錳(II)及四逆湯對耐輻射奇異球菌於紫外光損傷後之影響

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Title:	錳(II)及四逆湯對耐輻射奇異球菌於紫外光損傷後之影響 The Effect of Mn(II) and Sini-Tang on Deinococcus radiodurans with UV Damage
Authors:	余冠霖
Contributors:	生物科技系鄭淨月
Keywords:	耐輻射奇異球菌四逆湯類胡蘿蔔素電噴灑/大氣壓力化學游離輔助雷射脫附質譜法液相層析質譜儀主成份分析 Deinococcus radiodurans Sini-Tang Carotenoid Electrospray Ionization / Atmospheric Pressure Chemical Ionization / Laser Desorption ionization Liquid Chromatography-Mass Spectrometry Principle Component Analysis
Date:	2013
Issue Date:	2014-03-10 20:41:04 (UTC+8)
Abstract:	之前文獻提出耐輻射奇異球菌（Deinococcus radiodurans﹐Dr）經照射UV後，再添加二價錳離子 Mn(II) 培養時會誘發一些蛋白質表現，使受損的細胞得到修補並有二次生長的現象，其複雜的修復機制十分值得探討。其中類胡蘿蔔素的保護也是一個重要關鍵；這些類胡蘿蔔素正是保護其避免UV、輻射傷害的因素之一。本研究將耐輻射奇異球菌以UV照射後添加四逆湯培養，接著觀察並比較添加 Mn(II) 培養後的差異性。實驗首先利用掃描式電子顯微鏡觀察細胞外受損情形；接著萃取出紅色色素，以薄層層析法(TLC)及液相層析質譜儀(LC/DAD-MS)偵測其表現及變化，並使用自組裝電噴灑/大氣壓力化學游離輔助雷射脫附質譜法(ESI/APCI/LD-MS)進行鑑定；最後再利用基質輔助雷射脫附游離飛行時間質譜儀分析這些紅色色素以及蛋白質的表現差異。結果顯示出 Deinococcus radiodurans﹐Dr 的外觀並無法判斷其受損、恢復的程度，以TLC分析所萃取的紅色色素則有明顯的變化，由LC/DAD-MS發現在400-500nm有微量的吸收，但質譜儀未測得相關訊號。進一步利用ESI/APCI/LD-MS，可檢測出 m/z 567 (M+H)+的訊號，經由MS/MS鑑定後確定這化合物為2-deoxydeinoxanthin。分析Deinococcus radiodurans﹐D r於 UV 損傷後添加 Mn(II) 及四逆湯培養後所萃取之蛋白質，經由主成份分析清楚的顯示出其差異性。 Prior literature suggests that D. radiodurans (Deinococcus radiodurans, Dr) after UV irradiation, and then add the manganese Mn (II) may induce some cultured protein expression, so that repair damaged cells and has been the phenomenon of secondary growth the complexity of the repair mechanism is very worth exploring. Carotenoids which protection is also an important key; these carotenoids protect them from UV and radiation damage of the factors.In this study, D. radiodurans to UV irradiation Add Sini- Tang culture, then observe and compare the added Mn (II) after culture differences. The first experiment was observed by scanning electron microscopy extracellular damage situations; Then extract the red pigment, Thin Layer Chromatography (TLC) and Liquid Chromatography-Mass Spectrometry (LC / DAD-MS) detection of their performance and changes and using Electrospray Ionization / Atmospheric Pressure Chemical Ionization / Laser Desorption ionization (ESI / APCI / LD-MS) were identified; finally re-use Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometer analysis of these red pigments and proteins performance differences.The results show that Deinococcus radiodurans, Dr appearance and can not judge the damage, the degree of recovery to TLC analysis the extracted red pigment is a significant change, the LC / DAD-MS found at 400-500nm with a small amount of absorption, but MS is not related to the measured signal. Further use of ESI / APCI / LD-MS, can be detected m / z 567 (M + H) + signal, via MS / MS to determine which compound was identified as 2-deoxydeinoxanthin. Analysis of Deinococcus radiodurans, Dr to UV damage added after Mn (II) and Sini-Tang cultivation of protein after extraction through principal component analysis clearly shows its difference.
Relation:	電子全文公開日期：20180909，學年度：101，60頁
Appears in Collections:	[Dept. of Biotechnology (including master's program)] Dissertations and Theses

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