耐輻射奇異球菌 ( Deinococcus radiodurans, Dr ) 經由紫外光照射後，再添加二價錳離子Mn2+培養時會使受損的細胞得到修補並有二次生長的現象。除了特定蛋白質表現外，類胡蘿蔔素等二次代謝物也會隨著二次生長而有不同的表現，這些化合物是保護其避免UV、輻射傷害的因素之一。 因此本研究將耐輻射奇異球菌以UV照射後添加二價錳離子Mn2+培養，實驗首先由肉眼觀察出菌液的顏色變化，接著利用乙酸乙酯萃取培養液的二次代謝物，並收集菌體使用丙酮萃取類胡蘿蔔素之後，利用薄層層析法觀察類胡蘿蔔素的變化，再利用高效能液相質譜法(HPLC/MS)分析其二次代謝物及類胡蘿蔔素，來偵測其化合物表現與變化，並以統計方法尋找代謝生物標記。實驗結果發現當添加Mn2+培養時，有多種的二次代謝物和類胡蘿蔔素表現量增加。利用HPLC/MS/MS可檢測出類胡蘿蔔素訊號m/z 566.5 (M+H)+、Deinoxanthin ( m/z 583.5 (M+H)+ )、3',4'-dihydrodeinoxanthin ( m/z 585.5 (M+H)+ )有明顯的增加，其中m/z 566.5 (M+H)+的含量為最多，經由高解析質譜確認這化合物為2-deoxydeinoxanthin ( m/z 566.4124 (M+H)+ )。而半定量結果顯示Dr Mn類胡蘿蔔素濃度有明顯的增加。 二次代謝物經由HPLC/MS/MS可檢測出訊號 m/z 100.1 (M+H)+ 、m/z 123.0 (M+H)+、m/z 169.0 (M+H)+、m/z 197.0 (M+H)+、m/z 211.1 (M+H)+、m/z 245.0 (M+H)+有明顯的增加，而m/z 227.0 (M+H)+、m/z 557.0 (M+H)+為Dr生長時表現出來的訊號，而m/z 220.0 (M+H)+、m/z 295.0 (M+H)+、m/z 328.0 (M+H)+為Dr 加Mn2+培養時質譜偵測與Dr比較所特定表現出來的訊號。以半定量結果顯示Dr Mn二次代謝物濃度有明顯的增加。 Deinococcus radiodurans after UV irradiation, add the manganese Mn (II) may repair damaged cells and has been the phenomenon of secondary growth. In addition to expression of specific proteins, the carotenoids and other secondary metabolites with secondary growth will have different expression, these compound protect them from UV and radiation damage of the factors.Here we show that the carotenoids contributes to the resistance of D. radiodurans to UV irradiation in manganese Mn (II) culture. Carotenoids of the bacterium were extracted with acetone, and the secondary metabolites of the culture were extracted with ethyl acetate. The performance of carotenoids and secondary metabolites in Mn-accumulating bacteria were analyzed by Thin Layer Chromatography (TLC) and High Performance Liquid Chromatography-Mass Spectrophotometer (HPLC-MS). During recovery from UV irradiation, the levels of carotenoids increase, consistent with a role in repairing radiation damage. The predominant peak of m/z 566.5 (M+H)+ was identified as 2-deoxydeinoxanthin via high resolution mass spectrometer (m/z 566.4124).
