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基于电场的多膜耦联等电聚焦酶生物反应器:以脲酶为模型的理论建模与实验验证

Isoelectrically trapped enzymatic bioreactors in a multimembrane cell coupled to an electric field: theoretical modeling and experimental validation with urease.

机构信息

University of Verona, Faculty of Science, Strada Le Grazie, Cà Vignal, 37100, Verona Italy.

出版信息

Biotechnol Bioeng. 1997 Jan 5;53(1):110-9. doi: 10.1002/(SICI)1097-0290(19970105)53:1<110::AID-BIT14>3.0.CO;2-S.

Abstract

A novel type of immobilized enzyme reactor operating under an electric field is here reported: a multicompartment immobilized enzyme reactor (MIER). In this experimental set-up, the enzyme and zwitterionic buffering ions are trapped in between two isoelectric membranes, having isoelectric point (pl) values so far apart as to trap the enzyme by an isoelectric mechanism, while allowing operation at pH optima, even when the latter pH value is quite removed from the enzyme pl. As an example, urease (pl 4.9) is trapped between a pl 4.0 and a pl 8.0 membranes, thus permitting operation (via suitable amphoteric ions buffering at pH 7.5) at the pH of optimum of activity (pH 7.5). The charged product (ammonium ions) quickly leaves the enzyme chamber under the influence of the electric field, thus allowing sustained activity for much longer time periods than in conventional reactors. As an example, while in a batch reactor 90% of original enzyme activity is lost in 200 min, only 2% activity is lost in the same period in the MIER reactor. As an additional bonus, the MIER reactor allows conversion rates of approximately 95% in a wide range of substrate concentrations, whereas batch-type reactors rarely achieve better than 50% conversion under comparable experimental conditions. (c) 1997 John Wiley & Sons, Inc.

摘要

一种新型的电场作用下的固定化酶反应器

多隔室固定化酶反应器(MIER)。在这种实验装置中,酶和两性离子缓冲离子被捕获在两个等电膜之间,这两个等电膜的等电点(pl)相差很大,以至于通过等电机制捕获酶,同时允许在最佳 pH 值下运行,即使后者 pH 值与酶的 pl 值相差很大。例如,脲酶(pl 4.9)被捕获在 pl 4.0 和 pl 8.0 的膜之间,从而允许在最佳活性 pH 值(pH 7.5)下通过合适的两性离子缓冲来进行操作(pH 7.5)。带电荷的产物(铵离子)在电场的影响下迅速离开酶室,从而使酶能够更长时间地保持活性,比传统反应器的时间长得多。例如,在分批式反应器中,90%的原始酶活性在 200 分钟内丧失,而在 MIER 反应器中,在相同的时间内仅丧失 2%的活性。作为额外的好处,MIER 反应器允许在广泛的底物浓度范围内达到约 95%的转化率,而分批式反应器在类似的实验条件下很少能达到 50%以上的转化率。(c)1997 年 John Wiley & Sons, Inc.

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