醣祿錠是一種阿爾發－葡萄糖甘酶的抑制劑，已被廣泛用來治療糖尿病患，ATase酵素（acarviosyl transferase）能轉換醣祿錠衍生物上的acarviosyl官能基，可以提高醣祿錠產率及純度，是量產醣祿錠所需之重要酵素。為了更方便地取得ATase酵素，我們從Actinoplanes sp.菌種中選殖出ATase酵素基因，並將其接進帶有SpA訊息序列和ZZ domain的載體，並在大腸桿菌表現含有protein A片段ZZ domain和ATase酵素之融合蛋白（ZZ-ATase），然後以IgG 管柱純化此融合蛋白。本研究更進一步利用薄層層析法（TLC，Thin Layer Chromatography）及高效能液相層析儀( HPLC，High Performance Liquid Chromatography )測定ZZ-ATase酵素之活性、最佳反應條件並測出其酵素動力學常數（和acarbose的Km為0.123 mM，Vmax為0.469 mM/hr）。另外，也改變實驗醣類，測試重組後的ATase酵素是否會廣泛地選擇受質而產生新的產物，結果顯示重組後的ATase酵素僅對某些醣類具有轉換acarviosyl官能基的活性。

Acarbose is an inhibitor of α-glucosidase and has been widely used to treat patients with diabetes. Acarbose is a metabolite of Actinoplanes sp. and it can be obtained by microbial fermentation method. Acaviosyl transferase (ATase) is an enzyme that can hydrolyze and transfer the acarviosyl group of acarbose. It has been proposed that ATase may increase the yield and purity of acarbose. In order to evaluate the potential of using recombinant ATase for acarbose production, we have cloned the ATase gene from Actinoplanes sp. and expressed a fusion protein containing the N-terminus of protein A (ZZ domain) and the full length of ATase in E. coli. The fusion protein ZZ-ATase was purified to homogeneity by IgG affinity chromatography. The ATase activity of the fusion protein has been evaluated by thin layer chromatography and comfirmed by HPLC method more precisely. The parameters of enzyme kinetics measured by HPLC showd that the Km= 0.123 mM and Vmax is 0.469 mM/hr. We changed other substrates to see whether the ZZ-ATase has broad-acceptor activity. The result revealed that ZZ-ATase can only transfer acarviosyl unit to a few substrates such as salicin and cellobiose.

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