当前位置:
X-MOL 学术
›
Metab. Eng.
›
论文详情
Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
Combinatorial metabolic engineering of Bacillus subtilis for de novo production of polymyxin B
Metabolic Engineering ( IF 6.8 ) Pub Date : 2024-04-04 , DOI: 10.1016/j.ymben.2024.04.001 Hui-Zhong Sun 1 , Qing Li 1 , Wei Shang 1 , Bin Qiao 1 , Qiu-Man Xu 2 , Jing-Sheng Cheng 1
Metabolic Engineering ( IF 6.8 ) Pub Date : 2024-04-04 , DOI: 10.1016/j.ymben.2024.04.001 Hui-Zhong Sun 1 , Qing Li 1 , Wei Shang 1 , Bin Qiao 1 , Qiu-Man Xu 2 , Jing-Sheng Cheng 1
Affiliation
Polymyxin is a lipopeptide antibiotic that is effective against multidrug-resistant Gram-negative bacteria. However, its clinical development is limited due to low titer and the presence of homologs. To address this, the polymyxin gene cluster was integrated into Bacillus subtilis , and sfp from Paenibacillus polymyxa was expressed heterologously, enabling recombinant B. subtilis to synthesize polymyxin B. Regulating NRPS domain inhibited formation of polymyxin B2 and B3. The production of polymyxin B increased to 329.7 mg/L by replacing the native promoters of pmxA , pmxB , and pmxE with PfusA , C2up, and PfusA , respectively. Further enhancement in this production, up to 616.1 mg/L, was achieved by improving the synthesis ability of 6-methyloctanoic acid compared to the original strain expressing polymyxin heterologously. Additionally, incorporating an anikasin-derived domain into the hybrid nonribosomal peptide synthase of polymyxin increased the B1 ratio in polymyxin B from 57.5% to 62.2%. Through optimization of peptone supply in the fermentation medium and fermentation in a 5.0-L bioreactor, the final polymyxin B titer reached 962.1 mg/L, with a yield of 19.24 mg/g maltodextrin and a productivity of 10.02 mg/(L·h). This study demonstrates a successful approach for enhancing polymyxin B production and increasing the B1 ratio through combinatorial metabolic engineering.
中文翻译:
枯草芽孢杆菌的组合代谢工程用于从头生产多粘菌素 B
多粘菌素是一种脂肽抗生素,对多重耐药革兰氏阴性菌有效。然而,由于低滴度和同源物的存在,其临床开发受到限制。为了解决这个问题,将多粘菌素基因簇整合到枯草芽孢杆菌中,并且来自 Paenibacillus polymyxa 的 sfp 异源表达,使重组枯草芽孢杆菌能够合成多粘菌素 B。通过分别用 PfusA、C2up 和 PfusA 取代 pmxA 、 pmxB 和 pmxE 的天然启动子,多粘菌素 B 的产量增加到 329.7 mg/L。与异源表达多粘菌素的原始菌株相比,通过提高 6-甲基辛酸的合成能力,实现了该产量的进一步增强,高达 616.1 mg/L。此外,将异香蛋白衍生结构域掺入多粘菌素的杂化非核糖体肽合酶中,可将多粘菌素 B 中的 B1 比率从 57.5% 提高到 62.2%。通过优化发酵培养基中的蛋白胨供应,并在 5.0 L 生物反应器中发酵,多粘菌素 B 最终滴度达到 962.1 mg/L,麦芽糖糊精产量为 19.24 mg/g,产能为 10.02 mg/(L·h)。本研究展示了一种通过组合代谢工程增强多粘菌素 B 产生和增加 B1 比率的成功方法。
更新日期:2024-04-04
中文翻译:
枯草芽孢杆菌的组合代谢工程用于从头生产多粘菌素 B
多粘菌素是一种脂肽抗生素,对多重耐药革兰氏阴性菌有效。然而,由于低滴度和同源物的存在,其临床开发受到限制。为了解决这个问题,将多粘菌素基因簇整合到枯草芽孢杆菌中,并且来自 Paenibacillus polymyxa 的 sfp 异源表达,使重组枯草芽孢杆菌能够合成多粘菌素 B。通过分别用 PfusA、C2up 和 PfusA 取代 pmxA 、 pmxB 和 pmxE 的天然启动子,多粘菌素 B 的产量增加到 329.7 mg/L。与异源表达多粘菌素的原始菌株相比,通过提高 6-甲基辛酸的合成能力,实现了该产量的进一步增强,高达 616.1 mg/L。此外,将异香蛋白衍生结构域掺入多粘菌素的杂化非核糖体肽合酶中,可将多粘菌素 B 中的 B1 比率从 57.5% 提高到 62.2%。通过优化发酵培养基中的蛋白胨供应,并在 5.0 L 生物反应器中发酵,多粘菌素 B 最终滴度达到 962.1 mg/L,麦芽糖糊精产量为 19.24 mg/g,产能为 10.02 mg/(L·h)。本研究展示了一种通过组合代谢工程增强多粘菌素 B 产生和增加 B1 比率的成功方法。