| 大肠杆菌链长控制基因ddsA原位替换及多基因共表达对辅酶Q_(10)合成能力的影响 |
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| 引用本文： | 孔璐,傅楠,叶江,吴海珍,张惠展. 大肠杆菌链长控制基因ddsA原位替换及多基因共表达对辅酶Q_(10)合成能力的影响[J]. 食品与药品, 2009, 0(9) |
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| 作者姓名： | 孔璐 傅楠 叶江 吴海珍 张惠展 |
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| 作者单位： | 华东理工大学生物反应器工程国家重点实验室; |
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| 摘 要： | 目的强化大肠杆菌合成辅酶Q10(CoQ10)的能力。方法利用途径工程的基本原理,对大肠杆菌辅酶Q生物合成途径中链长控制基因ispB进行遗传操作,并强化表达该途径中多个功能基因。结果首次成功用来自Gluconobacter suboxydans的十聚异戊二烯焦磷酸合成酶基因ddsA原位替换大肠杆菌染色体上ispB基因,构建得到原位替换株JKL;同时强化表达CoQ生物合成途径的相关基因,发现在ubiCA和ddsA协同表达的重组菌pDCA/JKL中CoQ的合成能力比JKL提高了2.3倍,CoQ10合成量提高了2.5倍。检测该重组菌中ubiA的转录水平,发现其mRNA相对拷贝数是对照JKL的125倍。结论成功获得的重组大肠杆菌在降低内源性CoQ8合成能力的同时具备合成较长侧链CoQ10的能力,且通过强化表达相关基因使合成CoQ10的能力得到了提高。
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| 关 键 词： | 大肠杆菌 ddsA 基因替换 共表达 辅酶Q10 qRT-PCR |
Influence of Gene Substitution with Gluconobacter Suboxydans ddsA and Multi-gene Co-expression on Production of Ubiquinone-10 in Escherichia coli |
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| KONG Lu,FU Nan,YE Jiang,WU Hai-zhen,ZHANG Hui-zhan. Influence of Gene Substitution with Gluconobacter Suboxydans ddsA and Multi-gene Co-expression on Production of Ubiquinone-10 in Escherichia coli[J]. Food and Drug, 2009, 0(9) |
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| Authors: | KONG Lu FU Nan YE Jiang WU Hai-zhen ZHANG Hui-zhan |
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| Affiliation: | State Key Lab of Bioreactor Engineering;East China University of Science and Technology;Shanghai 200237;China |
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| Abstract: | Objective To overproduce ubiquinone-10 in Escherichia coli. Methods By the principle of pathway engineering, a series of gene manipulations of ispB gene which encodes octaprenyl pyrophosphate synthase in the ubiquinone biosynthesis of E. coli JM83 were taken. Also several genes were overexpressed, which played a central role in this metabolic pathway. Results The ispB gene of Escherichia coli JM83, which encodes octaprenyl pyrophosphate synthase in the ubiquinone biosynthesis, was first successfully replace... |
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| Keywords: | Escherichia coli ddsA gene substitution co-expression ubiquinone-10 qRT-PCR |
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