文献类型：期刊文献

中文题名：基因替换及多基因共表达强化大肠杆菌辅酶Q_(10)的合成能力

英文题名：Ubiquinone-10 Overproduction in Escherichia coli by Gene Substitution and Multi-gene Coexpression

作者：傅楠[1];叶江[1];吴海珍[1];孔璐[1];张惠展[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

年份：2008

卷号：34

期号：5

起止页码：654

中文期刊名：华东理工大学学报（自然科学版）

外文期刊名：Journal of East China University of Science and Technology

收录：CSTPCD;;EI(收录号：20084811740417);Scopus;北大核心:【北大核心2004】；CSCD:【CSCD2011_2012】；

语种：中文

中文关键词：大肠杆菌;基因缺失;多基因共表达;辅酶Q

外文关键词：Escherichia coli; gene disruption; multigene coexpression; ubiquinone

摘要：为了强化大肠杆菌合成辅酶Q10(CoQ10)的能力,对大肠杆菌进行了相关的基因操作。通过敲除大肠杆菌染色体上的聚八异戊二烯焦磷酸合成酶基因ispB,并导入来自Gluconobactersuboxydans的聚十异戊二烯焦磷酸合成酶基因ddsA,使大肠杆菌具有CoQ10合成能力的同时降低了内源性辅酶Q8(CoQ8)的合成。采用双质粒共表达系统,对CoQ生物合成途径中多个功能基因进行了强化表达,构建得到的重组大肠杆菌CoQ的合成能力、CoQ10合成的专一性都有明显改善,其中ubiCA和ddsA基因的协同表达效果最为明显,CoQ的合成能力比对照提高了65%,而CoQ10的合成量也提高了1.1倍。
In order to overproduce ubiquinone-10, many gene-manipulations were applied to Escherichia coli strain. In this work, the ispB gene of Escherichia coli, which encodes octaprenyl-pyrophosphate synthase was disrupted, while the ddsA gene of Gluconobacter subozydans, which encodes decaprenyl- pyrophosphate synthase was inserted, so that not only ubiquinone-10 was generated, but also the endogenous production of ubiquinone-8 was eliminated in Escherichia coli. Through the dual-plasmid expression systems, several genes were eoexpressed, which played key roles in ubiquinone biosynthesis. Accordingly, the productivity and relative content of ubiquinone-10 were significantly enhanced by the increase of the expression level of ubiCA, and the ddsA genes. The yield of total ubiquinones increased to 1.65 times that of the control, and the actual yield of ubiquinone-10 increased by 110%.