Production Of Coenzyme Q₁₀ By Fermentation With Blakeslea Trispora And Its Metabolized Regulation

Biosynthetic pathway of coenzyme Q₁₀ in Blakeslea trispora and its metabolized regulation were primarily studied in this study. The strain of Blakeslea trispora JSF4 was treated with ultraviolet and ⁶⁰Co irradiation. By re-screening, the mutant JSF4-JB1, with dual resistant markers, was obtained on the L-Methionine, D-tyrosine and p-aminophenylalanine resistant culture media. Its coenzyme Q₁₀ yield was improved 92% as the parent strain, reaching 27.1 mg/L.The fermentation conditions of Blakeslea trispora JSF4- JB1 was studied, and the optimum culture medium was obtained as follows(g/L): glucose 19, pease 30, peptone 15, KH2PO4 1, MgSO4 0.1, MnSO4 0.4, vitamin B1 0.01, bean oil 9, cottonseed oil 18 and the originally pH 6.5. The mutant was cultured on the rotating shaking bed at the speed of 200 r/min under 25℃for 72 hours,with inoculum size 8% and medium volume 40 mL/500 mL. The results showed that the coenzyme Q₁₀ could be improved 35%, reaching 36.69 mg/L.The extracting coenzyme Q₁₀ from Blakeslea trispora using saponific ation was also studied by orthogonal test. And the optimum extracting conditions were obtained as follows: Wet culture was distilled at 70℃for 60min, added caustic potash up to 1.9 times of dry culture(w/w), pyrogallol up to 1/2 dry culture(w/w), and the concentration of alkali-methanol was 10%.The average recovery of CoQ₁₀ was 81.5%.Coenzyme Q₁₀ was separated by Thin-layer Chromatograph, with its average recovery of 82.8%, and was determined by Ultraviolet Spectrameter and High Performance Liquid Chromatograph. The results showed it a fast and accurate determination for CoQ₁₀.