醣類吸附模組 (carbohydrate-binding modules, CBMs) 為一種可與碳水化合物進行特定的辨識與結合之蛋白區段。米根黴菌 (Rhizopus oryzae) 葡萄糖澱粉酵素 (glucoamylase) 之胺基端具有一澱粉吸附區 (starch-binding domain, RoSBD)，隸屬於CBM家族二十一，且對天然澱粉與可溶性多醣類具有高度結合力。RoSBD之立體結構已利用核磁共振與晶體繞射法解析，有八個β-strands並形成兩個β-sheets以及一個傾斜的木桶狀構造。此結構中含有兩個配體結合位：第一個結合位由色胺酸47、酪胺酸83、酪胺酸93、及酪胺酸94組成，第二個結合位則由酪胺酸32、苯丙胺酸58、及酪胺酸67組成。
CBMs可以單一區域或串聯的形式存在於不同的蛋白當中。一般而言，串聯的CBM較單一之CBM的醣類結合能力高。本實驗室已成功利用大腸桿菌 (E. coli) 之表現系統表現雙聚體RoSBD。為了區分在雙聚體RoSBD中四個關鍵配體結合位之重要性，將其中主要的與配體結合之胺基酸酪胺酸32與色胺酸47分別突變為丙胺酸。本研究發現將雙聚體RoSBD的兩個色胺酸47同時突變後，其對於不可溶澱粉之結合能力明顯地下降。另一方面，利用恆溫滴定熱量計(isothermal titration calorimetry, ITC) 測量雙聚體RoSBD與不同可溶醣類的結合力，其結果顯示將雙聚體RoSBD的兩個酪胺酸32同時突變後會喪失對於可溶醣類的結合能力。此外，雙聚體RoSBD之分子結構可利用另一個已有結構之RoCBM21 (CP90) 為模板預測其結構與功能之相關性。了解雙聚體RoSBD和醣類的結合模式讓我們能更深入地探討串聯之CBM與不同醣類結合之分子機制。
　　由於RoSBD在pH 5~8的環境中對於直鏈澱粉的結合能力較高，在pH 10~11的環境則對直鏈澱粉的結合能力明顯地下降，因此具有RoSBD的目標重組蛋白可利用不同pH值的緩衝溶液在直鏈澱粉純化樹脂中純化回收。α-1酸性醣蛋白 (alpha-1-acid glycoprotein, AGP) 是一種急性期反應蛋白，主要在人體中的肝臟細胞中合成。其pI值約在2.8~3.8之間，是高度醣基化的蛋白，醣類約佔其總重量的45%。AGP具有三項主要功能，包括抗發炎、免疫調節以及與藥物結合，其與藥物結合時對對掌性藥物具專一性篩選能力。
　　本研究成功地利用酵母菌 (Pichia pastoris KM71) 表現重組蛋白RoSBD-AGP，再利用蛋白質水解酶將重組蛋白RoSBD-AGP中的RoSBD移除並分離，進而得到重組的單一AGP。利用色胺酸螢光遞減測試 (Tryptophan fluorescence quenching assay) 確認由RoSBD純化之重組AGP具有與藥物結合之能力，未來可繼續開發對掌性藥物篩選及分離之新穎材料。

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