College of Biological Science and Technology, Fuzhou University, Fujian, China.
Lett Appl Microbiol. 2012 Jul;55(1):33-9. doi: 10.1111/j.1472-765X.2012.03254.x. Epub 2012 May 18.
To engineer Streptomyces tenebrarius for producing carbamoyl tobramycin as a main component.
The aprH-M gene fragment (apramycin biosynthetic gene from GenBank) in S. tenebrarius Tt49 was knocked out by genetic engineering to form S. tenebrarius T106 (ΔaprH-M). Compared to the wild-type strain, mutant strain T106 (ΔaprH-M) no longer produced apramycin, while mainly synthesize carbamoyl tobramycin. TLC and HPLC-MS analyses indicated that the mutant strain significantly increased the production of carbamoyl tobramycin.
The metabolic flow for the apramycin and its analogues biosynthesis was blocked by disrupting the aprH-M gene clusters. The aprH-M gene clusters might be essential for the biosynthesis of apramycin. The mutant strain T106 mainly synthesized carbamoyl tobramycin.
The mutant T106 mainly produces carbamoyl tobramycin without synthesizing apramycin, which will reduce cost of postextraction from fermentation products. Therefore, it has good prospects for industrial application.
通过基因工程改造暗灰色链霉菌以生产氨甲酰妥布霉素作为主要成分。
通过基因工程敲除暗灰色链霉菌 Tt49 中的 aprH-M 基因片段（来自 GenBank 的安普霉素生物合成基因），形成暗灰色链霉菌 T106（ΔaprH-M）。与野生型菌株相比，突变株 T106（ΔaprH-M）不再产生安普霉素，而主要合成氨甲酰妥布霉素。TLC 和 HPLC-MS 分析表明，突变株显著增加了氨甲酰妥布霉素的产量。
通过破坏 aprH-M 基因簇，阻断了安普霉素及其类似物生物合成的代谢流。aprH-M 基因簇可能是安普霉素生物合成所必需的。突变株 T106 主要合成氨甲酰妥布霉素。
突变株 T106 主要产生氨甲酰妥布霉素而不合成安普霉素，这将降低从发酵产物中提取的成本。因此，它具有良好的工业应用前景。
