Development of an Efficient Enzymatic Process to Synthesize Fructose 1,6-Diphosphate

开发合成果糖 1,6-二磷酸的高效酶法

• 批准号: 06555250
• 负责人: ISHIKAWA Haruo
• 金额: $ 3.46万
• 依托单位: Osaka Prefecture University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06555250/
• 关键词: Hollow-Fiber Reactor; Bucillus stearothermophilus; ATP; Glucokinase; Phosphoglucose Isomerase; Phosphofructokinase; Acetate Kinase; Recycle Number; in-situ方式; アセチルリン酸; グルコース; アセテートコナーゼ

This work was undertaken to develop an efficient enzymatic process to synthesize fructose l, 6-diphosphate (FDP) from glucose and ATP.The principal results were as follows :1.The rates of reactions catalyzed by Bacillus stearothermophilus glucokinase (GK), phosphoglucose isomerase (PGl), phosphofructokinase (PFK) and acetate kinase (AK) were measured under various conditions. By analyzing the data, the mechanisms and the kinetics of the reactions were clarified.2.The enzymatic synthesis of FDP from glucose and the enzymatic ATP regeneration were performed simultaneously in an ultrafiltration hollow-fiber reactor. The process consisted of three-step synthetic reactions catalyzed by GK,PGI and PFK,and the ATP regeneration reaction catalyzed by AK.The experimental results of the yield and the recycle number agreed well with the theoretical predictlons calculated on the basis of the simple analytical model.From the high recycle number and good yields obtained in the present experiment, the enzymatic synthesis of FDP from glucose and the ATP regeneration performed simultaneously in the ultrafiltration hollow-fiber reactor in the diffusion mode was proven to be usable with very good reactor performance.3.The FDP synthesized was purified, and the conditions under which crystalline FDP with a good grade can be obtained were determined.

1.测定了不同条件下嗜热芽孢杆菌葡糖激酶(GK)、磷酸葡萄糖异构酶(PGL)、磷酸果糖激酶(PFK)和醋酸激酶(AK)催化合成果糖L的反应速率。通过对数据的分析，阐明了反应机理和反应动力学。2.在超滤中空纤维反应器中，同时进行了葡萄糖酶促合成FDP和酶促ATP再生的研究。该过程由GK、PGI和PFK催化的三步合成反应和AK催化的ATP再生反应组成。产率和循环次数的实验结果与根据简单分析模型计算的理论预测值吻合较好。从本实验获得的高循环次数和较好的产率来看，在超滤中空纤维反应器中以扩散方式同时进行葡萄糖酶促合成FDP和ATP再生是可行的，具有很好的反应器性能。3.对合成的FDP进行了纯化，确定了获得品级较好的结晶FDP的条件。

项目成果

Haruo Ishikawa, Masahiro Shiroshima, Arief Widjaja, Hiroshi Nakajima and Ryoichi Tsurutani: "Enzymatic Production of Fructose 1,6-Diphosphate and Simultaneous ATP Regeneration in a Hollow-Fiber Reactor" Proceeding of the Regional Symposium on Chemical Eng

Haruo Ishikawa、Masahiro Shiroshima、Arief Widjaja、Hiroshi Nakajima 和 Ryoichi Tsurutani：“在中空纤维反应器中酶法生产果糖 1,6-二磷酸并同时进行 ATP 再生”化学工程区域研讨会论文集

Haruo Ishikawa et al.: "Enzymatic Production of Fructose 1,6-Diphosphate and Simultaneous ATP Regeneration in a Hollow-Fiber Reactor" Proceeding of the Regional Symposium on Chemical Engineering 1996. 381-386 (1966)

Haruo Ishikawa 等人：“在中空纤维反应器中酶促生产果糖 1,6-二磷酸并同时进行 ATP 再生”1996 年化学工程区域研讨会论文集。381-386 (1966)

Haruo Ishikawa et al.: "Production of Fructose 1,6-Diphosphate and Simultaneous ATP Regeneration by Conjugated Enzymes in a Hollow-Fiber Reactor" Proceedings of the 5^<th> World Congress of Chemical Engineering. II. 389-393 (1996)

Haruo Ishikawa 等人：“在中空纤维反应器中通过共轭酶生产果糖 1,6-二磷酸并同时进行 ATP 再生”第五届世界化学工程大会论文集。

Haruo Ishikawa, Masahiro Shiroshima, Arief Widjaja, Hiroshi Nakajima and Ryoichi Tsurutani: "Production of Fructose 1,6-Diphosphate and Simultaneous ATP Regeneration by Conjugated Enzymes in a Hollow-Fiber Reactor" Proceedings of the 5th World Congress of

Haruo Ishikawa、Masahiro Shiroshima、Arief Widjaja、Hiroshi Nakajima 和 Ryoichi Tsurutani：“在中空纤维反应器中通过共轭酶生产果糖 1,6-二磷酸盐并同时进行 ATP 再生”第五届世界大会论文集

Haruo Ishikawa et al.: "Kinetics and Mechanism of Acetate Kinase from Bacillus stearothermophilus" Journal of Chemical Engineering of Japan. 28. 517-524 (1995)

Haruo Ishikawa 等人：“嗜热脂肪芽孢杆菌乙酸激酶的动力学和机制”，日本化学工程杂志。