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

超滤中空纤维反应器中共轭酶 G6P 生产和连续 ATP 回收的理论与实验研究。

• 批准号: 60550680
• 负责人: ISHIKAWA Haruo
• 金额: $ 1.41万
• 依托单位: University of Osaka Prefecture
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1985
• 资助国家: 日本
• 起止时间: 1985 至 1986
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-60550680/
• 关键词: ATP; Acetyl Phosphate; Conjugated Enzymes; Bioreactor for Synthesis; Glucokinase; Acetatekinase; Glucose-6-phosphate; ホローファイバーリアクター

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.

为了开发需要ATP作为辅因子的用于酶促合成的有效生物反应器系统，在超滤中空纤维反应器中同时进行以下两个反应：一种是以葡萄糖激酶（GK）为催化剂，由葡萄糖和ATP生成葡萄糖-6-磷酸（G6 P）的主反应，另一种是以乙酰激酶（AK）为催化剂，从乙酰磷酸和ADP再生ATP的反应。两种酶都被包埋在反应器的壳层中，结合底物和少量的辅因子ATP被连续地送入反应管中。数值求解了带反应的传质基本方程，并从葡萄糖转化率和循环数两个方面分析了反应器的性能。在这项工作中，动力学参数和组成成分的渗透率，这是计算所需的，在实验上确定。因此，在适当的条件下，可以预期超过90%的高转化率和超过1000的高循环数。采用模型反应器在不同条件下进行了G6 P的合成。G6 P的合成效率较高，最佳转化率为81%，循环数为1620。这些结果与理论预测相吻合. GK和AK均通过多种方法固定化。其中，通过吸附在离子交换树脂颗粒，伊拉-904，酶的活性，机械强度，价格等点，在未来，与上述固定化酶的填充床反应器的反应器性能进行了实验研究，并与中空纤维反应器，以确定最佳的反应器配置。

项目成果

Haruo Ishikawa: J.Chem.Eng.Journal.

石川春夫：J.Chem.Eng.Journal。

Haruo Ishikawa: "Theoretical Analysis of G6P Production and Continuous ATP Recycling by Conjugated Enzymes in an Ultrafiltration Hollow Fiber Reactor." J. Chem. Eng. Japan.

Haruo Ishikawa：“超滤中空纤维反应器中共轭酶 G6P 生产和连续 ATP 回收的理论分析”。

Haruo Ishikawa: Bull.Soc.Chem.Japan.

石川春夫：Bull.Soc.Chem.Japan。

Haruo Ishikawa: "Experimental Investigation of G6P Production and Continuous ATP Recycling by Conjugated Enzymes in an Ultrafiltration Hollow Fiber Reactor." J. Chem. Eng. Japan.

Haruo Ishikawa：“超滤中空纤维反应器中共轭酶生产 G6P 和连续 ATP 回收的实验研究”。

Haruo Ishikawa: "Initial Rate Studies of a Thenmophilic Glucokinase from Bacillus Stearothermophilus." Bull. Soc. Chem. Japan.

Haruo Ishikawa：“来自嗜热脂肪芽孢杆菌的嗜热葡萄糖激酶的初始速率研究。”