Batch and fed-batch production of coenzyme Q10 in recombinant Escherichia coli containing the decaprenyl diphosphate synthase gene from Gluconobacter suboxydans
Batch and fed-batch production of coenzyme Q10 in recombinant Escherichia coli containing the decaprenyl diphosphate synthase gene from Gluconobacter suboxydans
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DOI：
10.1007/s00253-004-1743-y
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发表时间：
2005-04-01
影响因子：
5
通讯作者：
Seo, JH
中科院分区：
文献类型：
--
作者：
Park, YC;Kim, SJ;Seo, JH
辅酶Q(10) （CoQ(10)）是一种奎宁，由10个类异戊二烯侧链组成。因为它限制了自由基对DNA和脂质的氧化攻击，CoQ（10）已被用作食品、化妆品和药品的抗氧化剂。Decaprenyl diphosphate synthase （DPS）是CoQ（10）中Decaprenyl尾部与异戊烯二磷酸合成的关键酶。在大肠杆菌组成启动子的控制下，表达了编码亚氧化糖葡杆菌DPS的ddsA基因。对重组大肠杆菌BL21/pACDdsA细胞提取物的高效液相色谱和质谱分析表明，CoQ（10）而不是内源性CoQ(8)是主要的辅酶q。低拷贝数的ddsA基因的表达导致CoQ（10）在批量发酵中积累到0.97 mg l（-1）。大肠杆菌BL21/pACDdsA分批补料发酵的高细胞密度（103 g l(-1)）使CoQ（10）浓度达到25.5 mg l(-1)，产率达到0.67 mg l(-1) h(-1)，分别是分批发酵的26.0和6.9倍。
Coenzyme Q(10) (CoQ(10)) is a quinine consisting of ten units of the isoprenoid side-chain. Because it limits the oxidative attack of free radicals to DNA and lipids, CoQ(10) has been used as an antioxidant for foods, cosmetics and pharmaceuticals. Decaprenyl diphosphate synthase (DPS) is the key enzyme for synthesis of the decaprenyl tail in CoQ(10) with isopentenyl diphosphate. The ddsA gene coding for DPS from Gluconobacter suboxydans was expressed under the control of an Escherichia coli constitutive promoter. Analysis of the cell extract in recombinant E. coli BL21/pACDdsA by high performance liquid chromatography and mass spectrometry showed that CoQ(10) rather than endogenous CoQ(8) was biologically synthesized as the major coenzyme Q. Expression of the ddsA gene with low copy number led to the accumulation of CoQ(10) to 0.97 mg l(-1) in batch fermentation. A high cell density (103 g l(-1)) in fed-batch fermentation of E. coli BL21/pACDdsA increased the CoQ(10) concentration to 25.5 mg l(-1) and its productivity to 0.67 mg l(-1) h(-1), which were 26.0 and 6.9 times higher than the corresponding values for batch fermentation.