1993 Fiscal Year Final Research Report Summary
Stabilization and Activation of Enzymes by Adsorptive Immobilization with Water-Soluble Polymers and Its Application to Enzyme Membrane Reactor
| Project/Area Number |
04555229
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
反応工学
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| Research Institution | Kyoto Institute of Technology |
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Principal Investigator |
TERAMOTO Masaaki Kyoto Inst.Technol.Engineering and Design Professor, 工芸学部, 教授 (60026086)
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| Co-Investigator(Kenkyū-buntansha) |
KURONO Yoshiaki Amano Pharmaceutical Co.Ltd.R&D Dept.1 General Manager, 開発第一部, 部長
MATSUYAMA Hideto Kyoto Inst.Technol.Engineering and Design Assistant, 工芸学部, 助手 (50181798)
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| Project Period (FY) |
1992 – 1993
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| Keywords | Glucose Dehydrogenase / Membrane Enzyme Reactor / Water-soluble polymer / Stabilization of enzyme / NADH / NAD^+ / Plasma-graft Polymerization / Charged Membrane |
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| Research Abstract |
1. Stabilization of enzymes and accelerating effect of water-soluble polymers on enzymic reaction
(1) By addition of water-soluble cationic polymers such as polyethyleneimine(PEI) and DEAE-dextran, thermal stability of glucose dehydrogenase (GDH) was remarkably increased. It was deduced that adsorption of polymers to GDH prevents unfolding of GDH by multi-point binding.
(2) The rate of GDH-catalyzed reaction was enhanced by addition of PEI at low coenzyme(NAD()^+) concentration. The results of kinetic study suggested that the accelerating effect is due to PEI-mediated enrichment of coenzyme NAD(P)^+ near GDH interacting with PEI.
(3) In the reduction of phenylpyruvic acid(PhP) by lactate degydrogenase(LDH)/GDH conjugate system, addition of PEI was very effective for increasing reaction rate.
(4) The apparent equilibrium constant of alcohol dehydrogenase(ADH)-catalyzed reaction was increased by addition of PEI, resulting in improvement of the conversion of PhP in LDH/ADH-catalyzed reaction.
2. Effect of water-soluble polymers on performance of enzyme membrane reactor
In the continuous reduction of PhP by the LDH/GDH conjugate enzyme system using a continuoustype hollow fiber enzyme reactor, addition of PEI was effective for increasing the conversion of PhP.
3. Preparation of microporous charged membrane by plasma-graft polymerization for membrane enzyme reactor.
Microporpus charged membranes were prepared by grafting acrylic acid to microporous polyethylene membranes, and the effect of preparation condition of the membrane properties was studied. Membranes with high charge density were successfully prepared.
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