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標題: 金屬親和吸附材於蛋白質純化及酵素固定化之應用
作者: 何立凡
關鍵字: IMAC;金屬親和吸附;His-tagged protein;protein purification;enzyme immobilization;蛋白質純化;酵素固定化
出版社: 化學工程學系

Several silica-based immobilized metal affinitychromatography
(IMAC) adsorbents with different ligand densities(3-Glymocidoxyprop-
-yltrimethoxysilane-Iminodiacetic acid to Iminodiacetic) were prepared
for the purification of proteins and immobilization of enzymes. The IM-
-AC adsorbents prepared with a ratio of 1:5 exhibit optimal binding cap-
acity for His-tagged proteins and minimal nonspecific adsorption. Results
of SDS-PAGE indicate that the adsorbent can be used for the purification
of His-tagged D-hydatoinase(DHTase) for crude cell lysate in a single
step process with a purify above 90%, comparable to commercial agarose
-based IMAC adsorbents. Due to it's high mechanical strength, the silica-
-based IMAC adsorbents are particularly well suited for industrial
operations. A packed-bed bioreactor with the aforementioned IMAC
adsorbent,loaded with DHTase was used forthe continuous bioconversion
of D,L-p-HPH to N-carbamoyl-D-hydroxyphenylglicine. Under optimal
conditions, 60℃,pH 8,a conversion of 60% was observed at a residue
time of 20 mins. In this study ,we successful demonstrate that a packed-
-bed bioreactor containing silica-based IMAC adsorbents can be
employed for the purification and immobilization of His-tagged enzymes.
Appears in Collections:化學工程學系所

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