Appl Microbiol Biotechnol (2005) 67: 192–196 DOI 10.1007/s00253-004-1743-y BIOTECHNOLOGICAL PRODUCTS AND PROCESS ENGINEERING Yong-Cheol Park . Soo-Jung Kim . Jin-Ho Choi . Won-Heong Lee . Kyung-Moon Park . Mokoto Kawamukai . Yeon-Woo Ryu . Jin-Ho Seo Batch and fed-batch production of coenzyme Q 10 in recombinant Escherichia coli containing the decaprenyl diphosphate synthase gene from Gluconobacter suboxydans Received: 6 July 2004 / Revised: 26 August 2004 / Accepted: 31 August 2004 / Published online: 1 October 2004 # Springer-Verlag 2004 Abstract 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, cosmet- ics 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 ex- pressed under the control of an Escherichia coli constitu- tive promoter. Analysis of the cell extract in recombinant E. coli BL21/pACDdsA by high performance liquid chro- matography and mass spectrometry showed that CoQ 10 rather than endogenous CoQ 8 was biologically synthe- sized 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. Introduction Coenzyme Q 10 (CoQ 10 ; 2,3-dimethyl-5-methyl-6-decapre- nyl-1,4-benzoquinone) is an oil-soluble quinone contain- ing ten units of the isoprenoid side-chain. CoQ 10 is used as an antioxidant for cosmetics and pharmaceuticals, because of its role in protecting membrane phospholipids, lipopro- teins and DNA from free radical-induced oxidative damage (Szkopińska 2000). Furthermore, it prevents cardiovas- cular disease and mitochondrial respiratory-chain diseases (Sarter 2002; Geromel et al. 2002). CoQ 10 is embedded in the mitochondrial inner membranes of all human tissues (Szkopińska 2000) and Schizosaccharomyces pombe (Saiki et al. 2003), and in the cellular membranes of Glucono- bacter suboxydans (Okada et al. 1998), Paracoccus de- nitrificans (Takahashi et al. 2003) and Agrobacterium tumefaciens (Lee et al. 2004). Decaprenyl diphosphate synthase (DPS) catalyzes the condensation reaction of isopentenyl diphosphate with al- lylic diphosphate and is a key enzyme to synthesize the decaprenyl tail in CoQ 10 . A gene encoding DPS was iden- tified recently in several microbes, such as S. pombe, G. suboxydans, Aspergillus clavatus, Leucosporidium scotti, and Agr. tumefaciens (Suzuki et al. 1997; Okada et al. 1998; Matsuda et al. 2003; Lee et al. 2004). Most DPS enzymes had a homology of 30–50% to other polyprenyl diphos- phate synthases. The ddsA gene from G. suboxydans con- sisted of 948 bp and encoded a 33,898-Da DPS (Okada et al. 1998). Recombinant expression of the ddsA gene compen- sated the deficiency of coenzyme Q 8 (CoQ 8 ) production by loss of the octaprenyl diphosphate synthase gene (ispB) in Escherichia coli. The selection of microorganisms producing CoQ 10 and gene cloning for DPS and its characterization have been carried out (Suzuki et al. 1997; Okada et al. 1998; Lee et al. 2004). But process development reported that chemi- Y.-C. Park . S.-J. Kim . J.-H. Choi . W.-H. Lee . J.-H. Seo (*) School of Agricultural Biotechnology, Center for Agricultural Biomaterials, Seoul National University, Seoul, 151-742, South Korea e-mail: jhseo94@snu.ac.kr Tel.: +82-2-8804855 Fax: +82-2-8735095 K.-M. Park Department of Chemical System Engineering, College of Science and Technology, Hongik University, Jochiwon, Chungnam, 339-701, South Korea Y.-W. Ryu Department of Molecular Science and Technology, Ajou University, Suwon, 442-749, South Korea M. Kawamukai Department of Life Science and Biotechnology, Faculty of Life & Environmental Science, Shimane University, Matsue 690-8504, Japan