利用光合菌Rhodobacter sphaeroides BCRC 13100生產Coenzyme Q10醱酵製程之開發

Process Development of Coenzyme Q10 Production by Rhodobacter sphaeroides BCRC 13100

劉倉豪(Tsang-Hao Liu)

指導教授 ： 莊瑞鑫 ； 魏毓宏

元智大學/工程學院/生物科技暨生物資訊研究所/碩士(2007年)

摘要

本研究目的係利用光合菌Rhodobacter sphaeroides BCRC 13100進行生產輔酶Q10的相關醱酵製程開發，主要著重於培養基及培養系統之建立。
Coenzyme Q10 (輔酶Q10，CoQ10)由於其高抗氧化性可消除體內自由基、活化細胞，達成抗老化之目的，對特定疾病亦有一定的療效，且幾乎無副作用，目前備受重視。
本研究首先利用1H - NMR、FAB-MS及HPLC建立Coenzyme Q10之定性及定量的分析方法。其次由於光合菌具有廣泛且複雜的生理特性，可在各式環境中生長，故本醱酵製程開發分為一般的耗氧醱酵及光合厭氧醱酵進行探討。研究顯示：在光合厭氧醱酵時 ，利用最佳化的培養基對BCRC 13100進行培養，可生產CoQ10濃度與含量分別達29.14 mg/L，6.53 mg/g，相較初期以活化培養基（TSB）進行搖瓶培養，在產量及含量分別有約6倍及2.8倍的提昇。一般耗氧醱酵方面，利用最佳化的生長條件及培養基，有較佳的CoQ10產量，可達31.23 mg/L，菌體內含量則為2.81 mg/g-DCW。
另外，本研究發現，在醱酵同時添加微量成份如生長素Vitamin B1，可提昇CoQ10產量及含量將近100%，達到60.02 mg/L，菌體內含量則為5.25 mg/g-DCW，分別為初始培養的12.6倍及2.26倍。
利用饋料批次的方式進行培養，可得CoQ10產量、菌體乾重及胞內含量分別達103.42 mg/L，18.9 g/L及5.47 mg/g-DCW；再利用重複批次的方式進行培養，可得到菌體乾重37.65 g/L，CoQ10產量138.24 mg/L，含量為3.67 mg/g-DCW。

關鍵字

光合菌 ； Rhodobacter sphaeroides ； 光合厭氧 ； Coenzyme Q10 ； 培養基最適化

並列摘要

Coenzyme Q10 (CoQ10) is a quinone containing ten units of the isoprenoid side chain. Due to it can limit the oxidative attack to the cell, CoQ10 has been used for activating cell and anti-age. It is also useful for specific diseases and almost has no side effect. In recently CoQ10 has been given weight to all bio-products.
The objective of this study was using photosynthetic bacteria, Rhodobacter sphaeroides BCRC 13100, to set up the processes of CoQ10 production. The focuses are to optimize the incubation medium and growth condition.
The CoQ10 was isolated and purified from the broth and was characterized by FAB-MS, 1H-NMR and HPLC. Both 1H-NMR and mass spectrometry results indicate that the purified product was CoQ10.
On the other hand, CoQ10 production in anaerobic fermentation can reach 29.14 mg/L, and content is 6.53 mg/g-DCW (dry cell weight). In aerobic condition, the CoQ10 concentration and dry cell weight is 31.23 mg/L and 2.81 mg/g-DCW, respectively. In this study, after adding some extra factors, like vitamins, amino acids and precursors during fermentation. Only the Vitamin B1 is apparent, which can increase the yield about 100%. The CoQ10 concentration reaches 60.02 mg/L, and content is 5.25 mg/g-DCW.
Using fed-batch fermentation in CoQ10 production, the yield, dry cell weight and content had gained to 103.42 mg/L, 18.9 g/L and 5.47 mg/g-DCW, respectively. Moreover, using repeat fermentation, the yield reached 138.24 mg/L, and the content was 3.67mg/g-DCW.

並列關鍵字

photosynthetic bacteria ； Rhodobacter sphaeroides ； Coenzyme Q10 ； CoQ10 ； ubiquinone ； medium optimization ； fed-batch

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