Production of coenzyme Q by hydrocarbon-assimilating bacteria.

(pH 7.0) in the following medium: 5% nhexadecane (Tokyo Kasei Co., Tokyo, Japan), 0.1% KH2PO4, 0.4% (NH4)2S04, 0.05% FeSO4 7Hq0, 0.05% MgS04-7H20, and 0.4% yeast extract (Difco). A total of 10.4 g of dry cells was obtained from 2 liters of culture broth in 2 days at 30 C. The CoQ fraction was extracted as described by Lavate et al. (3). A 1-ml amount of the nonsaponifiable extract was taken up in a small volume of benzene for purification by column chromatography on 30 ml of Wako-gel C 200 (Wako Pharmaceutical Co., Osaka, Japan). Benzene was used to elute the distinct yellow fraction containing the CoQ. The CoQ content was assayed by the spectrophotometric method (1). The fraction was then evaporated and developed by thin-layer chromatography with Silica Gel G. The band of CoQ was determined visually under ultraviolet light and eluted with hot ethyl alcohol. Table 1 gives the amounts of CoQ in these fractions. (CoQ6 and CoQio were purchased from Tokyo Kasei Co.) The CoQ was concentrated and crystallized, and the coenzyme was then recrystallized from absolute ethyl alcohol. The recrystalled CoQ showed the same properties as CoQg with respect to the infrared spectrum in KBr, the nuclear magnetic resonance spectrum in CC14, and the melting point of 45 C (5). The hydrocarbon assimilating bacterium, Achromobacter sp., contains CoQg rather than the CoQ that has been found in other microorganisms (2, 4, 6).