中文摘要：

plasmid 表示系统被潜在的 plasmid 不稳定性习惯性地折磨。Chromosomal 集成是一条强大的途径克服这个问题。此处，我们报导 plasmid 免费的超 E。为辅酶 Q10 生产的 coli 紧张。一系列集成表示向量， pxKC3T5b 和 pxKT5b，分别地为化学上可诱导的 chromosomal 进化(多重拷贝集成) 和 replicon 免费、无标记的 chromosomal 集成(单个拷贝集成) 被构造。辅酶 Q10 超 Escherichia coli TBW20134 被使用化学上可诱导的 chromosomal 进化， replicon 免费、无标记的 chromosomal 集成以及 menaquinone biosynthetic 小径的删除构造。设计 E。当与 0.075 g 补充了时， coli TBW20134 每辅酶 Q10 的干燥房间质量(DCM ) 的克生产了 10.7 mg ? 瑳摵敩 ? 潣普物 ? 桴 ? 晥敦瑣癩湥獥 ? 景琠敨瀠潲潰敳 ? 潭敤 ? 愠瑬牥慮整椠敤瑮晩捩瑡潩 ? 污潧楲桴 ? 愠摮甠摰瑡 ? 敭桴摯

英文摘要：

The plasmid-expression system is routinely plagued by potential plasmid instability. Chromosomal in-tegration is one powerful approach to overcome the problem. Herein we report a plasmid-free hyper-producer E. coli strain for coenzyme Q10 production. A series of integration expression vectors, pxKC3T5b and pxKT5b, were constructed for chemically inducible chromosomal evolution （multiple copy integration） and replicon-free and markerless chromosomal integration （single copy integration）, respectively. A coenzyme Q10 hyper-producer Es-cherichia coli TBW20134 was constructed by applying chemically inducible chromosomal evolution, replicon-free and markerless chromosomal integration as well as deletion of menaquinone biosynthetic pathway. The engineered E. coli TBW20134 produced 10.7 mg per gram of dry cell mass （DCM） of coenzyme Q10 when supplemented with 0.075 g·L^-1 of 4-hydroxy benzoic acid;this yield is unprecedented in E. coli and close to that of the commercial producer Agrobacterium tumefaciens. With this strain, the coenzyme Q10 production capacity was very stable after 30 sequential transfers and no antibiotics were required during the fermentation process. The strategy presented may be useful as a general approach for construction of stable production strains synthesizing natural products where various copy numbers for different genes are concerned.