For biotechnological production of coenzyme Q(10) (CoQ(10)) in recombinant Escherichia coli, three genetic manipulations were performed: heterologous expression of decaprenyl diphosphate synthase (Dps) from Agrobacterium tumefaciens, deletion of endogenous octaprenyl diphosphate synthase (IspB), and overexpression of 1-deoxy-d-xylulose synthase (Dxs). Expression of the dps gene and deletion of the ispB gene in E. coli BL21(DE3)DeltaispB/pAP1 allowed production of CoQ(10) only. Furthermore, coexpression of the dxs gene increased the specific content of CoQ(10) from 0.55-0.89mgg(-1) to 1.40mgg(-1). For mass production of CoQ(10), fed-batch fermentation of E. coli BL21(DE3)DeltaispB/pAP1+pDXS was carried out in a defined medium with 20gl(-1) initial glucose and by the glucose-feeding strategy of pH-stat. Finally, 99.4mgl(-1) CoQ(10) concentration, 1.41mgg(-1) specific CoQ(10) content and 3.11mgl(-1)h(-1) productivity were obtained in 33h of the fermentation, which were 78, 1.9, and 19 times higher than those for E. coli BL21(DE3)/pAP1 without the ispB deletion and dxs overexpression.
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http://dx.doi.org/10.1016/j.jbiotec.2009.04.010 | DOI Listing |
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