Table_2_Improving the Yield and Quality of Daptomycin in Streptomyces roseosporus by Multilevel Metabolic Engineering.DOCX

Daptomycin is a cyclic lipopeptide antibiotic with a significant antibacterial action against antibiotic-resistant Gram-positive bacteria. Despite numerous attempts to enhance daptomycin yield throughout the years, the production remains unsatisfactory. This study reports the application of multilevel metabolic engineering strategies in Streptomyces roseosporus to reconstruct high-quality daptomycin overproducing strain L2797-VHb, including precursor engineering (i.e., refactoring kynurenine pathway), regulatory pathway reconstruction (i.e., knocking out negative regulatory genes arpA and phaR), byproduct engineering (i.e., removing pigment), multicopy biosynthetic gene cluster (BGC), and fermentation process engineering (i.e., enhancing O2 supply). The daptomycin titer of L2797-VHb arrived at 113 mg/l with 565% higher comparing the starting strain L2790 (17 mg/l) in shake flasks and was further increased to 786 mg/l in 15 L fermenter. This multilevel metabolic engineering method not only effectively increases daptomycin production, but can also be applied to enhance antibiotic production in other industrial strains.

达托霉素（Daptomycin）是一种环状脂肽类抗生素，对耐药革兰氏阳性菌具有显著的抗菌活性。尽管多年来诸多研究尝试提升达托霉素的产量，但其工业化生产水平仍未达到理想状态。本研究报道了在玫瑰孢链霉菌（Streptomyces roseosporus）中应用多级代谢工程策略，重构得到优质达托霉素高产菌株L2797-VHb，所采用的策略包括：前体工程（即重构犬尿氨酸途径）、调控途径重构（即敲除负调控基因arpA与phaR）、副产物工程（即去除色素）、多拷贝生物合成基因簇（BGC）以及发酵工艺工程（即强化氧气供给）。在摇瓶培养条件下，菌株L2797-VHb的达托霉素效价达到113 mg/L，较出发菌株L2790（17 mg/L）提升565%；该效价在15 L发酵罐中进一步提升至786 mg/L。本多级代谢工程方法不仅可有效提升达托霉素的产量，还可应用于其他工业菌株的抗生素产量强化。