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β-半乳糖苷酶固定在聚偏氟乙烯膜上催化乳糖转化为半乳糖寡糖的动力学。

Kinetics of lactose conversion to galacto-oligosaccharides by β-galactosidase immobilized on PVDF membrane.

机构信息

Department of Chemical Engineering, Indian Institute of Technology Kanpur, Kanpur 208016, India.

出版信息

J Biosci Bioeng. 2013 Jun;115(6):668-73. doi: 10.1016/j.jbiosc.2012.12.014. Epub 2013 Jan 18.

Abstract

Experimental studies were made for immobilization of enzymes on microporous polyvinylidene fluoride (PVDF) membrane in order to carry out enzymatic reaction of lactose into galacto-oligosaccharides using β-galactosidase. The present work, however, is the first part in the direction of enzymatic membrane reactor studies for carrying out reaction followed by membrane based separation to purify galacto-oligosaccharides out of reaction mixture. The middle of the three compartment cell, separated by two immobilized (enzyme) membranes, was utilized to feed lactose solution; whereas, adjacent compartments were filled with distilled water. The reacted mixture solution was analyzed for tri-, tetra- and penta-forms of GOS. The formation of product GOS strongly depended on varying amounts of initial lactose concentration (ILC). Total GOS formation increased from 7% to 28% for ILC from 50 to 200 g/L. However, tri-saccharide was the major (67%) in comparison to tetra (27%) and penta (6%) forms of GOS. Further, based on Michaelis-Menten kinetics, a six-step-eleven-parameter model was developed. The model incorporated enzyme inhibition and formation of glucose and galactose separately. Simulated results from developed model matched exceeding well with experimental results.

摘要

进行了将酶固定在微孔聚偏二氟乙烯 (PVDF) 膜上的实验研究,以便使用β-半乳糖苷酶将乳糖转化为半乳糖寡糖。然而,本工作是进行酶膜反应器研究的第一部分,旨在进行反应,然后通过膜分离从反应混合物中纯化半乳糖寡糖。三隔室电池的中间部分由两个固定化(酶)膜隔开,用于进料乳糖溶液;而相邻的隔室则装满蒸馏水。对反应混合物溶液进行了三、四和五聚体形式的 GOS 的分析。产物 GOS 的形成强烈依赖于初始乳糖浓度 (ILC) 的变化量。对于 ILC 从 50 至 200 g/L 的变化,总 GOS 形成从 7%增加到 28%。然而,与四聚体(27%)和五聚体(6%)形式的 GOS 相比,三聚体是主要的(67%)。此外,基于米氏动力学,开发了一个六步十一参数模型。该模型分别纳入了酶抑制和葡萄糖和半乳糖的形成。开发模型的模拟结果与实验结果非常吻合。

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