Amplification of 1-deoxy-d-xyluose 5-phosphate (DXP) synthase level increases coenzyme Q10 production in recombinant Escherichia coli
- 257 Downloads
For the enhancement of coenzyme Q10 (CoQ10) production, 1-deoxy-d-xylulose 5-phosphate (DXP) synthase of Pseudomonas aeruginosa was constitutively coexpressed in a recombinant Escherichia coli strain, which harbors the ddsA gene from Gluconobacter suboxydans encoding decaprenyl diphosphate synthase. It was found that the expression of the ddsA gene caused depletion of the isopentenyl diphosphate (IPP) pool in E. coli. Amplification of DXP synthase level by installing P. aeruginosa DXP synthase restored the diminished IPP pool and concomitantly resulted in approximately a twofold increase in relative content and productivity of CoQ10. Maximum CoQ10 concentration of 46.1 mg l−1 was achieved from glucose-limited fed-batch cultivation of the recombinant E. coli strain simultaneously harboring the ddsA and dxs genes.
KeywordsLycopene CoQ10 Isopentenyl Diphosphate CoQ10 Production CoQ10 Content
This work was supported by Korea Ministry of Commerce, Industry and Trade, and Ministry of Education through the BK21 program and in part by Korea Research Foundation Grant funded by Korean government (KRF-2004-005-F00055).
- Kawamukai M (2002) Biosynthesis, bioproduction and novel roles of ubiquinone. J Biosci Bioeng 94:511–517Google Scholar
- Kuzuyama T, Takagi M, Takahashi S, Seto H (2000) Cloning and characterization of 1-deoxy-d-xylulose 5-phosphate synthase from Streptomyces sp. strain CL190, which uses both the mevalonate and nonmevalonate pathways for isopentenyl diphosphate biosynthesis. J Bacteriol 182:891–897CrossRefGoogle Scholar
- Lois LM, Campos N, Rosa Putra S, Danielsen K, Rohmer M, Boronat A (1998) Cloning and characterization of a gene from Escherichia coli encoding a transketolase-like enzyme that catalyzes the synthesis of d-1-deoxyxylulose 5-phosphate, a common precursor for isoprenoid, thiamin, and pyridoxal biosynthesis. Proc Natl Acad Sci U S A 95:2105–2110CrossRefGoogle Scholar
- Negishi E, Liou SY, Xu C, Huo S (2002) A novel, highly selective, and general methodology for the synthesis of 1,5-diene-containing oligoisoprenoids of all possible geometrical combinations exemplified by an iterative and convergent synthesis of coenzyme Q10. Org Lett 4:261–264CrossRefGoogle Scholar
- Nichols BP, Green JM (1992) Cloning and sequencing of Escherichia coli ubiC and purification of chorismate lyase. J Bacteriol 174:5309–5316Google Scholar
- Szkopiñska A (2000) Ubiquinone. Biosynthesis of quinone ring and its isoprenoid side chain. Intracellular localization. Acta Biochim Pol 47:469–480Google Scholar