Batch and fed-batch production of coenzyme Q10 in recombinant Escherichia coli containing the decaprenyl diphosphate synthase gene from Gluconobacter suboxydans.
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.
School of Agricultural Biotechnology, Center for Agricultural Biomaterials, Seoul National University, Seoul, 151-742, South Korea. jhseo94@snu.ac.kr
Yong-Cheol
YC
Park
Soo-Jung
SJ
Kim
Jin-Ho
JH
Choi
Won-Heong
WH
Lee
Kyung-Moon
KM
Park
Mokoto
M
Kawamukai
Yeon-Woo
YW
Ryu
Jin-Ho
JH
Seo
D019883:Alkyl and Aryl Transferases / Q000235:genetics*; D003067:Coenzymes; D004926:Escherichia coli / Q000235:genetics; D005285:Fermentation; D020576:Gluconobacter / Q000201:enzymology*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014451:Ubiquinone / Q000031:analogs & derivatives* / Q000096:biosynthesis*
D003067:Coenzymes; D011994:Recombinant Proteins; D014451:Ubiquinone; D019883:Alkyl and Aryl Transferases; C039821:decaprenyl pyrophosphate synthetase; C024989:coenzyme Q10