Department of Chemical Engineering, National Chung Hsing University, 250 Kuo-Kuang Road, Taichung, 402, Taiwan.
J Biosci Bioeng. 2012 Feb;113(2):166-72. doi: 10.1016/j.jbiosc.2011.09.023. Epub 2011 Nov 8.
In this study, three membranes: regenerated cellulose (RC), glass fiber (GF) and polyvinylidene fluoride (PVDF), were grafted with 1,4-diaminobutane (DA) and activated with glutaraldehyde (GA) for lipase covalent immobilization. The efficiencies of lipases immobilized on these membranes with different hydrophobic/hydrophilic properties were compared. The lipase immobilized on hydrophobic PVDF-DA-GA membrane exhibited more than an 11-fold increase in activity compared to its immobilization on a hydrophilic RC-DA-GA membrane. The relationship between surface hydrophobicity and immobilized efficiencies was investigated using hydrophobic/hydrophilic GF membranes which were prepared by grafting a different ratio of n-butylamine/1,4-diaminobutane (BA/DA). The immobilized lipase activity on the GF membrane increased with the increased BA/DA ratio. This means that lipase activity was exhibited more on the hydrophobic surface. Moreover, the modified PVDF-DA membrane was grafted with GA, epichlorohydrin (EPI) and cyanuric chloride (CC), respectively. The lipase immobilized on the PVDF-DA-EPI membrane displayed the highest specific activity compared to other membranes. This immobilized lipase exhibited more significant stability on pH, thermal, reuse, and storage than did the free enzyme. The results exhibited that the EPI modified PVDF is a promising support for lipase immobilization.
在这项研究中,三种膜:再生纤维素(RC)、玻璃纤维(GF)和聚偏二氟乙烯(PVDF),用 1,4-二氨基丁烷(DA)接枝并用戊二醛(GA)活化,用于脂肪酶的共价固定化。比较了这些具有不同疏水性/亲水性的膜上固定化脂肪酶的效率。与在亲水性 RC-DA-GA 膜上固定化相比,固定在疏水性 PVDF-DA-GA 膜上的脂肪酶的活性增加了 11 倍以上。使用不同正丁胺/1,4-二氨基丁烷(BA/DA)接枝比的疏水性/亲水性 GF 膜研究了表面疏水性与固定化效率之间的关系。接枝 BA/DA 比增加,固定化脂肪酶的活性增加。这意味着脂肪酶活性更多地显示在疏水性表面上。此外,分别将改性的 PVDF-DA 膜接枝到 GA、表氯醇(EPI)和三聚氰胺氯(CC)上。与其他膜相比,固定在 PVDF-DA-EPI 膜上的脂肪酶显示出最高的比活性。与游离酶相比,固定化脂肪酶在 pH、热、重复使用和储存方面表现出更高的稳定性。结果表明,EPI 改性的 PVDF 是脂肪酶固定化的一种很有前途的载体。