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Elucidation of Mechanism of Enzymatic Reaction in Biphasic System and Study of Chemical Reaction Engineering on Characteristics of Membrane Bioreactor

双相体系酶促反应机理的阐明及膜生物反应器特性的化学反应工程研究

基本信息

批准号：
02650707
负责人：
TERAMOTO Masaaki
金额：
$ 1.02万
依托单位：
Kyoto Institute of Technology
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

项目摘要

In order to elucidate the enzymatic reaction mechanism for hydrophobic substrates, the hydrolysis rates for several substrates were measured in biphasic and microemulsion systems. We have also carried out preliminary experiments to investigate the characteristic behavior of membrane bioreactor. The following results were obtained.1. We measured the rate of hydrolysis of the homologs of 2-naphthyl ester by using a Lewis-cell to elucidate the mechanism of the lipase-catalyzed hydrolysis in biphasic systems. On the basis of the two-film model, it was found that the hydrolysis of these substrates proceeds at the interface. The interfacial reaction rate could be correlated by the Michaelis-Menten mechanism. The values of the rate constant and the Michaelis constant were almost independent of the kinds of 2-naphthyl ester and were much greater than those for the hydrolysis in the aqueos phase.2. The enzymatic reaction rates were measured in microemulsion, which is a suitable model system to … More elucidate the interfacial reaction mechanism.(1)The hydrolysis rate of a hydrophilic substrate, acetylsalicylic acid, with alpha -chymotrypsin and lipase were measured in microemulsion. The reaction catalysed by hydrophobic lipase proceeds near the interface of microemulsion. On the other hand, the reaction by hydrophilic a -chymotrypsin proceeds in the water-pool of microemulsion, and the rate constant increased with the increase in water contents of microemulsion.(2)The hydrolysis rate of a hydrophobic substrate, 2-naphthy lacetate were also measured in microemulsion. Super-active behavior was observed in the reaction by lipase. On the other hand, the reaction for alpha -chymotrypsin was found to proceed in the water-pool of microemulsion and the rate constant increased with the increase in water contents.3. We carried out the preliminary experiments to elucidate the characteristic behavior of a membrane bioreactor for enzymatic reactions and also to examine the possibility of increasing the reaction yield of the hydrophobic substrate. flowever, the reaction yields could not be enhanced in these reaction systems. Therefore, experiments with the other hydrophobic substrates will be required in the further study. Less

为了阐明疏水底物的酶反应机理，测定了几种底物在两相和微乳液体系中的水解率。并对膜生物反应器的特性进行了初步的实验研究。取得了以下研究成果：1.我们用Lewis小室测定了2-萘酯同系物的水解率，以阐明脂肪酶在两相体系中催化水解的机理。在双膜模型的基础上，发现这些底物的水解是在界面上进行的。界面反应速率可用Michaelis-Menten机理关联。反应速率常数和米氏常数几乎与2-萘酯的种类无关，远大于水相中的水解值。在微乳液中测定了酶反应速率，这是一种适合于…的模型体系进一步阐明了界面反应机理。(1)在微乳液中测定了亲水性底物乙酰水杨酸与α-糜蛋白酶和脂肪酶的水解率。疏水脂肪酶催化的反应在微乳液界面附近进行。另一方面，亲水性的α-糜蛋白酶在微乳液的水池中进行反应，反应速率常数随着微乳液中水分含量的增加而增大。(2)测定了疏水底物2-环烷基乙酸酯在微乳液中的水解率。脂肪酶在反应中表现出超活性行为。另一方面，α-糜蛋白酶的反应是在微乳液的水池中进行的，且反应速率常数随水含量的增加而增大。我们进行了初步的实验，以阐明膜生物反应器用于酶反应的特征行为，并考察提高疏水底物反应产率的可能性。此外，在这些反应体系中不能提高反应产率。因此，在进一步的研究中，还需要对其他疏水底物进行实验。较少