Department of Biomedical Science and Engineering, Konkuk University, Seoul 05029, Republic of Korea.
J Am Chem Soc. 2024 Nov 6;146(44):30434-30442. doi: 10.1021/jacs.4c10966. Epub 2024 Oct 28.
Lipopeptides are an important family of natural products, some of which are clinically used as antibiotics to treat multidrug-resistant pathogens. Although the lipid moieties play a crucial role in balancing antibacterial activity and hemolytic toxicity, modifying the lipid moieties has been challenging due to the complexity of the lipidation process in lipopeptide biosynthesis. Here, we show that the lipid profile can be altered by engineering both secondary and primary metabolisms, using daptomycin as an example. First, swapping the fatty acyl AMP ligase (FAAL) gene with foreign FAAL homologs improved the fatty acyl specificity of the lipidation process for decanoic acid. Then, the introduction of type I fatty acid synthase operon (MvFAS-Ib/MvAcpS) and thioesterase (CpTEII) enriched the fatty acid pool with decanoic acid in . The engineered fatty acid metabolism eliminates the need for external decanoic acid supplementation by enabling to biosynthesize decanoic acid. By complete engineering of the lipidation process, we achieved, for the first time, high-purity, natural production of daptomycin. The lipidation engineering approach we demonstrate here lays the foundation for the lipidation control in lipopeptide biosynthesis.
脂肽是一类重要的天然产物，其中一些被临床用作抗生素来治疗多药耐药病原体。尽管脂质部分在平衡抗菌活性和溶血毒性方面起着至关重要的作用，但由于脂肽生物合成中脂质化过程的复杂性，修饰脂质部分一直具有挑战性。在这里，我们以达托霉素为例，展示了通过工程化二次和初级代谢来改变脂质谱的方法。首先，用外源 FAAL 同源物替换脂肪酸 AMP 连接酶 (FAAL) 基因，改善了癸酸的脂化过程中的脂肪酸酰基特异性。然后，引入 I 型脂肪酸合酶操纵子 (MvFAS-Ib/MvAcpS) 和硫酯酶 (CpTEII)，使脂肪酸库中癸酸丰富。工程化的脂肪酸代谢通过使 能够生物合成癸酸，消除了对外源癸酸补充的需求。通过完全工程化的脂化过程，我们首次实现了达托霉素的高纯度、天然生产。我们在这里展示的脂化工程方法为脂肽生物合成中的脂化控制奠定了基础。
