微生物鑑定是分類學的一部份，而快速且有效的微生物鑑定在疾病診斷、病原菌所釋放的有害物質之鑑定、食品污染監管及生物製程調控上的應用是相當重要且必須的。近年來，由於離子化技術的改良，質譜儀已可提供生物分子，如DNA、蛋白質、醣類、脂質，這類大型分子的分析，加上其具有分析快速、靈敏度佳及準確性高等特性，因此在微生物鑑定的應用上極具潛力。
本研究主要目的為利用基質輔助雷射脫附游離(matrix assisted laser desorption/ionization, MALDI)質譜分析的方式，偵測可作為放線菌分類的標的蛋白質(Biomarker)，並進一步利用質譜技術將標的蛋白質加以定性。本實驗選用六屬標準放線菌株作為分析對象，每株皆以全細胞及細胞萃取液的形式，進行基質輔助雷射脫附游離質譜技術分析。經由質譜分析的結果，我們找到其中二屬的標的蛋白質，分別為Microbispora的m/z 7028及Dactylosporagium 的m/z 7223。利用質譜技術定性蛋白質的方式主要分為胜月太圖譜分析及胜月太序列分析。首先兩個標的蛋白質先以管柱層析的方式從兩屬菌株中逐步純化出，經由蛋白質水解酵素切割後，再利用基質輔助雷射脫附游離質譜儀及液相層析質譜儀進行胜月太圖譜(peptide mapping)分析。為了準確定義標的蛋白質，進一步利用傳統的Edman degradation、MALDI-post source decay(PSD)及Electrospray-collision- induced dissociation(ESI-CID)方式進行月太序列分析。

質譜分析的結果經由資料庫搜尋比對後發現，我們所偵測出的標的蛋白質皆為cold shock protein。本實驗結果顯示質譜分析的方式應可作為一靈敏且準確的菌種鑑定方法，我們期望運用此一技術偵測更多放線菌的cold shock protein，應用於放線菌分類上。

Intact actinomycetes and their cellular extracts were analyzed directly on a matrix-assisted laser desorption/ionization time-of-flight（MALDI-TOF）mass spectrometer. Experiments were designed to identify biomarkers from different genera of actinomycetes. A biomarker of 7026 ± 2 Da was detected in the mass spectra of three species that belong to the genus Microbispora. Another polypeptide of 7222 ± 1 Da was identified in two species of actinomycetes from the genus Dactylosporangium. After chromatography separations, the biomarkers from Microbispora and Dactylosporangium were subjected to protease digestion and ‘peptide mapping’ analysis. The results were used in database search. Furthermore, partial sequencing data were obtained by conventional Edman degradation and analyses of post source decay (PSD) and collision induced dissociation (CID) fragment ion spectra. From these analyses, we suggest that these biomarkers are cold shock proteins and mass spectrometry can provide a sensitive and accurate method for identifying microorganisms. Therefore, with mass spectrometric analysis, we hope to identify more cold shock proteins as biomarkers for classification of actinomycetes.

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