橙黃壺菌BL10已知能透過一種蛋白的分泌來調控細胞群聚的發展。將疑似外泌蛋白數個上游基因利用E. coli表現，進行活性分析發現只有#3413重組蛋白具與外泌蛋白一致的活性。然而該重組蛋白的水溶性不佳，推測是因外泌蛋白具醣基修飾；來自E. coli表達的重組蛋白可能因缺乏醣基而導致水溶性不佳。本研究的目的，在確認外泌蛋白是否為醣蛋白，及S. pombe表現系統是否能用以製造出水溶性較佳的#3413重組蛋白。
為確認外泌蛋白是否具醣基修飾，將agar plate萃取所得含外泌蛋白的粗萃物以離子交換樹脂移除agar的多醣後，利用SDS-PAGE、醣蛋白染劑分析，確實在經Schiff's reagent染色後看到明顯呈色的外泌蛋白訊號，說明其為醣蛋白的機會很高。
利用S. pombe表達#3413重組蛋白，經各式培養及誘導條件的優化，已可誘導足夠#3413重組蛋白，然其水溶性仍不佳，僅能利用高濃度尿素、提高緩衝液pH來提高溶解度，且此狀態的蛋白無法吸附於nickel column，以致無法套用on-column refolding完成重組蛋白的摺疊與純化，說明醣基化非影響重組蛋白水溶性的關鍵；在培養基內發現水溶性佳，然分子量低於預期的活性蛋白或許經過轉譯後修飾、移除部分片段形成的一種蛋白。後續將嘗試透過一些序列比對及結構預測的方法探究其可能性。

Aurantiochytrium sp. strain BL10 is known to regulate the development of cell clustering through the secretion of a protein. Using E. coli express several upstream genes of secretion protein, the activity analysis showed that only the #3413 recombinant protein had the same activity as the secretion protein. However, the poor water solubility of the recombinant protein is presumably due to the glycosylation modification; the recombinant protein expressed from E. coli may be poorly water soluble due to lack of glycosylation. The purpose of this study was to determine whether the secreted protein is a glycoprotein, and whether the S. pombe expression system can be used to produce the #3413 recombinant protein with better water solubility. In order to confirm whether the secretion protein has glycosyl modification, the crude extract containing the secreted protein extracted from the agar plate is analyzed by SDS-PAGE and glycoprotein stain. The secreted protein showed a clear color reaction after staining with Schiff's reagent, indicating that it has a high chance of being a glycoprotein. Using S. pombe to express #3413 recombinant protein, after optimization of various culture and induction conditions, enough #3413 recombinant protein can be induced, but its water solubility is still poor. Its can only be used add high concentration of urea and increase the pH of the buffer solution to improve solubility. The protein in this state cannot be adsorbed on the nickel column, so that on-column refolding cannot be applied. Glycosylation is not the key to affecting the water solubility of the recombinant protein; An active protein with a molecular weight lower than expected may be a protein formed by post-translational modification and removal of partial fragments. In the future, we will try to explore its possibilities through some sequence alignment and structure prediction methods.

沈筱凌，橙黃壺菌BL10之多元不飽和脂肪酸合成酶的純化，國立成功大學生物科技研究所碩士論文，2016。

周思丞，探討橙黃壺菌BL10品系阿米巴細胞形成與遷移的機制，國立成功大學生物科技研究所碩士論文，2013。

陳宛渝，橙黃壺菌 BL10 品系之外泌蛋白純化與功能分析，國立成功大學生物科技研究所碩士論文，2015。

劉致寧，橙黃壺菌 L-BL10 品系之聚酮合成酶基因序列與表現分析，國立成功大學生物科技研究所碩士論文，2013。

謝采妮，橙黃壺菌 BL10 遷移及分化因子的研究，國立成功大學生物科技與產業科學系碩士論文，2018。

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