Stassi D, Post D, Satter M, Jackson M, Maine G
Pharmaceutical Products Division, Abbott Laboratories, Abbott Park, 1401 Sheridan Road, North Chicago, IL 60064, USA.
Appl Microbiol Biotechnol. 1998 Jun;49(6):725-31. doi: 10.1007/s002530051238.
The erythromycin producer, Saccharopolyspora erythraea ER720, was genetically engineered to produce 6,12-dideoxyerythromycin A, a novel erythromycin derivative, as the major macrolide in the fermentation broth. Inspection of the biosynthetic pathway for erythromycin would suggest that production of this compound could be achieved simply through the disruption of two genes, that encoding the erythromycin C-6 hydroxylase (eryF) and that encoding the erythromycin C-12 hydroxylase (eryK). The double mutant, however, was found to produce a mixture of 6,12-dideoxyerythromycin A and the precursor, 6-deoxyerythromycin D. Complete conversion to the desired product (to the limit of detection by TLC) was achieved by inserting an additional copy of the eryG gene, encoding the erythromycin 3"-O-methyltransferase and driven by the ermE* promoter, into the S. erythraea chromosome.
对红霉素产生菌糖多孢红霉菌ER720进行基因工程改造，使其产生一种新型红霉素衍生物6,12-二脱氧红霉素A作为发酵液中的主要大环内酯类化合物。对红霉素生物合成途径的研究表明，只需破坏两个基因，即编码红霉素C-6羟化酶的基因（eryF）和编码红霉素C-12羟化酶的基因（eryK），就可以实现该化合物的生产。然而，发现双突变体产生的是6,12-二脱氧红霉素A和前体6-脱氧红霉素D的混合物。通过将编码红霉素3"-O-甲基转移酶且由ermE*启动子驱动的eryG基因的额外拷贝插入糖多孢红霉菌染色体，实现了完全转化为所需产物（达到TLC检测极限）。
