title: Synergistic effects of chromosomal ispB deletion and dxs overexpression on coenzyme Q(10) production in recombinant Escherichia coli expressing Agrobacterium tumefaciens dps gene.
abstract: 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.
authors.affiliation: Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University, San, Shillim-dong, Kwanak-gu, Republic of Korea.
authors.forename: Jin-Ho
authors.initials: JH
authors.lastname: Choi
authors.forename: Yeon-Woo
authors.initials: YW
authors.lastname: Ryu
authors.forename: Yong-Cheol
authors.initials: YC
authors.lastname: Park
authors.forename: Jin-Ho
authors.initials: JH
authors.lastname: Seo
mesh_terms: D016960:Agrobacterium tumefaciens / Q000201:enzymology* / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D002876:Chromosomes, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D005285:Fermentation / Q000235:genetics; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D000073718:Pancreatitis-Associated Proteins; D014451:Ubiquinone / Q000096:biosynthesis*
chemical_list: D001426:Bacterial Proteins; D000073718:Pancreatitis-Associated Proteins; C000612141:REG3A protein, human; D014451:Ubiquinone