections caused by Gram-positive pathogens and bacteremia and endocarditis caused by Staphylococcus aureus, including methicillin-resistant strains. Genetic engineering of the nonribosomal peptide synthetase (NRPS) in the daptomycin biosynthetic pathway was exploited for the biosynthesis of novel active antibiotics. lambda-Red-mediated recombination was used to exchange single or multiple modules in the DptBC subunit of the NRPS to modify the daptomycin cyclic peptide core. We combined module exchanges, NRPS subunit exchanges, inactivation of the tailoring enzyme glutamic acid 3-methyltransferase, and natural variations of the lipid tail to generate a library of novel lipopeptides, some of which were as active as daptomycin against Gram-positive bacteria. One compound was more potent against an Escherichia coli imp mutant that has increased outer membrane permeability. This study established a robust combinatorial biosynthesis platform to produce novel peptide antibiotics in sufficient quantities for antimicrobial screening and drug development.
达托霉素是由玫瑰孢链霉菌产生的一种环脂肽，是 Cubicin（注射用达托霉素）的活性成分，Cubicin 是一种一流的抗生素，被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以及由金黄色葡萄球菌引起的菌血症和心内膜炎，包括耐甲氧西林菌株。达托霉素生物合成途径中非核糖体肽合成酶（NRPS）的基因工程被用于新型活性抗生素的生物合成。λ-Red 介导的重组用于交换 NRPS 的 DptBC 亚基中的单个或多个模块，以修饰达托霉素环肽核心。我们结合模块交换、NRPS 亚基交换、修饰酶谷氨酸 3-甲基转移酶的失活以及脂质尾部的自然变异，生成了一个新型脂肽文库，其中一些脂肽对革兰氏阳性菌的活性与达托霉素相当。有一种化合物对具有增加的外膜通透性的大肠杆菌 imp 突变体更有效。这项研究建立了一个强大的组合生物合成平台，以生产足够数量的新型肽抗生素用于抗菌筛选和药物开发。
