Study On Extraction Technology Of High Purity Of Solanesol From Waste Tobacco And CoQ₁₀ Fermentation

CoQ₁₀,which is also called ubiquinone,is an important transferringhydrogen carrier of biological respiratory chain.So it has a significant influence on human beings.To date,it has been widely used in the treatment of cardio-cerebrovascular disease,hypertension,nervous system and skin diseases.It is also used in the preparation of cosmetic products and health products.Solanesol is an important side chain precursor of CoQ₁₀.In this paper,the purifying technology of high purity of solanesol, the fermentation technology of the CoQ₁₀ and fermentation promotion material of CoQ₁₀ was studied.Firstly,a novel process to extract and purify solanesol form discard tobacco leaves was introduced,which contains five steps as follows: extraction,dewaxing,saponification,crystallization and recrystallization, without column chromatographic step in traditional process.The purity of final solanesol reached 94.35%.Then the effect of the glucose and pH change on CoQ₁₀ production by Sphingomonas sp.ZUTEO3 was studied in Flask.The optimal technological conditions was as follows:initial sugar concentration of fermentation as 10 g/L,pH was adjusted to 6 after 20 h fermentation,and 0.10%(w/v) glucose was added after 30 h fed-batch fermentation,the CoQ₁₀ yield could reach 1.67 mg/L after 48 h fermentation.It was scaled up successfully in 5L fermenter,in which the CoQ₁₀ yield could reach 2.07 mg/L after 54 h fermentation.Crude extract of CoQ₁₀ was purificated by silica gel column chromatography following by recrystallizing.The purity of recrystallized sample was 17.63%.So,further purification is necessary in the future research.The effect of the addition of crude solanesol,saponified sample and 99%solanesol on CoQ₁₀ production was studied respectively.The results indicated that the crude solanesol showed the best effect to improve the CoQ₁₀ production and the yield could reach 3.11 mg/L.Then the crude solanesol was separated by silica gel column to obtain compotnent except solanesol,which were added into broth to confirm their effects on the promotion effect on CoQ₁₀ fermentation.It was found that the components in the blots below the solanesol blot on silica gel sheet showed the best effect on CoQ₁₀ fermentation,the CoQ₁₀ yield reached 11.73 mg/L.The promotion-active material was fuether separated and purified by silica gel column and detected by HPLC and GC-MS.The results showed the main components belonged to butylated hydroxytoluene and linolenic acid.