Mannens G, Slegers G
Department of Pharmaceutical Sciences, State University of Ghent, Belgium.
Biochim Biophys Acta. 1990 Mar 29;1038(1):68-73. doi: 10.1016/0167-4838(90)90011-4.
The enzyme arylamine acetyltransferase (acetyl-CoA:arylamine N-acetyltransferase, EC 2.3.1.5) from pigeon liver is immobilized onto differently derivatized controlled pore glass beads. Different silanes, spacer arms and reactive end-groups were tested, and immobilized enzyme stability tests were performed. From these experiments, the method of choice was selected: immobilization on controlled pore glass beads (24 nm pore size, 75-125 microns particle size) derivatized with gamma-aminopropyl and glutaraldehyde as the reactive end group. The kinetic properties of an enzyme reactor were investigated and optimized. The goal was to obtain a rapid high-yield conversion of 0.5-1 mumol acetyl-CoA to N-acetylserotonin, so that the reactor is useful for the 11C-labelling of N-acetylserotonin. Using an enzyme reactor (9.8 x 0.5 cm i.d.) containing 4.6 U active arylamine acetyltransferase immobilized onto 930 mg carrier, a 70% conversion of acetyl-CoA was obtained within 4 min.
将来自鸽肝的芳胺乙酰基转移酶（乙酰辅酶A：芳胺N - 乙酰基转移酶，EC 2.3.1.5）固定在不同衍生化的可控孔径玻璃珠上。测试了不同的硅烷、间隔臂和反应性端基，并进行了固定化酶的稳定性测试。通过这些实验，选择了以下方法：固定在孔径为24 nm、粒径为75 - 125微米的可控孔径玻璃珠上，用γ-氨丙基进行衍生化，并以戊二醛作为反应性端基。对酶反应器的动力学特性进行了研究和优化。目标是实现0.5 - 1微摩尔乙酰辅酶A快速高产率地转化为N - 乙酰血清素，从而使该反应器可用于N - 乙酰血清素的11C标记。使用一个内径为9.8×0.5 cm的酶反应器，其中930 mg载体上固定有4.6 U活性芳胺乙酰基转移酶，在4分钟内乙酰辅酶A的转化率达到了70%。
