Theoretical adn Experimental Investigation of G6P Production and Continuous ATP Recycling by Conjugated Enzymes in an Ultrafiltration Hollow Fiber Reactor.

为了开发一种高效的以ATP为辅助因子的酶促合成生物反应器系统，在超滤中空纤维反应器中同时进行了以下两个反应：一个是葡萄糖激酶（GK）为催化剂，葡萄糖和ATP生成葡萄糖-6-磷酸（G6P）的主反应，另一个是乙酰磷酸（AK）为催化剂，乙酰磷酸和ADP再生ATP的反应。这两种酶都被包裹在反应器的外壳中，共轭底物和少量的辅因子ATP被连续地送入管中。主要研究结果如下：1。对反应传质的基本方程进行了数值求解，并从葡萄糖转化率和循环次数两方面分析了反应器的性能。本文通过实验确定了计算所需的动力学参数和组成组分的渗透率。因此，在适当的条件下，可以预期90%以上的高转化率和1000以上的高回收次数。利用模拟反应器在不同条件下进行了G6P的合成。G6P的合成效率最高，转化率为81%，回收率为1620%。这些结果与理论预测相吻合。GK和AK均通过几种方法固定。其中，将酶吸附在离子交换树脂颗粒中制备的IRA-904从酶活性、机械强度、价格等方面都是最好的。在未来，将对上述固定化酶的填充床反应器性能进行实验研究，并与中空纤维反应器进行比较，以确定最佳反应器配置。

In order to develop an efficient bioreactor system for enzymatic synthesis which requires ATP as a cofactor, the following two reactions were carried out simultaneously in an ultrafiltration hollow fiber reactor: one is a main reaction that produces glucose-6-phosphate (G6P) from glucose and ATP with glucokinase (GK) as a catalyst, and the other is a reaction that regenerates ATP from acetyl phosphate and ADP with acetatekinase (AK) as a catalyst. Both the enzymes are entrapped in the shell of the reactor, and the conjugated substrates and a small amount of the cofactor ATP are fed continuously into the tubes.The main findings are as follows:1. Fundamental equations for mass transfer with reactions were solved numerically, and the performance of the reactor was analyzed in terms of the conversion of glucose and the recycle number. The kinetic parameters and the permeabilities of the constituting components, which were required for calculation, were determined experimentally in this work. As a result, one can expect a high conversion of more than 90 % and high recycle number of more than 1000 under appropriate conditions.2. The synthesis of G6P was carried out under various conditions by using a model reactor. The G6P was synthesized efficiently, the best results being 81 % for conversion and 1620 for recycle number. These results agreed well with the theoretical predictions.3. Both the GK and AK were immobilized by several methods. Among them, the one made by adsorbing the enzymes in the ion exchange resin particles, IRA-904, was the best from the point of the enzyme activity, mechanical strength, price, etc. In the future, the reactor performance of a packed bed reactor with the above immobilized enzymes will be examined experimentally and be compared with that of the hollow fiber reactor in order to determine the best reactor configuration.