Stoichiometrically Controlled Immobilization of Multiple Enzymes on Magnetic Nanoparticles by the Magnetosome Display System for Efficient Cellulose Hydrolysis
Stoichiometrically Controlled Immobilization of Multiple Enzymes on Magnetic Nanoparticles by the Magnetosome Display System for Efficient Cellulose Hydrolysis
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DOI：
10.1021/acs.biomac.5b01174
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发表时间：
2015-12-01
影响因子：
6.2
通讯作者：
Yoshino, Tomoko
中科院分区：
文献类型：
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作者：
Honda, Toru;Tanaka, Tsuyoshi;Yoshino, Tomoko
多种纤维素酶复合体的固定化因其在纤维素酶的高效水解中的应用而备受关注。本研究利用磁小体展示系统在磁性纳米颗粒(MNPs)上制备了模拟天然多重纤维素酶复合体(纤维素体)的体系。最初，绿色荧光蛋白和mCherry两种荧光蛋白被固定在MNPs上。荧光分析显示，两种不同的蛋白质在MNPs上非常接近。酶联免疫吸附分析表明，化学计量比等量的蛋白质被固定在MNPs上。其次，将内切葡聚糖酶(EG)和β-葡萄糖苷酶(BG)固定在MNPs上，制得EG/BG-MNPs。将所得的MNPs应用于纤维素酶的水解，观察到羧甲基纤维素酶的快速水解。此外，纤维素结合区与EG/BG-MNPs的融合促进了对不溶性纤维素的水解活性的提高。因此，我们可以得出结论，磁小体展示系统可以扩大在MNPs上模拟天然纤维素体组织的可能性。
The immobilization of multiple cellulase complexes receiving attention for use in the efficient hydrolysis of celluloses. In this study, the magnetosome display system was employed for the preparation of systems mimicking natural multiple cellulase complexes (cellulosomes) on magnetic nanoparticles (MNPs). Initially, two fluorescent proteins, namely, green fluorescent protein and mCherry, were immobilized on MNPs. Fluorescence analysis revealed the close proximity of two different proteins on the MNPs. Enzyme-linked immunosorbent assay analysis showed that stoichiometrically equivalent amounts of the proteins were immobilized on the MNPs. Next, endoglucanase (EG) and beta-glucosidase (BG) were immobilized on MNPs to give EG/BG-MNPs. The resulting MNPs were applied for the hydrolysis of celluloses, with rapid hydrolysis of carboxymethyl cellulose being observed. Furthermore, the fusion of the cellulose-binding domain to EG/BG-MNPs promoted improved hydrolysis activity against the insoluble cellulose. We could therefore conclude that the magnetosome display system can expand the possibilities of mimicking natural cellulosome organization on MNPs.