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

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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.