玫瑰黄素高产微生物的代谢工程。

Metabolic engineering of roseoflavin-overproducing microorganisms.

机构信息

Institute for Technical Microbiology, Mannheim University of Applied Sciences, Paul-Wittsack-Str. 10, 68163, Mannheim, Germany.

出版信息

Microb Cell Fact. 2019 Aug 26;18(1):146. doi: 10.1186/s12934-019-1181-2.

BACKGROUND

Roseoflavin, a promising broad-spectrum antibiotic, is naturally produced by the bacteria Streptomyces davaonensis and Streptomyces cinnabarinus. The key enzymes responsible for roseoflavin biosynthesis and the corresponding genes were recently identified. In this study we aimed to enhance roseoflavin production in S. davaonensis and to synthesize roseoflavin in the heterologous hosts Bacillus subtilis and Corynebacterium glutamicum by (over)expression of the roseoflavin biosynthesis genes.

RESULTS

While expression of the roseoflavin biosynthesis genes from S. davaonensis was not observed in recombinant strains of B. subtilis, overexpression was successful in C. glutamicum and S. davaonensis. Under the culture conditions tested, a maximum of 1.6 ± 0.2 µM (ca. 0.7 mg/l) and 34.9 ± 5.2 µM (ca. 14 mg/l) roseoflavin was produced with recombinant strains of C. glutamicum and S. davaonensis, respectively. In S. davaonensis the roseoflavin yield was increased by 78%.

CONCLUSIONS

The results of this study provide a sound basis for the development of an economical roseoflavin production process.

背景

玫瑰黄素是一种有前途的广谱抗生素，天然由细菌地衣芽孢杆菌和绛红小单孢菌产生。负责玫瑰黄素生物合成的关键酶和相应的基因最近已被确定。在这项研究中，我们旨在通过（过度）表达玫瑰黄素生物合成基因来提高地衣芽孢杆菌中玫瑰黄素的产量，并在异源宿主枯草芽孢杆菌和谷氨酸棒杆菌中合成玫瑰黄素。

结果

虽然在枯草芽孢杆菌的重组菌株中没有观察到来自地衣芽孢杆菌的玫瑰黄素生物合成基因的表达，但在谷氨酸棒杆菌和地衣芽孢杆菌中成功地进行了过表达。在测试的培养条件下，重组谷氨酸棒杆菌和地衣芽孢杆菌菌株分别产生了 1.6±0.2µM（约 0.7mg/l）和 34.9±5.2µM（约 14mg/l）的最大玫瑰黄素量。在地衣芽孢杆菌中，玫瑰黄素的产量增加了 78%。